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.

Treatment attrition among racial /ethnic minority adolescents participating in a brief motivational intervention for substance use problems: The influence of individual, social, cultural, and treatment factors

The present study has the primary aim of examining the predictors of treatment attrition among racial/ethnic minority adolescents with substance use problems. This study explores the potential differential influence of specific individual, social, cultural, and treatment factors on treatment attrition within three racial/ethnic subgroups of adolescents. Participants: A unique feature of the study is the use of a racial/ethnic minority sample (N=453), [U.S.-born Hispanics (n = 262), Foreign-born Hispanics (n = 117), and African-Americans (n = 74)]. Multivariate logit analyses were used to examine the influence of specific factors on treatment attrition among the full sample of adolescents, as well as within each racial/ethnic subgroup. Consistent with expectations, multivariate logit analyses reveal that, the specific factors associated with attrition varied across the three racial/ethnic subgroups. Having parents with problem substance use, being on the waitlist, and being court mandated to treatment emerged as predictors of attrition among the US-born Hispanics, while only Conduct Disorder was significantly associated with greater attrition among foreign-born Hispanics. Finally, among African-Americans, parental crack/cocaine use, parental abstinence from alcohol, and being on the waitlist were predictive of attrition. Multiple factors were associated with treatment attrition among racial/ethnic minority adolescents with specific factors differentially predicting attrition within each racial/ethnic subgroup. African-American youth were more than twice as likely as their Hispanic counterparts to leave treatment prematurely. It is critically important to understand the predictors of attrition among racial/ethnic minority youth in order to better meet the needs of youth most at risk of dropping out. ^

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.

Does U.S counter-drug policy affect nationalism in the Anglophone Caribbean? A comparative study on the impact of counter-drug policy on nationalism in Jamaica and Trinidad and Tobago

This dissertation examined the effect of United States counter-drug policy on nationalism in small states, focusing on Jamaica and Trinidad and Tobago. The states were selected for their roles and geostrategic importance in the illegal drug trade; Jamaica being the largest drug producing country in the Anglophone Caribbean and having strong links to the trade of Colombian cocaine, and Trinidad being a mere seven miles from the South American coast. Since U.S. counterdrug policies have frequently been viewed in the region as imperialistic, this dovetails into ideas on the perceptions of smallness and powerlessness of Caribbean nations. Hence, U.S. drug policies affect every vulnerability faced by the Caribbean, individually and collectively. Thus, U.S. drug policy was deemed the most appropriate independent variable, with nationalism as the dependent variable. In both countries four Focus Groups and one Delphi Study were conducted resulting in a total of 60 participants. Focus Group participants, recruited from the general population, were asked about their perception of the illegal drug trade in the country and the policies their government had created. They were also asked their perception on how deeply involved the U.S. was in the creation of these policies and their opinions on whether this involvement was positive or negative. The Delphi Study participants were experts in the field of local drug policies and also gave their interpretations of the role the U.S. played in local policy creation. Coupled with this data, content analysis was conducted on various newspaper articles, press releases, and speeches made regarding the topic. In comparing both countries, it was found that there is a disconnect between government actions and the knowledge and perceptions of the general public. In Trinidad and Tobago this disconnect was more apparent given the lack of awareness of local drug policies and the utter lack of faith in government solutions. The emerging conclusion was that the impact of U.S. drug policy on nationalism was more visible in Trinidad and Tobago where there was a weaker civil society-government relationship, while the impact on nationalism was more obscure in Jamaica, which had a stronger civil-society government relationship.

Hispanic Adolescents With Severe Substance Abuse Issues: Parental Involvement, Acculturation-Related Factors and Attachment

The main objective of the study was to investigate the relationship between parent-related, acculturation-related, and substance use-related variables found within individual, familial/parental, peer and school adolescent ecological domains, in a clinical sample (i.e. adolescents who met criteria for a Diagnostic Statistical Manual-IV [DSM-IV] clinical diagnosis of substance abuse/dependence) of Hispanic adolescents from Miami, Florida. The sample for this study consisted of 94 adolescent-mother pairs. The adolescent sample was 65% male, and 35% female, with a mean age of 15 years. More than half of the adolescents were born in the United States (60%) and had resided in the U.S. for an average of 12 years; 80% of the caregivers (primarily mothers) were foreign-born and lived in the U.S. for an average of 21 years. Correlation and hierarchical regression were used to answer the research questions. The findings indicate that the hypothesized model and corresponding anticipated effect of the relationship between parental school and peer involvement on adolescents’ frequency of alcohol, marijuana and cocaine use was not supported by the data. Parental “acculturation-related” variables did not explain any of the variance in adolescent substance use frequency in this sample. Mediation and moderation models were not supported either. However, some interesting relationships were found: The larger the acculturation gap, the lower the parental involvement in school tended to be (r = -.21, p < .05). Adolescents who experienced a greater acculturation gap with their parents (-.81, p >.01) had an earlier onset of marijuana (-.33, p < .01) and cocaine use (r = -.24, p .05), they also reported using marijuana more frequently than females (.21, p >.05).

Treatment attrition among racial/ethnic minority adolescents participating in a brief motivational intervention for substance use problems : the influence of individual, social, cultural, and treatment factors

The present study has the primary aim of examining the predictors of treatment attrition among racial/ethnic minority adolescents with substance use problems. This study explores the potential differential influence of specific individual, social, cultural, and treatment factors on treatment attrition within three racial/ethnic subgroups of adolescents. Participants: A unique feature of the study is the use of a racial/ethnic minority sample (N=453), [U.S.-born Hispanics (n = 262), Foreign-born Hispanics (n = 117), and African- Americans (n = 74)]. Multivariate logit analyses were used to examine the influence of specific factors on treatment attrition among the full sample of adolescents, as well as within each racial/ethnic subgroup. Consistent with expectations, multivariate logit analyses reveal that, the specific factors associated with attrition varied across the three racial/ethnic subgroups. Having parents with problem substance use, being on the waitlist, and being court mandated to treatment emerged as predictors of attrition among the US-born Hispanics, while only Conduct Disorder was significantly associated with greater attrition among foreign-born Hispanics. Finally, among African-Americans, parental crack/cocaine use, parental abstinence from alcohol, and being on the waitlist were predictive of attrition. Multiple factors were associated with treatment attrition among racial/ethnic minority adolescents with specific factors differentially predicting attrition within each racial/ethnic subgroup. African-American youth were more than twice as likely as their Hispanic counterparts to leave treatment prematurely. It is critically important to understand the predictors of attrition among racial/ethnic minority youth in order to better meet the needs of youth most at risk of dropping out.

The Construction and Optimization on an Ion Mobility Spectrometer for the Analysis of Explosives and Drugs

Today, over 15,000 Ion Mobility Spectrometry (IMS) analyzers are employed at worldwide security checkpoints to detect explosives and illicit drugs. Current portal IMS instruments and other electronic nose technologies detect explosives and drugs by analyzing samples containing the headspace air and loose particles residing on a surface. Canines can outperform these systems at sampling and detecting the low vapor pressure explosives and drugs, such as RDX, PETN, cocaine, and MDMA, because these biological detectors target the volatile signature compounds available in the headspace rather than the non-volatile parent compounds of explosives and drugs. In this dissertation research volatile signature compounds available in the headspace over explosive and drug samples were detected using SPME as a headspace sampling tool coupled to an IMS analyzer. A Genetic Algorithm (GA) technique was developed to optimize the operating conditions of a commercial IMS (GE Itemizer 2), leading to the successful detection of plastic explosives (Detasheet, Semtex H, and C-4) and illicit drugs (cocaine, MDMA, and marijuana). Short sampling times (between 10 sec to 5 min) were adequate to extract and preconcentrate sufficient analytes (> 20 ng) representing the volatile signatures in the headspace of a 15 mL glass vial or a quart-sized can containing ≤ 1 g of the bulk explosive or drug. Furthermore, a research grade IMS with flexibility for changing operating conditions and physical configurations was designed and fabricated to accommodate future research into different analytes or physical configurations. The design and construction of the FIU-IMS were facilitated by computer modeling and simulation of ion’s behavior within an IMS. The simulation method developed uses SIMION/SDS and was evaluated with experimental data collected using a commercial IMS (PCP Phemto Chem 110). The FIU-IMS instrument has comparable performance to the GE Itemizer 2 (average resolving power of 14, resolution of 3 between two drugs and two explosives, and LODs range from 0.7 to 9 ng). The results from this dissertation further advance the concept of targeting volatile components to presumptively detect the presence of concealed bulk explosives and drugs by SPME-IMS, and the new FIU-IMS provides a flexible platform for future IMS research projects.

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.

Caffeine Intake and its Association with Disease Progression, Sleep Quality and Anxiety Symptoms and Nutritional Alterations in People Living with HIV in the Miami Adult Studies on HIV Cohort

Miami-Dade County has approximately 27,000 people living with HIV (PLWH), and the highest HIV incidence in the nation. PLWH have reported several types of sleep disturbances. Caffeine is an anorexic and lipolytic stimulant that may adversely affect sleep patterns, dietary intakes and body composition. High caffeine consumption (>250 mg. per day or the equivalent of >4 cups of brewed coffee) may also affect general functionality, adherence to antiretroviral treatment (ART) and HIV care. This study assess the relationship of high caffeine intake with markers of disease progression, sleep quality, insomnia, anxiety, nutritional intakes and body composition. A convenience sample of 130 PLWH on stable ART were recruited from the Miami Adult Studies on HIV (MASH) cohort, and followed for three months. After consenting, questionnaires on Modified Caffeine Consumption (MCCQ), Pittsburg Insomnia Rating Scale (PIRS), Pittsburg Sleep Quality Index (PSQI), Generalized Anxiety Disorder-7 (GAD-7), socio-demographics, drug and medication use were completed. CD4 count, HIV viral load, anthropometries, and body composition measures were obtained. Mean age was 47.89±6.37 years, 60.8% were male and 75.4% were African-Americans. Mean caffeine intake at baseline was 337.63 ± 304.97 mg/day (Range: 0-1498 mg/day) and did not change significantly at 3 months. In linear regression, high caffeine consumption was associated with higher CD4 cell count (β=1.532, P=0.049), lower HIV viral load (β=-1.067, P=0.048), higher global PIRS (β=1.776, P=0.046), global PSQI (β=2.587, P=0.038), and GAD-7 scores (β=1.674, P=0.027), and with lower fat mass (β=-0.994, P=0.042), energy intakes (β=-1.643, P=0.042) and fat consumption (β=-1.902, P=0.044), adjusting for relevant socioeconomic and disease progression variables. Over three months, these associations remained significant. The association of high caffeine with lower BMI weakened when excluding users of other anorexic and stimulant drugs such as cocaine and methamphetamine, suggesting that caffeine in combination, but not alone, may worsen their action. In summary, high caffeine consumption was associated with better measures of disease progression; but was also detrimental on sleep quality, nutritional intakes, BMI and body composition and associated with insomnia and anxiety. Large scale studies for longer time are needed to elucidate the contribution of caffeine to the well-being of PLWH.