Qualitative identification of fentanyl and other synthetic opioids using ambient ionization high resolution time-of-flight mass spectrometry

The Centers for Disease Control and Prevention deemed the increase in overdose fatalities, due to the use of opioids, an “opioid epidemic” in the United States. Heroin, fentanyl, and other synthetic opioids are commonly abused and are contributing to the opioid epidemic. In 2016, the Drug Enforcement Administration temporarily placed three fentanyl analogs (beta-hydroxythiofentanyl, butyryl fentanyl, and furanyl fentanyl) under Schedule I due to their imminent threat to public health. These drugs elicit analgesic effects similar to heroin making them desirable drugs to abuse. Novel fentanyl analogs and designer opioids are expected to become more prominent in forensic casework in the near future as the opioid epidemic continues. These drugs can be seen in forensic seized drug and urine casework samples either alone or mixed with other drugs of abuse. It is therefore necessary to have an efficient methodology to identify these new compounds. Currently, gas chromatography-mass spectrometry (GC/MS) is used to identify drugs of abuse and is considered the “gold standard” in forensic casework. However, analysis times can often range from 15 to 60 minutes in length. Another drawback is the need for spectral library matching, which requires analytical reference materials for identification. Therefore, the identification of novel fentanyl analogs and designer drugs is limited until a reference material becomes available. In this study, direct sample analysis time-of-flight mass spectrometry (DSA-TOFMS) was evaluated to provide rapid identification of fentanyl and other synthetic opioids in seized drug and urine casework samples. DSA is a direct ambient ionization source, which requires no chromatography and minimal sample preparation. TOFMS is a high resolution mass spectrometer that uses collision-induced dissociation (CID) to produce precursor ion and characteristic fragmentation ions, which provide additional structural and molecular formula information, allowing for the identification of compounds without a reference material. The analytes explored in this study include: heroin, 6-monoacetylmorphine (6-MAM), morphine, fentanyl, norfentanyl, 4-anilino-N-phenethylpiperidine (4-ANPP), acetyl fentanyl, beta-hydroxythiofentanyl, butyryl fentanyl, furanyl fentanyl, valeryl fentanyl, AH-7921, U-47700, buprenorphine, norbuprenorphine, desomorphine, MT-45, W-15, and W-18. Direct sample analysis time-of flight mass spectrometry (DSA-TOFMS) is a novel instrumentation that could be utilized in the forensic sciences field to qualitatively identify illicit substances in casework samples. In this study, 19 compounds of interest containing heroin, fentanyl, fentanyl analogs, and other synthetic opioids were evaluated using DSA-TOFMS. DSA-TOFMS abbreviated the workload of the analysis and was utilized to provide precursor ion and characteristic fragmentation ions within an analysis time of 20 seconds. Certified reference standards were used to optimize instrumentation settings, to determine precursor ions and characteristic fragmentation ions, and to determine the limit of detection of the instrument. A carryover study determined there were no persisting ions present when entering the capillary inlet between runs. A repeatability study revealed the DSA-TOFMS repeated results within the acceptable criteria range of above 500 counts and within 10ppm error 93% (10ppm) and 83% (1ppm). Forensic seized drug casework samples were evaluated with DSA-TOFMS and qualitatively identified. Out of the 64 samples, 89% were qualitatively identified as heroin, 4% were qualitatively identified as fentanyl, 1% was qualitatively identified as heroin and fentanyl, 3% were qualitatively identified as acetyl fentanyl, and 3% were qualitatively identified as furanyl fentanyl. The casework samples containing furanyl fentanyl were considered “true unknown unknown samples,” as the Maine Health and Environmental Testing Laboratory gas chromatography-mass spectrometry library did not have a spectrum to use for the identification of these samples. Forensic urine casework samples were evaluated with DSA-TOFMS. Samples previously confirmed to contain compounds of interest were prepared using minimal sample preparation technique (filtered using 0.45 microns syringe filters and diluted (1:10) with LC/MS grade water). Analysis displayed the limitations of DSA-TOFMS as only twelve of the forty compounds of interest were present and only three of the twelve were within the acceptable criteria range. DSA-TOFMS is a fast and reliable technique with minimal sample preparation for forensic seized drug samples. However, the concentration in complex matrixes, such as urine and blood, were unable to be qualitatively identified using this sample preparation method by DSA-TOFMS

The Organization of American States (OAS) in Rhetoric and Reality

The purpose of this study is to examine how well the countries in the Western Hemisphere translate Organization of American States’ (OAS) resolutions into actual meaningful legislation, and how international discourse influences, or does not, domestic policy. This study will utilize the data program QDA Miner in order to better analyze texts of agreements and treaties put forth by the OAS, and to highlight the correlation between different types of rhetoric and meaningful member state action. Data will be gathered primarily from the OAS’ own data bases and compiled into the QDA software for analysis. This analysis will allow the agreements to be divided into “rhetoric categories.” After the agreements are analyzed and divided up into their respective rhetoric categories, regression will be run using SPSS 19.1 IBM software. This software will allow for the interpretation of whether or not there is a correlation between competing types of international discourse and the domestic policies of member states. Prior to this study, many different research studies have been carried out to examine what compels different countries act in accordance with International Organizations and what does not. Compliance with international declarations and agreements has long since been a question and focal point of study when analyzing international institutions and this study aims to understand what role, if any, language and rhetoric plays in the issue of compliance

Analysis of beta-band MEG coherence in ASD during direct gaze processing: Relationship to social cognition

Gaze-following is a rudimentary behavior that forms the foundation of social communication, where aberrant social orienting is a defining feature of Autism Spectrum Disorders (ASD; Hoehl et al., 2009; Nummenmaa & Cal der, 2009). Recent neuroimaging research has demonstrated increasing precision at identifying aberrant brain response patterns in individuals with ASD, but no studies have employed a more holistic neural network approach analyzing coherence (i.e., synchrony of neural oscillations) during direct gaze processing. The current study examined coherence between each pair of 54 brain regions and the relationship between average coherence and psychometric measures of social cognition in eleven participants with ASD and eight typically developing (TD) controls, who passively viewed direct gaze while undergoing Magnetoencephalography (MEG). Results revealed significant intra- and inter-hemispheric between-group differences in average coherence (1-45 Hz), providing preliminary support for increased long-range left hemisphere coherence and increased interhemisphere occipital-occipital activity in individuals with ASD

Identification of Antigens of Edwardsiella Ictaluri, the Causative Agent of Enteric Septicemia of Catfish (ESC).

Edwardsiella ictaluri, causative agent of enteric septicemia of catfish (ESC), is the primary bacterial pathogen of commercially produced channel catfish (Ictalurus punctatus). The purpose of this study was to make progress towards development of a successful ESC vaccine by generating and characterizing a pool of E. ictaluri antigens and determining if they were protective against ESC in catfish. Antigenic protein expression was evaluated in E. ictaluri cells under different conditions of growth using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and western blotting with pooled convalescent catfish serum (CCS). Results showed protein bands separated by one-dimensional PAGE may consist of one or more antigenic and non-antigenic proteins, and strain and culture temperatures do not effect expression of antigenic proteins, but culture media does. A 57 KD antigenic protein was only expressed by E. ictaluri grown in minimal media, and antigenic proteins of 58 and 71 KD were expressed at different levels in minimal or rich media. An E. ictaluri genomic library in an Escherichia coli expression vector was screened using goat anti-E. ictaluri serum (GAI) resulting in isolation of 32 clones expressing antigenic E. ictaluri proteins. Encoded genes and expressed antigenic proteins of nine clones were analyzed by partial DNA sequencing, 2D-PAGE and western blotting with GAI and CCS. The DNA inserts of three clones were double-strand sequenced. Four putative open reading frames were identified in the insert of clone 4d6 corresponding to antigenic proteins of 63, 20 and 18 KD expressed by both the clone and E. ictaluri cells. Genes encoding these proteins had no homology with other known genes. Overlapping inserts of clones 5d2 and 5d3 encoded homologs of E. coli partial genes serA and pgk, and complete genes rpiA, iciA, yggE, yggB and fda. Cloned antigenic E. ictaluri proteins of 33, 27, 35 and 45 KD were putatively identified as products of yggE, rpiA, iciA and fda respectively. Protective capabilities of vaccines of cloned antigenic proteins were evaluated in catfish. All vaccine treatments were protective against E. ictaluri challenge, however results were inconclusive due high levels of cross-reactive protection of E. coli, host strain of the cloning vector

A Comparison of Methods to Fit a Model to Simultaneous Time Series

This research project determines which methods are the most effective for finding a best fit model for simultaneous time series. The type of model used was an Autoregressive Integrated Moving Average (ARIMA) model. Two distinct methods were used when determining what order to assign to the ARIMA model: 1.) using the floor of the average number of autoregressive and moving average terms, and 2.) using the ceiling of the average number of autoregressive and moving average terms. After fitting the model, the Akaike Information Criterion (AIC) value for each method measured the goodness of fit to compare to fitting separate models to each series. Based on the results of this research the most effective method depends on the type of data that is being fitted. In most of the different cases explored, the floor function method and the ceiling function method had very similar results. However, for two specific cases the ceiling function was the more effective method. Therefore, it is important to consider the characteristics of the data that is being fitted to determine the most effective method

Influences of censorship challenges on state textbook adoption criteria which affect school curriculum

This study explored censorship in the public schools through the analysis of the changing textbook adoption criteria of the twenty-two adoption states to determine if new, subtle challenges of censorship suggest previously undiscovered effects on the established school curriculum. Adoption criteria (1975-76 and 1985-86) from the twenty-two adoption states were requested. State statutes were also reviewed. A content analysis focusing on the specified criteria needed for text adoption and the amount of public participation in the adoption process was conducted. It was found that few requirements have been added to the textbook adoption criteria since 1975 although some changes have occurred. Nineteen states have some type of specified criteria either in their state statues or state board of education policies. Sixteen states include some aspect of public participation in their textbook adoption process. Conclusions are: (1) The term state adoption state is ambiguous and the states comprising that group are not well-defined; (2) There is little consistency in the expectations for texts and the degree of public involvement in these states\u27 adoption process; (3) Additional specified criteria appear to be being added as state board of education policy rather than state statutes; and (4) More public participation is occurring in 1985 than did in 1975 and is happening in a variety of ways. Recommendations include: (1) State adoption state should be precisely defined in future study and a poll of all states taken to actually determine which states should be so labeled; (2) States with specified criteria for text adoption should review them for nebulous guidelines and provide standards for evaluation. States without criteria should review their adoption process to assess what is happening in practice; (3) Where oral testimony is accepted as part of the adoption process, written testimony should be included; (4) Textbook reviewers should receive training about state expectations before review process; (5) A pro-active stance on making the public aware of the textbook adoption process could occur if the adoption states form a public awareness coalition. Censorship awareness programs could be provided by the state for local districts; (6) Study of the textbook adoption process should continue with regards to censorship

Finding and Organizing Tolkien’s Invented Languages

Finding and Organizing Tolkien’s Invented Languages Paper by Eileen Marie Moore. Tech Mod: Leslie Donovan

Investigation of the Hydrogeology of the Memphis Light, Gas, and Water Shaw Wellfield, Shelby County, Tennessee

The dynamics of sources and pathways of recharged water to the semi-confined Memphis and confined Fort Pillow aquifers were investigated to assess the sustainability and vulnerability of groundwater resources within the Shaw wellfield located in Memphis, Tennessee. Geochemical and environmental tracer data were used to study compartmentalized groundwater flow in the Memphis and Fort Pillow aquifers, assess the vulnerability of the Memphis aquifer to modern water recharge (\u3c60 years old), and to determine the source(s) and pathways of modern water recharge. Hydrologic tracer data indicate wells in the northwest corner of Shaw wellfield are vulnerable to modern water recharge, with wells 704 and 708 having up to 17% component of modern water. Isotopic data show the Fort Pillow aquifer water has an older and chemically distinct source from the Memphis aquifer, which supports groundwater flow to the Fort Pillow aquifer is compartmentalized from that in the Memphis aquifer

Child Maltreatment and Physical Health in Emerging Adulthood: Relationships with Physical Activity and Health Risk Behaviors

Child maltreatment is highly associated with physical health problems throughout the lifespan (Schafer, Morton, & Ferraro, 2014; Springer, Sheridan, Kuo, & Carnes, 2007). Yet much is still unknown regarding the trajectory of health problems including the onset of health risk indicators in emerging adulthood. Emerging adulthood also represents a unique timeframe with little demand for healthy behaviors. There is evidence that child maltreatment increases the involvement in health-risk behaviors such as substance use, risky sex, and binge eating (Downey, Gudmunson, Pang, & Lee, 2017; Röhr et al., 2015). The primary purpose of the current study was to examine if emerging adults who self-report higher levels of child maltreatment (CM) exhibit poorer physical health outcomes. These health outcomes included heart rate, obesity level, blood pressure, blood sugar, waist circumference, symptoms of illness, and perceived health. The second aim of this research was to explore the role that Physical Activity (PA) and health risk behaviors have in the trajectory of child maltreatment and poor adult health. Emerging adults in college (N=100) completed a physical health assessment, selfreported measures of health, health risk behaviors, and child maltreatment, and wore an accelerometer for 10 days to track PA. Results of this study revealed that child maltreatment scores predicted higher resting heart rate and increased symptoms of illness. Additionally, females with maltreatment history presented higher levels of obesity and higher Metabolic Syndrome conditions than their peers. When examining moderators of CTQ and health outcomes, PA moderated both resting heart rate and symptoms of illness while binge eating behavior moderated symptoms of illness. Results support the importance of learning more regarding the trajectory of child maltreatment to chronic disease as health risk indicators are present in emerging adults. Furthermore, results provide examples of health behaviors that may alter the effects of child maltreatment on current health. These include sedentary behavior and binge eating behavior exacerbating poor health symptoms, low intensity PA improving health symptoms, and higher levels of overall PA protecting resting heart rate