Targeting Intracellular Calcium Stores Alleviates Neurological Morbidities in a DFP-Based Rat Model of Gulf War Illness

Gulf War Illness (GWI) is a chronic multi-symptom disorder afflicting the veterans of the First Gulf War, and includes neurological symptoms characterized by depression and memory deficits. Chronic exposure to organophosphates (OP) is considered a leading cause for GWI, yet its pathobiology is not fully understood. We recently observed chronic elevations in neuronal Ca2+ levels ([Ca2+]i) in an OP- diisopropyl fluorophosphate (DFP) based rat model for GWI. This study was aimed at identifying mechanisms underlying elevated [Ca2+]i in this DFP model and investigating whether their therapeutic targeting could improve GWI-like neurological morbidities. Male Sprague-Dawley rats (9-wks) were exposed to DFP (0.5 mg/kg, s.c, 1x-daily for 5-d) and at 3-mos post DFP exposure, behavior was assessed and rats were euthanized for protein estimations and ratiometric Fura-2 [Ca2+]i estimations in acutely dissociated hippocampal neurons. In DFP rats, a sustained elevation in intracellular Ca2+ levels occurred, and pharmacological blockade of Ca2+-induced Ca2+-release mechanisms significantly lowered elevated [Ca2+]i in DFP neurons. Significant reductions in the protein levels of the ryanodine receptor (RyR) stabilizing protein Calstabin2 were also noted. Such a post-translational modification would render RyR leaky resulting in sustained DFP [Ca2+]i elevations. Antagonism of RyR with levetiracetam significantly lower elevated [Ca2+]i in DFP neurons and improved GWI-like behavioral symptoms. Since Ca2+ is a major second messenger molecule, such chronic increases in its levels could underlie pathological synaptic plasticity that expresses itself as GWI morbidities. Our studies show that treatment with drugs targeted at blocking intracellular Ca2+ release could be effective therapies for GWI neurological morbidities

Moving out of the Margins: Mattering and the International Student Experience

Deeply personal challenges confront all new students when they arrive at their college or university. Some of their core questions include: “Are we part of things; do we belong; are we central or marginal? Do we make a difference; do others care about us and make us feel we matter?” (Schlossberg, 1989, p. 6). For international students who may be marginalized by race, ethnicity, nationality, socioeconomic background or language, these questions can be particularly painful. This article recommends a new foundation for working with international students: mattering. It provides a theoretical background for the constructs of marginalization and mattering, examines their relevancy in the context of the experiences of international students, and begins to define strategies for colleges and universities to make international students feel that they do indeed matter

Repeated low-dose organophosphate DFP exposure leads to the development of depression and cognitive impairment in a rat model of Gulf War Illness

Approximately 175,000 to 250,000 of the returning veterans from the 1991 Persian Gulf War exhibit chronic multi-symptom illnesses that includes neurologic co-morbidities such as depression, anxiety and cognitive impairments. Amongst a host of causative factors, exposure to low levels of the nerve agent Sarin has been strongly implicated for expression of Gulf War Illness (GWI). Nerve agents similar to pesticides are organophosphate (OP) compounds. There is evidence from civilian population that exposure to OPs such as in agricultural workers and nerve agents such as the survivors and first-responders of the Tokyo subway Sarin gas attack suffer from chronic neurological problems similar to GWI symptoms. Given this unique chemical profile, OPs are ideal to study the effects of nerve agents and develop models of GWI in civilian laboratories. In this study, we used repeated low-dose exposure to OP agent diisopropyl fluorophosphate (DFP) over a 5-day period to approximate the duration and level of Sarin exposure during the Persian Gulf War. We tested the rats at 3-months post DFP exposure. Using a battery of behavioral assays, we observed the presence of symptoms of chronic depression, anxiety and memory problems as characterized by increased immobility time in the Forced Swim Test, anhedonia in the Sucrose Preference Test, anxiety in the Elevated Plus Maze, and spatial memory impairments in the Object Location Test, respectively. Chronic low dose DFP exposure was also associated with hippocampal neuronal damage as characterized by the presence of Fluoro-Jade staining. Given that OP exposure is considered a leading cause of GWI related morbidities, this animal model will be ideally suited to study underlying molecular mechanisms for the expression of GWI neurological symptoms and identify drugs for the effective treatment of GWIs

The effects of hypothermia on status epilepticus-induced acquired epilepsy

Status epilepticus (SE) is a type of neurological injury characterized by continuous seizure activity and can lead to molecular and pathophysiological alterations leading to plasticity changes. SE can lead to the development of AE by the process of epileptogenesis, which is a phenomenon that describes the transformation of normal brain tissue into a hyperexcitable neuronal population. It has been demonstrated both in vivo and in vitro that calcium (Ca2+) dynamics are severely altered during and after SE, and these changes play a major role in the progression of epileptogenesis. It has also been reported that preventing the rise in intracellular Ca2+ ([Ca2+]i) immediately following injury (the Ca2+ plateau) prevents the plasticity changes and ultimate development of epilepsy. Currently, there are no treatments available that can be administered following an injury to prevent the development of AE. Therefore it is clinically important to develop a therapy that can be administered after an injury to block epileptogenesis. Hypothermia is a potential therapeutic intervention. Hypothermia is used clinically to provide neuroprotection following various neurological insults such as stroke and traumatic brain injury (TBI). However, no studies have been performed to evaluate the therapeutic potential of hypothermia following SE. Hypothermia provides protection via multiple mechanisms, one of which includes modulating excitotoxic neurotransmission. It is believed to reduce Ca2+ influx by reducing NMDA receptor activation. It is unclear how hypothermia affects Ca2+ through other modes of entry. This dissertation evaluates the effects of hypothermia on the Ca2+ plateau and demonstrates the novel finding that hypothermia induced post-SE blocks the development of the Ca2+ plateau and reduces the development of AE

An Exploratory Study of Suboxone (Buprenorphine/ Naloxone) Film Splitting: Cutting Methods, Content Uniformity, and Stability

Suboxone films are U.S. Food and Drug Administration approved to treat opioid dependence. While the package insert states that films should not be cut, physicians often prescribe film fractions for treatment and tapering. There is no data to support this practice, and this study was initiated to evaluate cutting methods, content uniformity, and stability of split films. Suboxone 8-mg buprenorphine/2-mg naloxone films were split using four methods: 1) ruler/razor cut, 2) scissor cut, 3) fold/rip, and 4) fold/scissor cut. United States Pharmacopeia Chapter \u3c905\u3e was used to evaluate the weight variation and content uniformity of split films. The stability of split films stored in polybags was evaluated over 7 days. A stability-indicating high-performance liquid chromatography method was used for content uniformity and stability evaluation. The weight variation results were acceptable for the half films from all four cutting methods, but this was not true for the quarter films. The method of ruler/razor cut was determined most favorable and used for the content uniformity test. Based on the high-performance liquid chromatography results, the half films from the ruler/razor cut method met the passing criteria of United States Pharmacopeia Chapter \u3c905\u3e with acceptance values of 9.8 to 10.4 for buprenorphine and 8.4 to 11.5 for naloxone (≤15 is considered passing). The stability results indicated that both actives retained \u3e97.7% of initial strength. Four cutting methods were found to be acceptable for splitting Suboxone films into half but not quarter fractions. The half films from the ruler/razor cut method also passed United States Pharmacopeia Chapter \u3c905\u3e content uniformity test. Both actives remained stable for 7 days when the half films were stored in polybags at room temperature

The effect of group size on vigilance in a semi-solitary, fossorial marsupial (Lasiorhinus latifrons)

Prey species that congregate gain protection against predatory attacks and this advantage is often reflected by a reduction in vigilance behaviour by individuals in larger groups. Comparatively few studies have investigated vigilance in solitary animals, but those that have, found that vigilance increases as group size increases because of the threat posed by conspecifics and/or competition for resources. The southern hairy nosed wombat (Lasiorhinus latifrons) is a large fossorial, nocturnal marsupial that is neither strictly solitary nor gregarious, sharing warren systems with multiple conspecifics. We investigated the effects of conspecific presence on vigilance behaviour in this semi-solitary species. We observed wild-born, adult L. latifrons wombats in three group sizes, (Large (1♂, 3♀), Medium (1♂, 2♀) and Small (1♂, 1♀) in a captive, naturalistic environment that allowed above-ground and den behaviour monitoring. Vigilance behaviours were performed less frequently by wombats in large groups (e.g. scanning, counts/day, Large: 55, Medium: 69, Small: 115, P = 0.002) and more frequently as the distance from their nearest conspecific increased (r64 = 0.30, P = 0.016). Vigilance within burrows was also affected by social influences, with solitary wombats significantly more vigilant than those denning with a conspecific (e.g. scanning: conspecific absent: 0.13 / 5 min, present: 0.03/ 5 min, P < 0.0001). It is concluded that the presence of conspecifics reduces vigilance in L. latifrons wombats, even within burrows, and this may partially explain the occurrence of warren sharing in the wild

Survey Protocols to Investigate the Information Habits and Needs of Engineering and Engineering Technology Students and Practicing Engineers

This .pdf document contains two survey protocols created in 2016 to study the information literacy experiences and needs of undergraduate engineering and technology students and practicing engineers

The use of primary dermal fibroblast cultures to evaluate type I collagen expression in the oim model mouse

Abstract only availableOsteogenesis imperfecta type III is a heritable disorder leading to impaired connective tissue function in type I collagen containing tissues including bone fragility, blue-grey sclera, short stature, and hearing loss. Normal type I collagen is a heterotrimeric molecule containing two pro1(I) collagen chains and a similar but genetically distinct pro2(I) collagen chain. The osteogenesis imperfecta murine (oim) model mouse produces only homotrimeric type I collagen due to a single nucleotide deletion in the COL1A2 gene resulting in a non-functional pro2(I) collagen chain. The result is expression of an abnormal type I collagen molecule which leads to the above phenotype. This study is aimed at developing a methodology whereby dermal fibroblast cultures can be utilized for a variety of assays, including type I collagen RNA and protein quantification. For genotype identification, primers flanking the site of the single nucleotide deletion allow for differentiation of wild type, heterozygous and homozygous animals at the genomic level. Upon confirmation, skin was removed from both oim and wildtype mice and the dermal layer harvested. Fibroblasts originating from the dermal layer of each genotype were then cultured and grown to confluence as separate cultures and the RNA and/or protein harvested from both cell types. Total RNA was harvested using the Qiagen RNeasy kit and used to make cDNA, which was then used in conjunction with specific PCR primers to differentiate between wildtype and oim transcripts. For protein studies, the cells were treated with ascorbic acid to maximize the production of collagen prior to harvesting. Type I collagen expression was then confirmed via a western blot using a type I collagen-specific antibody. Initial results indicate the presence of both pro1(I) and pro2(I) collagen chains from harvested wildtype dermal fibroblasts, while media harvested from oim dermal fibroblasts indicated the presence of only the pro1(I) collagen chains. These results confirm the in vivo protein expression profile seen in skin of both oim and wildtype mice is exhibited in vitro in the respective dermal fibroblast cultures. These results will allow us to quantitate pro1(I) and pro2(I) collagen mRNA and protein expression levels. Future experiments will include the quantitation of type I collagen mRNA levels using RT-PCR, as well as the use of densitometry to quantitate type I collagen protein levels by western blot analysis. This in turn promises to provide insight into the mechanism of formation of abnormal type I collagen in the oim model mouse.Life Sciences Undergraduate Research Opportunity Progra

Chronic behavioral and cognitive deficits in a rat survival model of paraoxon toxicity

Organophosphate (OP) compounds, including paraoxon (POX), are similar to nerve agents such as sarin. There is a growing concern that OP agents could be weaponized to cause mass civilian causalities. We have developed a rodent survival model of POX toxicity that is being used to evaluate chronic morbidity and to screen for medical countermeasures against severe OP exposure. It is well known that the survivors of nerve gas and chronic OP exposure exhibit neurobehavioral deficits such as mood changes, depression, and memory impairments. In this study we investigated whether animals surviving severe POX exposure exhibited long-term neurological impairments. POX exposure produced overt signs of cholinergic toxicity. Rats were rescued using an optimized atropine, 2-PAM and diazepam therapy. Surviving rats were studied using established behavioral assays for identifying symptoms of depression and memory impairment 3-months after POX exposure. In the forced swim test, POX rats exhibited increased immobility time indicative of a despair-like state. In the sucrose preference test, POX rats consumed significantly less sucrose water indicating anhedonia-like condition. POX rats also displayed increased anxiety as characterized by significantly lower performance in the open arm of the elevated plus maze. Further, when tested with a novel object recognition paradigm, POX rats exhibited a negative discrimination ratio indicative of impaired recognition memory. The results indicate that this model of survival from severe POX exposure can be employed to study some of the molecular bases for OP-induced chronic behavioral and cognitive comorbidities and develop therapies for their treatment

How do professional engineers use information compared to undergradutes, and how can libraries prepare students and support engineers for future success?

Librarians at Purdue University seek to develop engineering graduates who are effective information users. Similarly, information specialists at Caterpillar Inc. are concerned with how well new hires are prepared for the information landscape at work as practicing engineers. Librarians from Purdue University and Caterpillar partnered to create and disseminate a survey to compare how students and practicing engineers seek and use information in the research process. Within a framework that asked survey participants to think about information use in a recently completed project, responses highlighted several gaps in information literacy training, including the use of external standards and internal document storage systems, documentation practices, and resource awareness. We also explored information habits and frustrations of both user populations and found that, while students have more confidence in their abilities, they consult many fewer types of literature and utilize fewer strategies for organizing information effectively. Additionally, the findings suggest students tend to use more social media tools to keep abreast of developments in their field than practicing engineers, while engineers rely heavily on internal knowledge management systems to track information generated by the company that augments externally produced information. The goal of this project was to better understand both consistencies and gaps in university engineering education and industry expectations. This paper presents preliminary findings from this project and discusses how the authors plan to use the results to update undergraduate curricula and improve library services and resources for both populations