Spotting the diffusion of New Psychoactive Substances over the Internet

Online availability and diffusion of New Psychoactive Substances (NPS) represent an emerging threat to healthcare systems. In this work, we analyse drugs forums, online shops, and Twitter. By mining the data from these sources, it is possible to understand the dynamics of drugs diffusion and their endorsement, as well as timely detecting new substances. We propose a set of visual analytics tools to support analysts in tackling NPS spreading and provide a better insight about drugs market and analysis

Monitoring and Analysis of Novel Psychoactive Substances in Trends Databases, Surface Web and the Deep Web, with Special Interest and Geo-Mapping of the Middle East

BACKGROUND Novel or new psychoactive substances (NPS), also known as designer drugs and research chemicals, represent a relatively recent phenomenon which can be traced back to the last decade or even earlier. The growth of this phenomenon and its electronic trade (e-trade) has been logarithmic and alarming; its aftermaths are not limited to; the economy, individual and public health, or illicit drug trade. The discipline of NPS has been extensively studied since 2010. However, there are still deficits in; data from the Middle East and the developing world including Arabic countries (1), application of data science and inferential hypothesis testing (2), implementation of the principles and theories of social science (3), utilization of experimental designs including randomised controlled trials (RCT) and quasiexperimental studies (4), and ultimately the enactment of real-time web analysis and the realization of tools of knowledge discovery in databases (5). AIM AND OBJECTIVES This study will implement an innovative research approach by combining observational analyses and data science; the aim is to provide generalizable (inferential) data in relation to NPS e-commerce activities on both divisions of the web, surface and deep. The pinnacle objective is to; assess the proportional magnitude of NPS e-commerce activity in the Middle East (1), provide a thorough analysis of the e-vendors on the darknet, both globally and regionally (Middle East) (2), correlate change in trends of e-commerce with time (3), provide recommendations for future studies in relation to the ecommerce activity in the Middle East (4), and to discuss the colossal potential of data mining technologies (5). MATERIALS AND METHODS This dissertation embodies the integrative and combinatorial approach towards the investigation of the e-trade (e-commerce) of NPS; it is made of integrated studies allocated into eleven results chapters. The utilised investigative tools represent a mixed-breed of observational web analytics including; literature review (1), cross-sectional studies and surveys (2, 3), internet snapshots (4), retrospective analyses (5), and critical appraisal (6). These analyses took place in both appendices of the web (surface web and the anonymous deep web); the analyses specifically involved; Google Trends database (1), literature databases (2), drug fora (3), social communication e-media (3), news and media networks (4), Grams search engine of the deep web (5), the darknet and its e-marketplace (6), Alphabay, Agora, Valhalla, Hansa, other dedicated e-markets for NPS e-trade (7). Additional extrapolations were concluded via the use of surveys and e-surveys in a population of medical students from Iraq. The potentials for knowledge discovery in databases (KDD) were also discussed in all chapters. Each chapter was thoroughly investigated via; data science tools (I), inferential statistics and hypothesis testing (II). The latter was dependent on using the Microsoft Excel 2016, the Statistical Package for the Social Sciences (SPSS), and some online tools of data science. RESULTS AND DISCUSSION A systematic review of approximately 600 PubMed-indexed articles of NPS literature showed; attempts of NPS research started to evolve after 2010, almost one-third of the research output (36%) was of relevance to toxicology and analytic chemistry, while reviews and cross-sectional studies were less common (15%, 18%). The analysis of the individual basis of power showed that NPS researchers, legislators, and policymakers are lagging behind, whereas terrorist possesses the highest possible power. Power scores of e-vendors scored highest in the UK, US, and eastern Europe, while being almost absent in the Middle East. The complimentary usage of PubMed, drug fora, and Google Trends was successful in extrapolating the most trending and high-risk NPS; the contribution from the Middle East to incidents of intoxications and fatalities was absent except for Israel. Deep web analysis, including the darknet emarketplace, has shown that the contribution of the Middle East never exceeded 7% of the total etrade, data were limited to; Iran, Israel, Turkey, Afghanistan, Oman, United Arab Emirates, and Saudi Arabia. Other Arabic countries included; Egypt, Morocco, and Algeria. It was interesting to observe the e-vendors of NPS operating in the Middle East were highly involved in e-trade activities in other nations, primarily; the UK, Western Europe and Scandinavia, US, Canada, Australia, and New Zealand. Surveys and internet snapshots unveiled the lack of awareness and very low prevalence of (ab)use of NPS within the selected Iraqi population. Captagon was highly prevalent in the Middle East, unlike NBOMe and octodrine. In summary, the contribution from the Middle East was microscopic when compared to the developed world; it did not exceed 7% of the entire NPS phenomenon e-trade. Similarly, the NPS research in the region of the Middle East can be described to be in its infancy. The overall level-of-evidence of this dissertation is assumed to be of level-2b according to the classification system imposed by the Oxford Center for Evidence-Based Medicine (2009). CONCLUSION The growth of the NPS phenomenon, including the e-commerce and its links to terrorism, are reaching unprecedented levels. Unless some reasonable efforts and ingenious upgrades of the current research methodologies, the NPS trade and e-trade will continue to prevail rendering all its counter-attempts fade into dust; these attempts are not only limited to NPS research but also into; legislative actions, policy planning, and counter-terrorism. Upgrades should affect these front lines; increasing the quality and quantity of studies in developed countries including Middle Eastern and Arabic countries (1), incorporation of efficient use of data science and advanced web analytics (2), compulsory training of data science, biostatistics, and basic neuroscience for all NPS researchers, chemists, and toxicologists (3), validation and incorporation of data mining and real-time analyses (4), inclusion of the rarely-used experimental studies including RCTs, pragmatic RCTs, and animal modelling (5), enhancement and potentiation of internet snapshot techniques (6), and full exploitation of trends databases of the surface web (7). Perhaps, the integration of real-time data mining and data crunching, and inferential data science technique will represent the climax armament to antagonise the alarming e-trade

Forensic electrochemistry: developing electrochemical sensors for the detection of illicit compounds

The following thesis reports the development of novel electrochemical protocols to further expand the niche research area of Forensic Electrochemistry. 0 introduces the key fundamental concepts within electrochemistry detailing why it is a significant analytical tool. Also described within this chapter are prior examples of electrochemistry used within in a forensic environment to further justify the use of such techniques in this work setting a solid foundation for the development of electrochemical sensors for previously un-detected (electrochemically) materials. Chapter 2 focuses on the growing epidemic of “Legal Highs” formally known as “Novel (New) Psychoactive Substances” (NPSs) that, at the time of this research is a major concern for drug authorities. Highlighted within is the multitude of existing techniques to analyse NPSs yet unable to simultaneous detect in-the-field with great sensitivity. Chapter 3 provides a summation of the materials employed in this research in addition to the experimental procedures. Furthermore, a brief synopsis of the screen-printing methodology is provided in order to deliver further understanding of the novel electrochemical sensors that are used throughout the thesis. Chapter 4 explores the use of screen-printed electrodes) as a novel electrochemical sensor for illicit compounds; with a particular focus on the NPS mephedrone (4-MMC; MKat; the most commonly abused NPS). The common ‘date-rape’ drug Rohypnol® (flunitrazepam) is also detected using screen-printed electrodes for the first time. The concept of screen-printed electrodes as a novel detector for illicit materials is expanded within Chapter 5 exploring different carbon materials utility as a sensor as well as the avant-garde field of study “Regal Electrochemistry” which utilises British Currency (GBP) to successfully quantify 4-MMC. Finally, Chapter 6 provides a summary and conclusion of the presented work highlighting the societal impact of such research whilst also posturing future work to ensure the field of Forensic Electrochemistry continues to grow

Mass Spectrometry Application Strategies of Dried Blood Spots Analysis

El análisis de manchas de sangre seca (DBS) es una tecnología de análisis bien conocida, cuyas primeras aplicaciones de rutina se remontan a la década de 1960. Los avances en los instrumentos de espectrometría de masas durante el siglo pasado, permitieron alcanzar la sensibilidad necesaria para abrir nuevos mercados. Sin embargo, algunas incertidumbres y la falta de comprensión de los métodos han impedido hasta el momento que la tecnología tenga una amplia aceptación en el mercado.En esta tesis doctoral se han desarrollado y validado diversas técnicas, condiciones y flujos de trabajo de análisis DBS, que demuestran la viabilidad y el potencial de aplicación de la tecnología DBS-LC-MS.Se han desarrollado métodos avanzados para su aplicación en el campo de la pediatría neonatal, en particular la prueba del talón en recién nacidos, donde se ha ampliado y estandarizado el panel de análisis.También se han investigado nuevos campos de aplicación, como la vigilancia de medicamentos terapéuticos y la toxicología forense. Se describe la aplicación de la farmacovigilancia remota de antirretrovirales en regiones de escasos recursos y se presentan nuevos enfoques analíticos para la vigilancia del abuso de alcohol. Finalmente, en esta tesis se ha introducido un método innovador con el que se pueden detectar más de 1.200 drogas ilícitas a partir de una sola gota de sangre.La investigación científica realizada se presenta en forma de compendio de publicaciones (6), que son incluidas en esta Memoria. Se adjuntan a modo de apéndice otros dos trabajos del candidato que no constan oficialmente en dicho compendio. Todos los trabajos constituyen una unidad temática coherente sobre la técnica de la DBS y su acoplamiento a la espectrometría de masas.This doctoral thesis is a compendium of dried blood spot (DBS) applications in the fields of newborn screening, forensic toxicology and therapeutic drug monitoring. DBS is a well-known analysis technology, which first routine applications date back to the 1960s. Advancements in mass spectrometry instruments during the last century, enabled to reach the required sensitivity to open up new markets. Some uncertainties and missing method understanding remain and this is holding back the technology from wide spread market acceptance. For the general scientific acceptance of this technology, several methods have been developed and validated within this work. Advanced methods for the field of newborn screening were developed, where the analysis panel has been extended and standardized. Goal of the first study was to transfer the amino acids and acyl carnitines analysis onto the automated DBS-MS 500 platform. Also, a steroid panel of 17OHP, cortisol and androstenedione was defined to exclude the 17OHP from the immune assay panel and to transfer this as well onto the fully automated DBS-LC-MS/MS. The conventional 17OHP enzyme-linked immunosorbent assay (ELISA) based on a manually punched DBS disc leads to a high percentage of false positive. First, cortisol increases when the babies are stressed, which also leads to elevated 17OHP concentrations and secondly, the ELISA has significant cross reaction potential with steroid sulfate which can be monitored with androstenedione. Both, the amino acid and acyl carnitine detection plus the integration of a steroid panel into the DBS-LC-MS/MS workflow was successfully achieved [1]. Newborn screening panels are not unified across borders and sometimes even differ within countries and laboratories. The implementation of the fully automated DBS platform could be a good starting point of standardization and unification of those programs. Here, a method based on an official newborn screening kit was introduced within this thesis. The analysis takes only 2 minutes per sample; however it is limited to amino acids and acyl carnitines only. The DBS extract is directly guided to the mass spectrometer without any column. This is a well-known procedure in newborn screening and allows speeding up the process to its optimum [2]. Therapeutic drug monitoring, especially in remote and rural areas is another upcoming application field of DBS. Several methods have been supported and co-authored, whereas the first method focuses of the three antiretroviral drugs nevirapine, efavirenz and lopinavir. The study highlights the transportation advantages of DBS, without any biohazard labeling neither requiring cooling chains. DBS were drawn in Tanzania, Africa, and sent to Switzerland where the samples went through different climate zones. Nonetheless, the samples showed very good results and stability of the monitored drugs [3]. We ran a follow up study on some of the samples approximately one year after the publication, where still all analytes could be recovered in the same concentration as published. Also the DBS sampling itself was investigated in a rural area of Tanzania [4] and the technique proved to be suitable. Another therapeutic drug, where a more efficient sampling process is required for mass drug administration (MDA) campaigns, is Ivermectin. This drug lowers the incidence of river blindness and lymphatic filariasis infections. Further, recent studies demonstrated that Ivermectin is also active against several other parasites and even against certain mosquitoes. As one of these mosquitoes is Anopheles gambiae, the major vector of malaria in Africa, Ivermectin could be mass administered to fight malaria. In other words, the drug makes the human blood lethal for the Anopheles gambiae and therefore reduces the number of vectors in an area. Still, more safety data is required from a larger population. Here, an according fully automated DBS method has been developed and validated. In addition, a comparison study to plasma samples, stability and hematocrit impact has been studied [5]. In forensic toxicology, it was shown that Dried Matrix Spot (DMS) is a suitable technology for large scale DBS applications. A fully automated method, where either 1200 drugs can be screened from a DBS and a quantitative follow up method focusing on the 28 most abundant drugs of abuse, was developed. This was documented within two publications, where the first publication was a proof of concept study to determine if the detection limits can be reached using the DBS-LC-MS/MS methodology [6]. The second, follow up publication was a specific method development and implementation. More than 1200 illicit drugs can be screened from a single DBS or Dried Urine Spots (DUS) within 20 minutes per sample using a Forensic Toxicology Database. The criteria of bio-analytical method validation guidelines were fulfilled, and the method was transferred into a routine laboratory successfully [7]. Furthermore, the discovery of phosphatidylethanol (PEth) as direct alcohol marker prolonged the window of detection for alcohol consumption to several weeks. PEth proved to be instable during storage of liquid blood samples. By using DBS sampling, this biomarker can be stabilized due to the inactivation of enzymatic activity. Also, for PEth, a fully automated DBS-LC-MS/MS was established for the determination of the two most abundant PEth homologs in a range from 20–1500 ng/mL. Automated DBS card handling and online solid phase extraction LC-MS/MS permits baseline separation and detection of PEth 16:0/18:1 and PEth 16:0/18:2 within 7 minutes per sample [8]. The methods from the various fields of application were presented at several conferences as oral presentations and posters, showing their interest from the scientific blood analysis community.<br /

The freedom of scientific research

Never have the scope and limits of scientific freedom been more important or more under attack. New science, from artificial intelligence to genomic manipulation, creates unique opportunities to make the world a better place. But it also presents unprecedented dangers, which many believe threaten the survival of humanity and the planet. This collection, by an international and multidisciplinary group of leading thinkers, addresses three vital questions: (1) How are scientific developments impacting on human life and on the structure of societies? (2) How is science regulated, and how should it be regulated? (3) Are there ethical boundaries to scientific developments in some sensitive areas (e.g. robotic intelligence, biosecurity)? The contributors are drawn from many disciplines, and approach the issues in diverse ways to secure the widest representation of the many interests engaged. They include some of the most distinguished academics working in this field, as well as young scholars

Assessing the Pharmacological Properties of Novel Psychoactive Substances (NPS) Identified Online: In Silico Studies on Designer Benzodiazepines and Novel Synthetic Opioids

Background By 2022, a total of 1,127 of Novel Psychoactive Substances (NPS) have been identified worldwide and officially reported by the United Nations Office on Drugs and Crime (UNODC) and the European Monitoring Centre for Drugs and Drug Addiction (EMCDDA). An analysis of the surface web via the use of a web crawler, NPSfinder®, indicated that the number of NPS could be almost four times higher than that known to both the UNODC and EMCDDA. This is of particular concern, especially if one considers the public health risks and harms associated with NPS use/abuse and the paucity of data related to their pharmacological/toxicity profiles. In particular, in the last few years two NPS classes, i.e. novel synthetic opioids (NSOs) and designer benzodiazepines (DBZDs) were associated with serious side-effects and life-threatening scenarios (i.e., fatalities and overdoses). Gaps in knowledge Hence, with online NPS numbers exceeding those reported by official sources, there is a strong need to address the gap in knowledge concerning the discrepancies between the online and the evidence based NPS market(s); as well as the gap in knowledge concerning lack of pharmacological profiles for most of the newly-identified NPS. Objectives This programme of research aimed to: use data available from NPSfinder®, the UNODC and EMCDDA to assess the current general NPS scenarios, and in particular for DBZDs and NSOs; use in silico computational techniques to predict the biological activity of the emerging NPS; use the predicted values to infer possible health threats associated with the consumption of these substances, underscoring which of the NPS identified online could indeed represent a serious threat to public health; assess the potential of in silico methodologies as preliminary risk assessment tools; and subsequently inform relevant stakeholders about the risks associated with these new NPS. Methods The NPSfinder® web crawler was used to identify NPS which are available/discussed online. A comparison with UNODC and EMCDDA databases was then carried out to assess the extent of the total NPS scenario, and the numbers of the NSOs and DBZDs classes. To appreciate and predict the biological activities of NSOs and DBZDs, in silico models (e.g., quantitative structure-activity relationship (QSAR), Molecular Docking (MD) and pharmacophore mapping) were used as reliable, time- and cost- effective alternatives to the classical approaches such as in vivo, in vitro or preclinical studies. Results and Discussion A total of 4,231 NPS were identified on the surface web, almost four times the numbers reported by both UNDOC and EMCDDA databases (circa 1,127). These results suggest how the online content analysis should be considered as an important source for the assessment of the NPS scenario. The same discrepancy in the total NPS numbers was observed for each NPS class and a total of 115 DBZDs and 371 NSOs were identified compared to 33 and 123 reported by the UNODC respectively. To assess pharmacological profiles of these NSOs and DBZDs identified online, specific QSAR models were developed in MOE® and Forge™. For the prediction of biological activities of DBZDs, the γ-aminobutyric acid A receptor (GABA-AR) was used; the mu opioid receptor (MOR) was used for the NSOs. In addition, for the DBZDs, a set of new potential ligands resulting from “scaffold hopping” exercises conducted with MOE® was also evaluated. The generated QSAR models returned good performance statistics confirming their strong reliability in predicting the biological activity of an unknown or a newly-identified molecule. The DBZDs predicted to be the most active were flubrotizolam, clonazolam, pynazolam and, fluclotizolam, consistently with reported literature and/or drug discussion forums. In particular with flubrotizolam and fluclotizolam, it was found they were discussed on drug fora but not previously identified either by the UNODC or EMCDDA (flubrotizolam only). This suggests the possible presence on the market of very potent NPS which are still unknown to international agencies, potentially representing a serious threat to public health. Worrisome results were also obtained for the class of NSOs, with the identification of new and potent analogues of carfentanyl (10,000 more potent than morphine), i.e., 2-methyl carfentanyl, n-methyl-carfentanyl and butyryl-carfentanyl. Moreover, the scaffold hopping exercise conducted for the DBZDs class, strongly suggested that structural replacement of the pendant phenyl moiety could increase biological activity and highlighted the existence of a still unexplored chemical space for this NPS class. The results obtained with QSAR analysis were supported by molecular docking exercises, which gave an indication of the binding affinity of these NPS towards their respective receptors. Moreover, the binding affinity of a set of DBZDs was assessed for the MOR, in an attempt to assess a possible multi-receptor activity of these molecules. Conclusions The online identification of a great number of NPS, including very potent central nervous system depressants, represents a serious challenge, in particular if one considers that DBZDs and NSOs are usually consumed either together or in combination with stimulants for recreational purposes and self-medication. The high numbers of available molecules, their patterns of use and the paucity of pharmacological data could lead to worrisome outcomes, including the synergy of each NPS class side-effects, which could (and are) increasing the likelihood of respiratory depression, coma, and deaths. To retrieve an extensive picture of the current NPS drug scenario, the online analysis has proven very useful, if not fundamental. Its ability to identify novel mentioned NPS, in a timely manner, makes it a very important tool for a range of activities, including informing law-enforcement and public health stakeholders, supporting the European and United Nations Early Warning Systems impacting and influencing law-making and guiding monitoring/surveillance. Moreover, in silico methodologies, proven as reliable tools for a fast prediction of biological activity, could be used in describing the activity/toxicity profile of novel NPS, aiming at supporting both law enforcement in scheduling process and public health stakeholders in drafting treatment/management educational packages. Finally, the combination of online and in silico analysis could support and improve the risk assessment procedures currently in place for NPS

An optimized and validated method for screening and quantification of commonly encountered stimulant drugs and selected metabolites in dried blood spot (DBS) samples by ultra performance liquid chromatography-quadrupole time of flight-high resolution mass spectroscopy (UPLC QTOF-HRMS) analysis

Blood microsampling techniques, like Dried Blood Spots (DBS), have seen recent development as an alternative to wet blood samples and offer numerous advantages. Quantification of ng/mLrange concentrations using DBS microvolumes (10-20 µL) is made possible with cutting edge instrumentation. Worldwide establishment of per se limits for drug impaired driving requires timely methods of sampling blood for accurate drug concentration measurements and interpretation. The method validation for the analysis of stimulants and metabolites in DBS samples by Ultra Performance Liquid Chromatography-Quadrupole Time of Flight-High Resolution Mass Spectroscopy (UPLC-QTOF-HRMS) is presented. Limits of detection and quantitation to 10 ng/mL was achieved. Method validation criteria was satisfied for 10 of 14 analytes. DBS drug stability over 8 weeks varied by analyte. DBS samples may assist in overcoming challenges in blood sampling as they are less invasive, easily transported and stored, and accurate drug quantitation even at low concentrations in DBS samples is possible.Master of Science (MSc) in Chemical Science

Characterization, Imaging and Quantitation of Small Molecules by Ambient Ionization Mass Spectrometry

For more than a century mass spectrometry has been a well-known technique in the field of chemical analytics. Its selectivity and sensitivity has made it popular in various fields. From analysis of pure organics, its use is still being explored in the analysis of biomolecules, either purified or direct from tissue sections. For analyzing these vast arrays of molecules, typically the front end is modulated depending upon the need of the user. For direct analysis of a sample of interest, ionization techniques such as DESI, PS, MALDI, etc are incorporated into the front of the mass spectrometer. In this work, the ambient ionization techniques, DESI-MS and PS-MS was modulated for characterization, imaging and quantification of small molecules. The use of DESI-MS and PS-MS in the field of forensics, microbiology and pharmaceutics is described. With the optimization of the front end modes, better sensitivity, selectively and robustness was ensured

An investigation of multiple natural origins of religion

This study attempts to trace how religion could have originated in prehistory and antiquity, out of natural human and prehuman behaviour, without requiring the reality of the supernatural.Religion is here defined as beliefs, conceptions, practices and roles concerned with the putative supernatural. A variety of manifestations or elements of religious belief and practice can be identified. It is proposed that they have separate origins. Examples of religious elements are: life after death, ghosts, sacrifice, priests, shamans, gods, demons, .... It is argued that to try to reduce religion to one original element is a mistake. There may be no single origin. But the individual elements have origins, and plausible theories can account for each.Using theories and insights of previous workers, elaborated as necessary with information from a range of sciences, arguments are presented to account for five major foundational religious elements, thereby illustrating and partly fulfilling what is potentially a much wider programme. The elements covered are: (1) Animatism: numina, daemons; (2) Animism: ghosts, souls; (3) Another world: life after death; (4) Another world: heaven; (5) Religious specialists: shamans.Chapter 1 introduces the programme. Chapter 2 sets out definitions, philosophical principles and methodologyChapter 3 explores the specifically numinous quality which characterizes the supernatural in subjective experience. Chapter 4 describes brain structures and the neural substrate of experience. Chapter 5 proposes specific neurological hypotheses to account for certain types of numinous or `supernatural' experience.Chapter 6 deals with ape mentality, which may be presumed to characterize that of our remote ancestors, and identifies precursors of religious elements.Chapters 7 - 11 deal with the possibly separate origin of five major religious elements, as listed above.Chapter 12 summarizes the investigation, attempts to place the elements covered in sequence of their development in prehistory and antiquity, and expresses the limitations of the theory constructed