Exploring the Epistemology of Illicit Drugs

In this essay I explore the epistemology of drugs in America. That is, how Americans come to know and define drugs and their users; and, in turn, how those definitions manifest in social institutions. I argue that at the present moment, the cultural environment surrounding the ways in which Americans define and experience drug use— whether it is deemed as acceptable usage or a threat—is determined by three main institutions. The first and most predominant is the criminal justice system, which operates according to a regime of “narcopolitics.

Teachers’ Help-Seeking Perceptions and Workplace Psychological Safety

This objective of this study was to investigate teacher mental health, attitudes toward psychological help-seeking, and perceptions of psychological safety within schools. Findings from this study of 600 Canadian educational professionals revealed a higher frequency and severity of distress than in the general population, warranting intervention. Attitudes toward psychological help-seeking were generally positive, though less favorable than comparable published studies. Overall, participants rated their workplaces as “somewhat” psychologically safe. However, there were low ratings of work-life balance and respectful treatment of mental illness by leadership. Implications and recommendations of these findings were discussed

Imperfect Heart

Herein lies a personal inquiry and collection of beautiful ruin in an imperfect architectural world. The journey into abandoned ruin, born out of a curiosity to see what comes of viewing architecture in its darkest hours. This fascination with abandoned architectural spaces brings forth more than a visual sense of decay, it emulates an essence of another world, charged with emotion. This human intrigue, leads to a photographical journey of exploration into ruin from different viewpoints, ultimately focusing on the connection between the individual and home as ruin becomes the symbiotic thread between the two. It is primarily within these photographs that this thesis becomes a personification of the ruined home of an imperfect individual. The personal journey is put forth in three stages. Three books portray different perspectives to exploring ruin, each one delving deeper into an onslaught of ruin in time. The ruin begins by breaking into the world of ruin through views and definitions from the perspective of an outsider in Seeing Ruin (Book 1). The journey into ruin continues in Creating Ruin (Book 2) by an outsider’s contrivance of ruin in an imaginary world of ruin, exploring such themes as age, texture, time and loss. Lastly Living Ruin (Book 3) is explored by an insider, feeling ruin and living within it. This thesis opposes the negative notion of ruin inherent in its dictionary definition. In searching for the light within ruin, this inquiry seeks a place and use for ruin within the world of architecture. This depiction strives to show the beauty of ruin, the light in darkness. It strives for a discussion of a darker side, which is normally withheld amongst its viewers. Creating art and architecture in part from failure imbues the palpable reality of life within the human household. This journey is one view of a time filled with ruin, in one person’s world

Agonist-Induced Endocytosis of Muscarinic Cholinergic Receptors: Relationship to Stimulated Phosphoinositide Turnover

The ability of muscarinic cholinergic receptors to activate phosphoinositide turnover following agonist-induced internalization has been investigated. Incubation of SH-SY5Y neuroblastoma cells with oxotremorine-M resulted in a time-dependent endocytosis of both muscarinic receptors and Α subunits of G q and G 11 , but not of isoforms of phosphoinositide-specific phospholipase C, into a subfraction of smooth endoplasmic reticulum (V 1 ). Agonist-induced increases in diacylglycerol mass and in 32 P-phosphatidate labeling, much of which was of the tetraenoic species, were also observed in the V 1 fraction, but these increases persisted when the agonist-induced translocation of receptors into the V 1 fraction was blocked. All enzymes of the phosphoinositide cycle were detectable in the V 1 fraction. However, with the exception of phosphatidylinositol 4-kinase, none was enriched when compared with cell lysates. Both 32 P-labeling studies and enzyme assays point to a very limited capacity of this fraction to synthesize phosphatidylinositol 4,5-bisphosphate, whereas the synthesis of phosphatidylinositol 4-phosphate is robust. These results indicate that endocytosed receptors do not appear to retain their ability to activate phosphoinositide turnover. The availability of the substrate for phospholipase C, phosphatidylinositol 4,5-bisphosphate, may be one factor that limits the activity of muscarinic receptors in this subcellular compartment.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65644/1/j.1471-4159.1997.68041473.x.pd

Cytoskeletal and Phosphoinositide Requirements for Muscarinic Receptor Signaling to Focal Adhesion Kinase and Paxillin

The mechanism whereby agonist occupancy of muscarinic cholinergic receptors elicits an increased tyrosine phosphorylation of focal adhesion kinase (FAK) and paxillin has been examined. Addition of oxotremorine-M to SH-SY5Y neuroblastoma cells resulted in rapid increases in the phosphorylation of FAK ( t 1/2 = 2 min) and paxillin that were independent of integrin-extracellular matrix interactions, cell attachment, and the production of phosphoinositide-derived second messengers. In contrast, the increased tyrosine phosphorylations of FAK and paxillin were inhibited by inclusion of either cytochalasin D or mevastatin, agents that disrupt the cytoskeleton. Furthermore, phosphorylation of FAK and paxillin could be prevented by addition of either wortmannin or LY-294002, under conditions in which the synthesis of phosphatidylinositol 4-phosphate was markedly attenuated. These results indicate that muscarinic receptor-mediated increases in the tyrosine phosphorylation of FAK and paxillin in SH-SY5Y neuroblastoma cells depend on both the maintenance of an actin cytoskeleton and the ability of these cells to synthesize phosphoinositides.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66183/1/j.1471-4159.1998.70030940.x.pd

Unit Activity in the Hypothalamus and Striatum of the Rat During Learning

Unit activity was recorded from the hypothalamus and striatum of 80 freely moving rats during an appetitive classical conditioning situation. Responses to auditory stimuli were observed from 118 units before and during a conditioning procedure in which presentation of food occurred one second after the onset of an auditory stimulus. A large proportion of units (111) showed changed responses to the CS during conditioning. Only 8 of these, however, showed new conditioned responses of the very shortest latency measured, 20 msec. after CS onset. These were interpreted as likely sites of rerouting of the stimulus information within the brain as a result of learning. They were located largely near the intersection of hypothalamic and striatal structures. A transient increase in rate of background firing over trials was recorded following the onset of conditioning among hypo­thalamic units, suggesting they may temporarily represent a dynamic trace of the new learning. No significant differences were found between areas studied in order of appearance over trials of the conditioned responses. However, as a group, the conditioned responses studied here, appeared significantly earlier than a group of cortical neurons studied under similar conditions. There was greater generalization of response to the CS- by units of the basal ganglia than other areas, suggesting they may be of importance in inhibition of response to the CS-

The effect of glycosidases on the survival of rat erythrocytes in circulation

Enzymic removal of sialyl groups from mammalian erythrocytes resulted in their rapid sequestration from circulation subsequent to autologous transfusion. It has been demonstrated by many investigators that the terminal [beta]--galactosyl group, exposed on red blood cell by in vitro desialosylation, is recognized by an autoimmune anti-galactosyl IgG and/or by a lectin-like receptor on monocytes and macrophages. It is demonstrated herein that the disaccharide structure [beta]--Galp-(1-->3)--GalpNAc (a) is masked in normal rat RBC, but exposed in asialo-RBC; (b) could be detected with fluorescently-labeled peanut agglutinin; (c) could be released from the asialo-RBC with an endo-N-acetyl-[alpha]--galactosaminidase; and (d) upon its removal by treatment with the endo-N-acetyl-[alpha]--galactosaminidase, enhances the survival of the asialo-RBC in circulation.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/27216/1/0000220.pd

Signal transducer and activator of transcription-5 mediates neuronal apoptosis induced by inhibition of Rac GTPase activity.

In several neuronal cell types, the small GTPase Rac is essential for survival. We have shown previously that the Rho family GTPase inhibitor Clostridium difficile toxin B (ToxB) induces apoptosis in primary rat cerebellar granule neurons (CGNs) principally via inhibition of Rac GTPase function. In the present study, incubation with ToxB activated a proapoptotic Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway, and a pan-JAK inhibitor protected CGNs from Rac inhibition. STAT1 expression was induced by ToxB; however, CGNs from STAT1 knock-out mice succumbed to ToxB-induced apoptosis as readily as wild-type CGNs. STAT3 displayed enhanced tyrosine phosphorylation following treatment with ToxB, and a reputed inhibitor of STAT3, cucurbitacin (JSI-124), reduced CGN apoptosis. Unexpectedly, JSI-124 failed to block STAT3 phosphorylation, and CGNs were not protected from ToxB by other known STAT3 inhibitors. In contrast, STAT5A tyrosine phosphorylation induced by ToxB was suppressed by JSI-124. In addition, roscovitine similarly inhibited STAT5A phosphorylation and protected CGNs from ToxB-induced apoptosis. Consistent with these results, adenoviral infection with a dominant negative STAT5 mutant, but not wild-type STAT5, significantly decreased ToxB-induced apoptosis of CGNs. Finally, chromatin immunoprecipitation with a STAT5 antibody revealed increased STAT5 binding to the promoter region of prosurvival Bcl-xL. STAT5 was recruited to the Bcl-xL promoter region in a ToxB-dependent manner, and this DNA binding preceded Bcl-xL down-regulation, suggesting transcriptional repression. These data indicate that a novel JAK/STAT5 proapoptotic pathway significantly contributes to neuronal apoptosis induced by the inhibition of Rac GTPase

A Role for the Small Molecular Weight GTPases, Rho and Cdc42, in Muscarinic Receptor Signaling to Focal Adhesion Kinase

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66289/1/j.1471-4159.2000.0742010.x.pd

Activation of muscarinic cholinergic receptors enhances the volume-sensitive efflux of myo-inositol from SH-SY5Y neuroblastoma cells

A mechanism used by cells to regulate their volume under hypo-osmotic conditions is the release of organic osmolytes, one of which is myo-inositol. The possibility that activation of phospholipase-C-linked receptors can regulate this process has been examined for SH-SY5Y neuroblastoma cells. Incubation of cells with hypo-osmolar buffers (160–250 mOsm) led to a biphasic release of inositol which persisted for up to 4 h and could be inhibited by inclusion of anion channel blockers – results which indicate the involvement of a volume-sensitive organic anion channel. Inclusion of oxotremorine-M, a muscarinic cholinergic agonist, resulted in a marked increase (80–100%) in inositol efflux under hypo-osmotic, but not isotonic, conditions. This enhanced release, which was observed under all conditions of hypo-osmolarity tested, could be prevented by inclusion of atropine. Incubation of the cells with either the calcium ionophore, ionomycin, or the phorbol ester, phorbol 12-myristate 13-acetate, partially mimicked the stimulatory effect of muscarinic receptor activation when added singly, and fully when added together. The ability of oxotremorine-M to facilitate inositol release was inhibited by removal of extracellular calcium, depletion of intracellular calcium or down-regulation of protein kinase C. These results indicate that activation of muscarinic cholinergic receptors can regulate osmolyte release in this cell line.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65241/1/j.1471-4159.2003.02021.x.pd