HIV-1 enters the CNS soon after initial systemic infection. HIV-1 can induce a wide range of neurological deficits, collectively known as HIV-1-associated neurocognitive disorders (HAND). Mature neurons are not infected by HIV-1; instead, infected and/or activated glial cells release various viral and cellular factors that induce direct and/or indirect neuronal toxicity, leading to HAND. Injection drug abuse is a significant risk factor for HIV-infection, and opiate drug abusers show increased HIV-neuropathology, even with anti-retroviral treatments. Our previous work has largely modeled HIV-neuropathology using the individual viral proteins Tat or gp120, with murine striatal neurons as targets. To model disease processes more closely, the current study uses supernatant from HIV-1-infected cells. Supernatant from HIV-1SF162 (R5-tropic)-infected differentiated-U937 cells (HIV+sup) was collected and p24 level was measured by ELISA to assess the infection. We assessed HIV+sup effects on neuronal survival and neurite growth/pruning with or without concurrent exposure to morphine, an opiate that preferentially acts through µ-opioid receptors. Effects of HIV+sup ± morphine were assessed on neuronal populations, and also by time-lapse imaging of individual cells. HIV+sup caused dose-dependent toxicity over a range of p24 levels (10-500 pg/ml). Significant interactions occurred with morphine at lower p24 levels (10 and 25 pg/ml). In the presence of glia, selective neurotoxic measures were significantly enhanced and interactions with morphine were also augmented. Importantly, the arrest of neurite growth that occurred with exposure to HIV+sup was reversible unless neurons were continuously exposed to morphine. Thus, while reducing HIV-infection levels may be protective, ongoing exposure to opiates may limit recovery. During early stage of HIV-infection R5-tropic viruses are predominant, but during later stages of disease X4-tropic viruses are more predominant; co-receptor usage switch from CCR5 to CXCR4 is crucial in disease progression to AIDS. Some previous studies have shown that drugs of abuse interact with virus or viral proteins in strain/tropism-dependent manner. Therefore, we also assessed neurotoxic effects and interactions with opiates by supernatant from HIV-1LAI (X4-tropic)-infected H9 cells. Neurotoxic effects and the interactions with opiates of HIV-1LAI-supernatant are quantitatively similar to that of HIV-1SF162. Surprisingly, the cytokine/chemokine release profile of HIV-1LAI-infected H9 cells is similar to that of HIV-1SF162-infected U937 cells. Only in the presence of glia, HIV-1LAI virion induced neurotoxic effects, but no interactions with morphine were seen. Also our studies have shown that HIV-1LAI virions are slightly more neurotoxic than HIV-1SF162. Altogether, largely our results suggest that HIV+sup mediated neurotoxicity and the interactions with opiates are majorly attributed to cytotoxic factors released from infected and activated cells instead of viral strain specific factors. Although there is a correlation between opiate drug abuse and progression of HAND, the mechanisms that underlie interactions between HIV-1 and opiates remain obscure. Previous studies have shown that HIV-1 induces neurotoxic effects through abnormal activation of GSK3β. Interestingly, expression of GSK3β has shown to be elevated in the brains of young opiate abusers suggesting that GSK3β is also linked to neuropathology seen with opiate abusing patients. Thus, we hypothesized that GSK3β activation is a point of convergence for HIV- and opiate-mediated interactive neurotoxic effects. Cultures of striatal neurons were treated with HIV+sup (R5-tropic), in the presence or absence of morphine and GSK3β inhibitors. Our results show that multiple GSK3β inhibitors significantly reduce HIV-1-mediated neurotoxic outcomes, and also negate interactions with morphine that result in cell death. This suggests that GSK3β-activation is an important point of convergence and a potential therapeutic target for HIV- and opiate-mediated neurocognitive deficits.

In vitro and in situ experiments were conducted to evaluate the hypothesis that the nonclassical opioid peptide nociceptin acting on sympathetic preganglionic neurons (SPNs) inhibits spinal sympathetic outflow. First, whole cell patch recordings were made from antidromically identified SPNs from immature (12-16 day old) rat spinal cord slices. Nociceptin (0.1, 0.3, and 1 μM) concentration dependently suppressed the excitatory postsynaptic potentials (EPSPs) evoked by focal stimulation and hyperpolarized a population of SPNs; these effects were naloxone insensitive. L-Glutamate-induced depolarizations were not significantly changed by nociceptin. Results from this series of experiments indicate that nociceptin inhibits the activity of SPNs by either a presynaptic or postsynaptic site of action, whereby the peptide reduces, respectively, the amplitude of EPSPs or the excitability of SPNs. Second, intrathecal injection of nociceptin (3, 10, and 30 nmol) to urethan-anesthetized rats dose dependently reduced the mean arterial pressure and heart rate; these effects were not prevented by prior intravenous administration of naloxone (1 mg/kg). Physiological saline given intrathecally was without appreciable effects. These results, together with earlier observations of the detection of nociceptin-immunoreactive nerve fibers and nociceptin receptor immunoreactivity in the rat intermediolateral cell column, raise the possibility that the opioid peptide, which may be released endogenously, reduces spinal sympathetic outflow by depressing the activity of SPNs.

This article examines the socio-political intricacies of the United States Opiate Crisis. By first addressing the pharmaceutical origins of Oxycontin and its pain relief benefits within the United States, I construct a framework by which a conversation about substance abuse can move forward. Within the first chapter I provide background into the arguments for medicalization against personal responsibility as it relates to opiate withdrawal and seeking treatment for the prior. Furthermore, I include subheadings to further provide insight into Medically Assisted Treatment Centers and their function on the local level. I contrast these modern treatment models with the Reagan War on Drugs mentalities and illustrate a larger societal tonal shift towards increased medicalization. My second chapter addresses the bulk of my theoretical frameworks, including spatial and feminist theories to construct an argument about patriarchal dominance in relation to factors such as homelessness, race, and socioeconomic status. Finally, in my third chapter I examine the current debate regarding whether or not the U.S. Opiate Crisis can truly be considered a crisis. I address the arguments for why it could still be considered a widespread crisis, and end on the central argument for the crisis being a symptom of Disease of Despair.

No description available.

The opioid epidemic is an issue that has ravaged much of the United States, and specifically Appalachia. Many different academic disciplines have attempted to provide a solution to no avail. This ethnographic study investigates the social nature of addiction & recovery related to problematic opiate use. Through semi-structured interviews and participant observation with clients at a drop-in mental health and addiction services center, the study explores the social conditions which facilitate problematic substance use in addition to vetted strategies to reach and maintain active recovery from opiate misuse. The conclusion focuses on how addiction and recovery are inherently social exercises that are heavily influenced by one's social network and the social contexts they live or have lived in. Using an anthropological perspective, this study shows the value that social science and an anthropological perspective in particular, can provide on such a pervasive and unsettling issue.

M5 muscarinic receptors are associated with dopamine neurons of the ventral tegmental area (VTA) and substantia nigra, and provide an important excitatory input to the mesolimbic dopamine system. Here, I studied locomotion induced by systemic morphine (3, 10, 30 mg/kg, i.p.) in M5 knockout mice of the C57Bl/6 (B6) and CD1 x 129SvJ (129) background strains. M5 knockout mice of both strains showed reduced locomotion in response to 30 mg/kg morphine, while only B6 M5 knockout mice showed reduced locomotion in response to 10 mg/kg morphine. In B6 wild-type mice VTA pre-treatment with the non subtype-selective muscarinic receptor antagonist atropine (3 mg per side), but not the non subtype-selective nicotinic receptor antagonist mecamylamine (5 mg per side), reduced locomotion in response to 30 mg/kg (i.p.) morphine to a similar extent as systemic M5 knockout, suggesting that the reduced morphine-induced locomotion in M5 knockout mice was due to the loss of M5 receptors on VTA dopamine neurons. By contrast, in M5 knockout mice, either intra-VTA atropine or mecamylamine alone increased locomotion by almost 3 times relative to saline, and potentiated morphine-induced locomotion. Therefore, in M5 knockout mice, more clearly than in wild-type mice, blockade of either VTA muscarinic or nicotinic receptors activated locomotion. Infusions of morphine (50 ng) into the VTA increased nucleus accumbens dopamine efflux in urethane-anesthetized wild-type mice. Either M5 knockout or pre-treatment with VTA scopolamine (50 ug) in wild-type mice blocked accumbal dopamine efflux in response to VTA morphine. Therefore, M5 receptors are critical for excitation of dopamine neurons by intra-VTA morphine, suggesting that the reduced locomotion produced by systemic morphine in M5 knockout mice was, in part, due to loss of M5-mediated excitation of VTA dopamine neurons by opiates. The locomotion data also show that in the absence of M5 receptors, cholinergic afferents to mesolimbic dopamine neurons are inhibitory. This supports and extends the conclusions from many studies that non-M5 muscarinic receptors inhibit, and M5 receptors excite, dopamine neurons. Loss of M5-mediated excitation results in reduced acute effects of opiates.

M5 muscarinic receptors are associated with dopamine neurons of the ventral tegmental area (VTA) and substantia nigra, and provide an important excitatory input to the mesolimbic dopamine system. Here, I studied locomotion induced by systemic morphine (3, 10, 30 mg/kg, i.p.) in M5 knockout mice of the C57Bl/6 (B6) and CD1 x 129SvJ (129) background strains. M5 knockout mice of both strains showed reduced locomotion in response to 30 mg/kg morphine, while only B6 M5 knockout mice showed reduced locomotion in response to 10 mg/kg morphine. In B6 wild-type mice VTA pre-treatment with the non subtype-selective muscarinic receptor antagonist atropine (3 mg per side), but not the non subtype-selective nicotinic receptor antagonist mecamylamine (5 mg per side), reduced locomotion in response to 30 mg/kg (i.p.) morphine to a similar extent as systemic M5 knockout, suggesting that the reduced morphine-induced locomotion in M5 knockout mice was due to the loss of M5 receptors on VTA dopamine neurons. By contrast, in M5 knockout mice, either intra-VTA atropine or mecamylamine alone increased locomotion by almost 3 times relative to saline, and potentiated morphine-induced locomotion. Therefore, in M5 knockout mice, more clearly than in wild-type mice, blockade of either VTA muscarinic or nicotinic receptors activated locomotion. Infusions of morphine (50 ng) into the VTA increased nucleus accumbens dopamine efflux in urethane-anesthetized wild-type mice. Either M5 knockout or pre-treatment with VTA scopolamine (50 ug) in wild-type mice blocked accumbal dopamine efflux in response to VTA morphine. Therefore, M5 receptors are critical for excitation of dopamine neurons by intra-VTA morphine, suggesting that the reduced locomotion produced by systemic morphine in M5 knockout mice was, in part, due to loss of M5-mediated excitation of VTA dopamine neurons by opiates. The locomotion data also show that in the absence of M5 receptors, cholinergic afferents to mesolimbic dopamine neurons are inhibitory. This supports and extends the conclusions from many studies that non-M5 muscarinic receptors inhibit, and M5 receptors excite, dopamine neurons. Loss of M5-mediated excitation results in reduced acute effects of opiates.

abstract: There has been a rise in heroin use throughout the United States due to doctors increasingly prescribing painkillers to patients with chronic pain (Kanouse & Compton, 2015; Compton, Boyle, & Wargo, 2015). Individuals get addicted to painkillers and, when their doctor will no longer prescribe them, turn to alternative methods of relief; heroin is often their cheapest option (Kolodny, Courtwright, Hwang, Kreiner, Eadie, Clark, & Alexander 2015). Heroin users are three to four times more likely to die from overdose than other types of drug users (Darke & Hall, 2003). The purpose of this study is to determine the likelihood that heroin users successfully reenter the community upon release from prison in comparison to other types of drug users. There are several re-entry outcomes that can be considered “success”; this study measures success as an index of the quality of the returning offender’s familial relationships as well as recidivism. The data used for this analysis is the Serious and Violent Offender Reentry Initiative (SVORI). The sample consists of male offenders, aged 18 years and older, who have been convicted of and imprisoned for a serious or violent crime. Findings suggest familial social support does not have an effect on heroin use, but heroin use increases the risk of recidivism. These findings will provide a context for rehabilitation of heroin offenders and will launch future research focusing on the differences between heroin users and other types of drug users. / Dissertation/Thesis / Masters Thesis Criminology and Criminal Justice 2016

Drugs of Abuse (DOAs) are among the single largest contributor to crime in the United States and present a high cost to society in terms of financial costs and physical/mental well-being of individuals. The forensic community requires a variety of validated methods to detect and analyze DOAs in a variety of different sample types, and most developed methods utilize a liquid or gas chromatography (GC or LC) separation system paired to a mass spectrometer (MS) detection detector. Capillary Electrophoresis (CE) based separation techniques have also been experimented with due to this technique’s high efficiency and speed, high resolving power, low sample consumption, and potentially lower cost when compared to GC or LC based techniques, even though the sensitivity of these systems is perceived to be weaker. The goal of this research to develop a CE-MS/MS method utilizing the ZipChipTM to demonstrate it can accurately and reliably detect and quantify DOAs. The DOAs analyzed for this method were opioids and benzodiazepines, and these were 6-monacetylmorphine, 7-aminoclonazepam, codeine, diazepam, dihydrocodeine, 2-Ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine fentanyl, heroin, hydrocodone, hydromorphone, meperidine, methadone, morphine, norfentanyl, oxycodone, and oxymorphone. Standard Practices for Method Validation in Forensic Toxicology guidelines from the Academy Standards Board (ASB) of Toxicology were used as the template for this validation; samples were prepared and analyzed as neat standards in diluent, blood and urine were assessed for interferences, ionization suppression/enhancement, and extraction recovery. The total runtime for the method was 3.5 minutes, with the retention time range being 1.4 to 2.9 minutes. All samples were prepared using compound standards diluted in metabolite diluent, which consisted of methanol, ammonium acetate, and water prior to injection. The calibration curves consisted of eight calibrator samples that ranged from 0.5 ng/ml to 200 ng/ml for all analytes, and a linear model was used for each compound. The minimum acceptable 𝑅2 value was set to >0.98, and each curve had a weighing factor of 1𝑥2. Each curve for most of the compounds achieved the minimum requirement apart from two Codeine curves (0.9781 and 0.9785) and 7-aminoclonazepam (0.9791). Bias and precision were assessed at three concentrations- 5, 100, and 150 ng/ml. The minimum requirement for bias and precision for a compound was if the percent bias or coefficient of variation was within +/- 20%. Most compounds in this method exhibit acceptable levels of bias (except for Dihydrocodeine which had a bias of 24.58% at 100 ng/ml), and the only compounds to meet the minimum requirement for precision were 6-MAM, 7-aminoclonazepam, diazepam, fentanyl, methadone, and morphine. The limit of detection and limit of quantitation were both set at the lowest calibrator level of 0.5 ng/ml, and no carryover was observed in this method. No interferences occurred due to both deuterated internal standards and from common compounds such as benzylecogine, cocaine, and lidocaine, but blood cause signal interference with fentanyl and urine caused signal interference with methadone and norfentanyl. Ionization suppression and enhancement was observed for a majority of the compounds, and this observation will need to be assessed as to the effect it has on validation parameters in the future. The results collected suggest that accurate, reliable, and sensitive data may be collected if a compound has a specifically paired deuterated internal standard included in the sample. The speed of the suggested method and the minimal sample preparation could be desirable for forensic use. Further testing will need to be conducted to fully validate this method for blood and urine.

The perinate is particularly risk-prone to chemical species which have the potential of inducing neuronal apoptosis or necrosis and thereby adversely altering development of the brain, to produce life-long functional and behavioral deficits. This paper is an overview for many substances of abuse, but the purview is much more broadened by the realization that even elevated levels of estrogens and corticosteroids in the pregnant mother can act as neuroteratogens, by passing via the placenta and altering neural development or inducing apoptosis in the perinate. Finally, therapeutic risks of anesthetics are highlighted, as these too induce neuronal apoptosis in the neonate by either blocking N-methyl-D-aspartate receptors or by acting as gamma-aminobutyric acid agonists. By understanding the mechanisms involved it may ultimately be possible to interrupt the mechanistic scheme and thereby prevent neuroteratological processes.