Covalent protein adduction by drugs of abuse

Recreational abuse of the drugs cocaine, methamphetamine, and morphine continues to be prevalent in the United States of America and around the world. While numerous methods of detection exist for each drug, they are generally limited by the lifetime of the parent drug and its metabolites in the body. However, the covalent modification of endogenous proteins by these drugs of abuse may act as biomarkers of exposure and allow for extension of detection windows for these drugs beyond the lifetime of parent molecules or metabolites in the free fraction. Additionally, existence of covalently bound molecules arising from drug ingestion can offer insight into downstream toxicities associated with each of these drugs. This research investigated the metabolism of cocaine, methamphetamine, and morphine in common in vitro assay systems, specifically focusing on the generation of reactive intermediates and metabolites that have the potential to form covalent protein adducts. Results demonstrated the formation of covalent adduction products between biological cysteine thiols and reactive moieties on cocaine and morphine metabolites. Rigorous mass spectrometric analysis in conjunction with in vitro metabolic activation, pharmacogenetic reaction phenotyping, and computational modeling were utilized to characterize structures and mechanisms of formation for each resultant thiol adduction product. For cocaine, data collected demonstrated the formation of adduction products from a reactive arene epoxide intermediate, designating a novel metabolic pathway for cocaine. In the case of morphine, data expanded on known adduct-forming pathways using sensitive and selective analysis techniques, following the known reactive metabolite, morphinone, and a proposed novel metabolite, morphine quinone methide. Data collected in this study describe novel metabolic events for multiple important drugs of abuse, culminating in detection methods and mechanistic descriptors useful to both medical and forensic investigators when examining the toxicology associated with cocaine, methamphetamine, and morphine.

HIV infection and drugs of abuse: role of acute phase proteins

Background HIV infection and drugs of abuse such as methamphetamine (METH), cocaine, and alcohol use have been identified as risk factors for triggering inflammation. Acute phase proteins such as C-reactive protein (CRP) and serum amyloid A (SAA) are the biomarkers of inflammation. Hence, the interactive effect of drugs of abuse with acute phase proteins in HIV-positive subjects was investigated. Methods Plasma samples were utilized from 75 subjects with METH use, cocaine use, alcohol use, and HIV-positive alone and HIV-positive METH, cocaine, and alcohol users, and age-matched control subjects. The plasma CRP and SAA levels were measured by ELISA and western blot respectively and the CD4 counts were also measured. Results Observed results indicated that the CRP and SAA levels in HIV-positive subjects who are METH, cocaine and alcohol users were significantly higher when compared with either drugs of abuse or HIV-positive alone. The CD4 counts were also dramatically reduced in HIV-positive with drugs of abuse subjects compared with only HIV-positive subjects. Conclusions These results suggest that, in HIV-positive subjects, drugs of abuse increase the levels of CRP and SAA, which may impact on the HIV infection and disease progression.

Covalent Protein Adduction by Drugs of Abuse

Recreational abuse of the drugs cocaine, methamphetamine, and morphine continues to be prevalent in the United States of America and around the world. While numerous methods of detection exist for each drug, they are generally limited by the lifetime of the parent drug and its metabolites in the body. However, the covalent modification of endogenous proteins by these drugs of abuse may act as biomarkers of exposure and allow for extension of detection windows for these drugs beyond the lifetime of parent molecules or metabolites in the free fraction. Additionally, existence of covalently bound molecules arising from drug ingestion can offer insight into downstream toxicities associated with each of these drugs. This research investigated the metabolism of cocaine, methamphetamine, and morphine in common in vitro assay systems, specifically focusing on the generation of reactive intermediates and metabolites that have the potential to form covalent protein adducts. Results demonstrated the formation of covalent adduction products between biological cysteine thiols and reactive moieties on cocaine and morphine metabolites. Rigorous mass spectrometric analysis in conjunction with in vitro metabolic activation, pharmacogenetic reaction phenotyping, and computational modeling were utilized to characterize structures and mechanisms of formation for each resultant thiol adduction product. For cocaine, data collected demonstrated the formation of adduction products from a reactive arene epoxide intermediate, designating a novel metabolic pathway for cocaine. In the case of morphine, data expanded on known adduct-forming pathways using sensitive and selective analysis techniques, following the known reactive metabolite, morphinone, and a proposed novel metabolite, morphine quinone methide. Data collected in this study describe novel metabolic events for multiple important drugs of abuse, culminating in detection methods and mechanistic descriptors useful to both medical and forensic investigators when examining the toxicology associated with cocaine, methamphetamine, and morphine.

Comprehensive forensic toxicological analysis of designer drugs

New designer drugs are constantly emerging onto the illicit drug market and it is often difficult to validate and maintaincomprehensive analytical methods for accurate detection of these compounds. Generally, toxicology laboratories utilize a screening method, such as immunoassay, for the presumptive identification of drugs of abuse. When a positive result occurs, confirmatory methods, such as gas chromatography (GC) or liquid chromatography (LC) coupled with mass spectrometry (MS), are required for more sensitive and specific analyses. In recent years, the need to study the activities of these compounds in screening assays as well as to develop confirmatory techniques to detect them in biological specimens has been recognized. Severe intoxications and fatalities have been encountered with emerging designer drugs, presenting analytical challenges for detection and identification of such novel compounds. The first major task of this research was to evaluate the performance of commercially available immunoassays to determine if designer drugs were cross-reactive. The second major task was to develop and validate a confirmatory method, using LC-MS, to identify and quantify these designer drugs in biological specimens.^ Cross-reactivity towards the cathinone derivatives was found to be minimal. Several other phenethylamines demonstrated cross-reactivity at low concentrations, but results were consistent with those published by the assay manufacturer or as reported in the literature. Current immunoassay-based screening methods may not be ideal for presumptively identifying most designer drugs, including the "bath salts." For this reason, an LC-MS based confirmatory method was developed for 32 compounds, including eight cathinone derivatives, with limits of quantification in the range of 1-10 ng/mL. The method was fully validated for selectivity, matrix effects, stability, recovery, precision, and accuracy. In order to compare the screening and confirmatory techniques, several human specimens were analyzed to demonstrate the importance of using a specific analytical method, such as LC-MS, to detect designer drugs in serum as immunoassays lack cross-reactivity with the novel compounds. Overall, minimal cross-reactivity was observed, highlighting the conclusion that these presumptive screens cannot detect many of the designer drugs and that a confirmatory technique, such as the LC-MS, is required for the comprehensive forensic toxicological analysis of designer drugs.^

Cocaine Enhances HIV-1 Infectivity in Monocyte Derived Dendritic Cells by Suppressing microRNA-155

Cocaine and other drugs of abuse increase HIV-induced immunopathogenesis; and neurobiological mechanisms of cocaine addiction implicate a key role for microRNAs (miRNAs), single-stranded non-coding RNAs that regulate gene expression and defend against viruses. In fact, HIV defends against miRNAs by actively suppressing the expression of polycistronic miRNA cluster miRNA-17/92, which encodes miRNAs including miR-20a. IFN-g production by natural killer cells is regulated by miR-155 and this miRNA is also critical to dendritic cell (DC) maturation. However, the impact of cocaine on miR-155 expression and subsequent HIV replication is unknown. We examined the impact of cocaine on two miRNAs, miR-20a and miR-155, which are integral to HIV replication, and immune activation. Using miRNA isolation and analysis, RNA interference, quantitative real time PCR, and reporter assays we explored the effects of cocaine on miR-155 and miR-20 in the context of HIV infection. Here we demonstrate using monocyte-derived dendritic cells (MDCCs) that cocaine significantly inhibited miR-155 and miR-20a expression in a dose dependent manner. Cocaine and HIV synergized to lower miR-155 and miR-20a in MDDCs by 90%. Cocaine treatment elevated LTR-mediated transcription and PU.1 levels in MDCCs. But in context of HIV infection, PU.1 was reduced in MDDCs regardless of cocaine presence. Cocaine increased DC-SIGN and and decreased CD83 expression in MDDC, respectively. Overall, we show that cocaine inhibited miR-155 and prevented maturation of MDDCs; potentially, resulting in increased susceptibility to HIV-1. Our findings could lead to the development of novel miRNA-based therapeutic strategies targeting HIV infected cocaine abusers.

The Effects of American Sign Language on General Self- Efficacy and Anxiety Among Mothers in a Residential Rehabilitation Facility for Drug Addiction and Substance Abuse

Globally, approximately 208 million people aged 15 and older used illicit drugs at least once in the last 12 months; 2 billion consumed alcohol and tobacco consumption affected 25% (World Drug Report, 2008). In the United States, 20.1 million (8.0%) people aged 12 and older were illicit drug users, 129 million (51.6%) abused alcohol and 70.9 million (28.4%) used tobacco (SAMHSA/OAS, 2008).Usually considered a problem specific to men (Lynch, 2002), 5.2% of pregnant women aged 15 to 44 are also illicit drug and substance abusers (SAMHSA/OAS, 2007). During pregnancy, illicit drugs and substance abuse (ID/SA) can significantly affect a woman and her infant contributing to developmental and communication delays for the infant and influencing parenting abilities (Budden, 1996; March of Dimes, 2006b; Rossetti, 2000). Feelings of guilt and shame and stressful experiences influence approaches to parenting (Ashley, Marsden, & Brady, 2003; Brazelton, & Greenspan, 2000; Ehrmin, 2000; Johnson, & Rosen, 1990; Kelley, 1998; Rossetti, 2000; Velez et al., 2004; Zickler, 1999). Parenthood is an expanded role that can be a trying time for those lacking a sense of self-efficacy and creates a high vulnerability to stress (Bandura, 1994). Residential treatment programs for ID/SA mothers and their children provide an excellent opportunity for effective interventions (Finkelstein, 1994; Social Care Institute for Excellence, 2005). This experimental study evaluated whether teaching American Sign Language (ASL) to mothers living with their infants/children at an ID/SA residential treatment program increased the mothers’ self-efficacy and decreased their anxiety. Quantitative data were collected using the General Self-Efficacy Scale and the State-Trait Anxiety Inventory showing there was both a significant increase in self efficacy and decrease in anxiety for the mothers. This research adds to the knowledge base concerning ID/SA mothers’ caring for their infants/children. By providing a simple low cost program, easily incorporated into existing rehabilitation curricula, the study helps educators and healthcare providers better understand the needs of the ID/SA mothers. This study supports Bandura’s theory that parents who are secure in their efficacy can navigate through the various phases of their child’s development and are less vulnerable to stress (Bandura, 1994).

Risk factors that predict maladaptive outcomes among sexually abused girls presenting for services at a community agency

The purpose of this study was to determine which factors predicted maladaptive outcomes in sexually abused children. Key factors were aggregated into four categories: abuse characteristics risk factors, individual-level risk factors, family disruption risk factors, and social disruption risk factors. It was hypothesized that (a) individual-level risk factors (e.g., school performance, child alcohol/substance abuse) and (b) abuse characteristics risk factors (e.g., longer duration/frequency of abuse, use of force/threats of force, intrafamilial abuse) would predict higher levels of trauma symptoms. Furthermore, it was hypothesized that (a) family disruption risk factors (e.g., family alcohol/substance use, family psychopathology) and (b) social disruption risk factors (e.g., parental divorce, homelessness, witnessing homicide or violence) would moderate the impact of prior sexual abuse and predict higher levels of trauma symptoms. ^ The participants were 110 female children (5 to 18 years old) presenting for treatment for sexual abuse at a community agency (The Journey Institute) in Miami, Florida. This study conducted a retrospective analysis of an archival data set collected over a three-year period (1998–2001). The measures completed upon intake included The Journey Psychosocial Assessment and The Trauma Symptom Checklist for Children (TSCC; Briere, 1996). Using Pearson correlations and hierarchical multiple regression analysis, this study found that abuse characteristics risk factors and individual-level risk factors were predictive of maladaptive outcomes in this sample of sexually abused girls. However, no moderating effects were found for family disruption risk factors or social disruption risk factors. Therefore, the results of these analyses provided support for the contention that abuse characteristics and individual-level risk factors were appropriate targets for treatment for sexually abused girls. Moreover, limitations of this study, implications for treatment, and directions for future research were discussed. ^

The effects of peer teaching of infant massage on general elf-efficacy and mother infant attachment among mothers in a residential rehabilitation facility for drug addiction and substance abuse

Approximately 200 million people, 5% aged 15-64 worldwide are illicit drug or substance abusers (World Drug Report, 2006). Between 2002 and 2005, an average of 8.2% of 12 year olds and older in the Miami, Fort Lauderdale metropolitan areas used illicit drugs (SAMHSA, 2007). Eight percent of pregnant women, aged 15 to 25, were more likely to have used illicit drugs during pregnancy than pregnant women aged 26 to 44. Alcohol use was 9.8% and cigarette use was 18% for pregnant women aged 15 to 44 (SAMHSA, 2005). Approximately a quarter of annual birth defects are attributed to the exposure of drugs or substance abuse in utero (General Accounting Office, 1991). Physical, psychological and emotional challenges may be present for the illicit drug/substance abuse (ID/SA) mother and infant placing them at a disadvantage early in their relationship (Shonkoff & Marshall, 1990). Mothers with low self efficacy have insecurely attached infants (Donovan, Leavitt, & Walsh, 1987). As the ID/SA mother struggles with wanting to be a good parent, education is needed to help her care for her infant. In this experimental study residential rehabilitating ID/SA mothers peer taught infant massage. Massage builds bonding/attachment between mother and infant (Reese & Storm, 2008) and peer teaching is effective because participants have faced similar challenges and speak the same language (Boud, Cohen, & Sampson 2001). Quantitative data were collected using the General Self-Efficacy and Maternal Attachment Inventory-Revised Scale before and after the 4-week intervention program. A reported result of this study was that empowering ID/SA mothers increased their self-efficacy, which in turn allowed the mothers to tackle challenges encountered and created feelings of being a fit mother to their infants. This research contributes to the existing database promoting evidence-based practice in drug rehabilitation centers. Healthcare personnel, such as nurse educators and maternal-child health practitioners, can develop programs in drug rehabilitation centers that cultivate an environment where the ID/SA rehabilitating mothers can peer teach each other, while creating a support system. Using infant massage as a therapeutic tool can develop a healthy infant and nurture a more positive relationship between mother and infant.

The effects of American Sign Language on general self-efficacy and anxiety among mothers in a residential rehabilitation facility for drug addiction and substance abuse

Globally, approximately 208 million people aged 15 and older used illicit drugs at least once in the last 12 months; 2 billion consumed alcohol and tobacco consumption affected 25% (World Drug Report, 2008). In the United States, 20.1 million (8.0%) people aged 12 and older were illicit drug users, 129 million (51.6%) abused alcohol and 70.9 million (28.4%) used tobacco (SAMHSA/OAS, 2008).Usually considered a problem specific to men (Lynch, 2002), 5.2% of pregnant women aged 15 to 44 are also illicit drug and substance abusers (SAMHSA/OAS, 2007). During pregnancy, illicit drugs and substance abuse (ID/SA) can significantly affect a woman and her infant contributing to developmental and communication delays for the infant and influencing parenting abilities (Budden, 1996; March of Dimes, 2006b; Rossetti, 2000). Feelings of guilt and shame and stressful experiences influence approaches to parenting (Ashley, Marsden, & Brady, 2003; Brazelton, & Greenspan, 2000; Ehrmin, 2000; Johnson, & Rosen, 1990; Kelley, 1998; Rossetti, 2000; Velez et al., 2004; Zickler, 1999). Parenthood is an expanded role that can be a trying time for those lacking a sense of self-efficacy and creates a high vulnerability to stress (Bandura, 1994). Residential treatment programs for ID/SA mothers and their children provide an excellent opportunity for effective interventions (Finkelstein, 1994; Social Care Institute for Excellence, 2005). This experimental study evaluated whether teaching American Sign Language (ASL) to mothers living with their infants/children at an ID/SA residential treatment program increased the mothers’ self-efficacy and decreased their anxiety. Quantitative data were collected using the General Self-Efficacy Scale and the State-Trait Anxiety Inventory showing there was both a significant increase in self efficacy and decrease in anxiety for the mothers. This research adds to the knowledge base concerning ID/SA mothers’ caring for their infants/children. By providing a simple low cost program, easily incorporated into existing rehabilitation curricula, the study helps educators and healthcare providers better understand the needs of the ID/SA mothers. This study supports Bandura’s theory that parents who are secure in their efficacy can navigate through the various phases of their child’s development and are less vulnerable to stress (Bandura, 1994).

``This is your brain on drugs'': The public policy of anti -drug programming. An exploratory analysis of media and other sources of drug information for adolescents

In the 1980s, government agencies sought to utilize research on drug use prevention to design media campaigns. Enlisting the assistance of the national media, several campaigns were designed and initiated to bring anti-drug use messages to adolescents in the form of public service advertising. This research explores the sources of information selected by adolescents in grades 7 through 12 and how the selection of media and other sources of information relate to drug use behavior and attitudes and perceptions related to risk/harm and disapproval of friends' drug-using activities.^ Data collected from 1989 to 1992 in the Miami Coalition School Survey provided a random selection of secondary school studies. The responses of these students were analyzed using multivariate statistical techniques.^ Although many of the students selected media as the source for most of their information on the effects of drugs on the people who use them, the selection of media was found to be positively related to alcohol use and negatively related to marijuana use. The selection of friends, brothers, or sisters was a statistically significant source for adolescents who smoke cigarettes, use alcohol or marijuana.^ The results indicate that the anti-drug use messages received by students may be canceled out by media messages perceived to advocate substance use and that a more persuasive source of information for adolescents may be friends and siblings. As federal reports suggest that the economic costs of drug abuse will reach an estimated $150 billion by 1997 if current trends continue, prevention policy that addresses the glamorization of substance use remains a national priority. Additionally, programs that advocate prevention within the peer cluster must be supported, as peers are an influential source for both inspiring and possibly preventing drug use behavior. ^

The Effects of Peer Teaching of Infant Massage on General Self-Efficacy and Mother Infant Attachment Among Mothers in a Residential Rehabilitation Facility for Drug Addiction and Substance Abuse

Approximately 200 million people, 5% aged 15-64 worldwide are illicit drug or substance abusers (World Drug Report, 2006). Between 2002 and 2005, an average of 8.2% of 12 year olds and older in the Miami, Fort Lauderdale metropolitan areas used illicit drugs (SAMHSA, 2007). Eight percent of pregnant women, aged 15 to 25, were more likely to have used illicit drugs during pregnancy than pregnant women aged 26 to 44. Alcohol use was 9.8% and cigarette use was 18% for pregnant women aged 15 to 44 (SAMHSA, 2005). Approximately a quarter of annual birth defects are attributed to the exposure of drugs or substance abuse in utero (General Accounting Office, 1991). Physical, psychological and emotional challenges may be present for the illicit drug/substance abuse (ID/SA) mother and infant placing them at a disadvantage early in their relationship (Shonkoff & Marshall, 1990). Mothers with low self efficacy have insecurely attached infants (Donovan, Leavitt, & Walsh, 1987). As the ID/SA mother struggles with wanting to be a good parent, education is needed to help her care for her infant. In this experimental study residential rehabilitating ID/SA mothers peer taught infant massage. Massage builds bonding/attachment between mother and infant (Reese & Storm, 2008) and peer teaching is effective because participants have faced similar challenges and speak the same language (Boud, Cohen, & Sampson 2001). Quantitative data were collected using the General Self-Efficacy and Maternal Attachment Inventory-Revised Scale before and after the 4-week intervention program. A reported result of this study was that empowering ID/SA mothers increased their self-efficacy, which in turn allowed the mothers to tackle challenges encountered and created feelings of being a fit mother to their infants. This research contributes to the existing database promoting evidence-based practice in drug rehabilitation centers. Healthcare personnel, such as nurse educators and maternal-child health practitioners, can develop programs in drug rehabilitation centers that cultivate an environment where the ID/SA rehabilitating mothers can peer teach each other, while creating a support system. Using infant massage as a therapeutic tool can develop a healthy infant and nurture a more positive relationship between mother and infant.