Isolation and characterization of Pisum sativum apyrases, PsNTP9 and PsNTP9-DM, cloned and expressed in Escherichia coli

Adenosine triphosphate (ATP) is widely known as a fuel source for many biochemical processes, and to a lesser degree also as a signaling molecule in plants and animals. When plants are subjected to biotic or abiotic stress or undergoing exocytosis, they release ATP into the extracellular matrix (ECM). The release of ATP sets off a signal transduction pathway, first rapidly increasing the concentrations of cytosolic calcium, reactive oxygen species, and nitric oxide. How these changes specifically influence physiology is the object of much research in both plants and animals. Some of the changes that are affected influence growth and development, stomatal function, and gravitropism. Apyrases and other phosphatases control the concentration of the released nucleotides by breaking phosphate bonds from nucleoside triphosphates and diphosphates. Research aimed at the discovery of receptors, signaling pathway components, and processes has been successful to some extent. There are now known purinergic receptors in both plants and animal cells. We have cloned a truncated version of Pisum sativum (ps) NTP9. We used a pET-22B vector to add a histidine tag and transformed the vector into the BL21 Escherichia coli with a T7 promoter to enable IPTG induction of the LAC operon and expression of the enzyme. The pET-22B vector was incubated in separate samples with BL21 cells. Cells were propagated, and the expression of recombinant proteins PsNTP9, and separately, a double mutant PsNTP9-DM with a second calmodulin-binding domain, were induced ectopically. Cells were broken open by shaking them and mixing them with lysis buffer. Centrifugation was performed to separate the supernatant containing the released apyrases from the particulate wall fraction. The enzymes were purified by affinity chromatography, then their purity was evaluated by sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE). Western blots were performed to verify presence of the apyrases using a commercial anti-histidine antibody to detect PsNTP9 and PsNTP9-DM. Once suitable amounts of our proteins of interest were harvested, we performed Bradford assays to determine the protein concentration of the samples and carried out an apyrase activity assay to determine the specific activity of the purified enzymes and compare it to that of other known phosphatases.Plant Biolog

Dynamics of a Monolayer of Microspheres on an Elastic Substrate

We present a model for wave propagation in a monolayer of spheres on an elastic substrate. The model, which considers sagittally polarized waves, includes: horizontal, vertical, and rotational degrees of freedom; normal and shear coupling between the spheres and substrate, as well as between adjacent spheres; and the effects of wave propagation in the elastic substrate. For a monolayer of interacting spheres, we find three contact resonances, whose frequencies are given by simple closed-form expressions. For a monolayer of isolated spheres, only two resonances are present. The contact resonances couple to surface acoustic waves in the substrate, leading to mode hybridization and "avoided crossing" phenomena. We present dispersion curves for a monolayer of silica microspheres on a silica substrate, assuming adhesive, Hertzian interactions, and compare calculations using an effective medium approximation to a discrete model of a monolayer on a rigid substrate. While the effective medium model does not account for discrete lattice effects at short wavelengths, we find that it is well suited for describing the interaction between the monolayer and substrate in the long wavelength limit. We suggest that a complete picture of the dynamics of a discrete monolayer adhered to an elastic substrate can be found using a combination of the results presented for the discrete and effective medium descriptions. This model is potentially scalable for use with both micro- and macroscale systems, and offers the prospect of experimentally extracting contact stiffnesses from measurements of acoustic dispersion

An Investigation of the Phenomenon of Separation in the Air Flow Around Simple Quadric Cylinders

The tests, conducted at the Guggenheim Aeronautical Laboratory of Stanford University, to investigate the phenomenon of separation in the air flow past geometric shapes are described in this report

Silhouettes

Silhouettes Silhouettes began as an experiment in reduction. I was looking to capture emptiness; to photograph nothing. However, it was important to work within the confines of the program of the camera. I didn’t want to cheat the apparatus, and end up with something not quite photography. I wanted to meet “straight photography” on its own terms, which got me thinking about the function of a photograph. Photography has long served as a medium for documentation. From its genesis to today, it has been used to a breadth of ends as a creator of records. One particularly interesting case is crime scene photography. Forensic photographers are given the responsibility to photograph a scene after an event has taken place, with the hope that their documentation will help shed light on the past. In a sense, they attempt to create a document of something that isn’t there. I followed that train of thought in Silhouettes. I began photographing as if I was arriving “after the fact.” I approached scenes with a curious eye, hoping my scrutiny would lead to discovery. Shortly after adopting this way of working, I came across an article by Ana Longoni titled, “Photographs and Silhouettes: Visual Politics in the Human Rights Movement of Argentina.” In the article, Longoni focused on the “aesthetic implications of the visual strategies [in the] protests and remembrances of those who disappeared under the Argentinian dictatorship of the 1970s and 80s”. Photographs became a big part of the protests and remembrances, as documents of existence. During the Argentinean dictatorship of 1976-83, 500 concentration camps were established, where approximately 30,000 people disappeared. Small posters were made with images of the disappeared. These images were worn on bodies or held up on signs as Argentineans protested around the Plaza de Mayo. “These images of the disappeared reaffirmed the existence of a biography that predated these subjects’ kidnapping, an existence that was categorically negated by the regime”. The photographs declared that this was, this took place: this person existed. “The photographic apparatus contains this temporal ambiguity of what still is and what no longer is”. I find that opportunity for temporal ambiguity, and specificity, interesting. Using the psychology of the crime scene photograph as the primary model, Silhouettes explores the function of the photograph as document, and its implications of biography. A black photograph was included in the exhibition, as part of a sound installation. Attached to the top of the frame were two binaural microphones that were plugged in to a recorder hidden behind the frame. Plugged into the recorder was a pair of headphones, hung on a nail below the frame. The viewer was invited to put the headphones on while looking at the photograph. What the viewer heard through the headphones was realtime 3D audio of the noise of the room, amplified to a level above normal hearing. One could hear every footstep, shuffle, and conversation. Behind the glass, the black image became a mirror. The viewer saw themselves and everything taking place around them. The amplified audio and alluring reflection created a heightened sense of awareness, that simultaneously connected and disconnected the viewer from the space of the gallery. They became hyper aware of the space they were in, but also felt a sense of its otherness, as their experience of it was mediated through the photograph and the headphones. For installation shots/images please visit: http://curtiswallen.com/silhouettes

The Federal Tort Claims Act - Absolute Liability, The Discretionary Function Exception, Sonic Booms; Laird v. Nelms

[A]lthough the legislative history of the FTCA lends great support for the argument that the doctrine of absolute liability is an acceptable theory to employ to seek recovery under the FTCA, the Supreme Court has chosen to rely on the Dalehite decision and completely overlook later Supreme Court interpretations. The legislative history of the FTCA also indicates that the discretionary function exception must always be confronted, regardless of the theory one proceeds under to seek recovery

The Effectiveness of Treatment Plans for Incarcerated Individuals with Personality Disorders

This literature review aims to examine the types of treatment plans incarcerated individuals diagnosed with personality disorders receive and their effectiveness. I expect to find that inmates who receive Dialectical Behavior Therapy (DBT) will have higher rates of completing treatment. DBT consists of individual and group therapy, with a focus on the person’s emotions and actions and how they are related. Research indicates that DBT is an effective treatment strategy for female inmates with borderline personality disorder, as well as for inmates with comorbid borderline and antisocial personality disorders. Substance abuse treatment strategies have also been found to be effective for inmates with antisocial personality disorder co-occurring with substance abuse disorders. Limitations of this review include that there is not sufficient research on evidence-based treatment for antisocial personality disorder alone, on the co-occurrence of ASPD with other disorders, and on the recidivism rate of individuals who receive such treatment interventions. Additional research shows that many inmates will only seek treatment to reduce their sentences or to be transferred to a hospital. More research should be conducted on the treatment of individuals diagnosed with antisocial personality disorder

Post-Traumatic Stress Disorder: A Look into the Cause, Gender Differences, and Treatment

Posttraumatic stress disorder, better known as PTSD came to light during the Vietnam Era. Throughout history, this stress disorder has been called various things in the 150 years since it was first recognized. However, each new word had several characteristics in common, such as re-experiencing, numbing and physiological arousal. The process of Darwinian “natural selection” corroborated the evolution of people with highly developed stress responses. Those pre-historic people with the most useful “fight or flight” reflexes became our ancestors. An example of this is that during the 19th Century, PTSD was called “Railway Spine” and was associated with what we would today call “hysterical” physical symptoms such as “anxiety” expressed as bodily complaints (Ursano et al. 2007). $ is disorder was seen in individuals who had been involved in railway accidents but had not suffered bodily injuries. During traumatic events, it is expected for people to be resilient. However, some endure the psychological stress of the trauma exposure in a different way, developing distress, psychiatric illness, and exhibit health risk behaviors. In fact, after trauma exposure an altered sense of safety, increased fear and arousal, and concern for the future, can affect not only those who acquire mental health troubles (Ursano et al., 2007). It may also affect those who continue to work and care for their families and loved ones (Ursano et al., 2007)

5-0 Is Not Coming To Save You: Examining The Lack Of Police Intervention In LGBTQ Intimate Partner Violence

Intimate partner violence (IPV) has garnered a lot of attention by the criminal justice system, the media and, academia. Intimate partner violence laws have seen drastic changes over the past hundred years often coupled with the changes of culture and society. Though intimate partner violence surely warrants that attention, the growing and important issue of lesbian, gay, bisexual, transgender, and queer (LGBTQ) intimate partner violence has largely been ignored. The ongoing struggle of LGBTQ community for equality has not as of yet been a catalyst for drastic legal change. Utilizing narratives from victims of intimate partner violence from around the nation, this paper hopes to prove that the police and criminal justice system have failed to provide protective measures to victims of LGBTQIPV. That this failure is not solely from personal biases of the everyday police officer but more importantly from the institutional and societal. By using a Institutional ethnography, which is rooted in the mapping of social interactions of rules of governance, these victim narratives will show the system of oppression and marginalization that creates these harsh realities for LGBTQ victims and promotes their continued victimization. The five prevailing themes, 1) Violence; 2) Lack of police response; 3) gender roles; 4) fear of outing or deportation; and 5) services provided, found in the qualitative data will be contextualized and explained. Parallels to the broader fight for equality of the LGBTQ community the potential solutions, will also be discussed