Why do biological risks exist? Can we develop and implement a strategy for thoughtfully approaching future biological risks? This short, working report provides an abstract introduction to the problem of biological risk and outlines how technical and societal approaches should be combined in order to best address the challenge.

Draft public declaration from the Second International Meeting on Synthetic Biology (May 20-22, 2006, Berkeley, CA)

Thesis (Ph. D.)--Lehigh University, 2000. / Includes bibliographical references and vita.

Cannabinoids are compounds that are naturally present in Cannabis sativa L., which interact with cannabinoid receptors in the nervous system, known as CB1 and CB2 receptors. The most abundant and well-known cannabinoid that can be isolated from cannabis is 9-tetrahydrocannabinol (THC). The structure of this compound specifically allows interaction with the CB1 and CB2 receptors, known as cannabimimetic activity. Other compounds have since been produced, inspired by THC, which have been designed to elicit similar pharmacological responses, and therefore are beneficial as analgesics. These compounds are known as synthetic cannabinoids. Synthetic cannabinoids, while potentially useful as therapeutic treatments for pain, are currently also popular as recreational drugs. Herbal products that contain synthetic cannabinoids are sold as “legal highs,” as few of these compounds are illegal according to the Controlled Drugs and Substances Act. These products are prepared by combining synthetic cannabinoids and plant material, and are smoked similar to marijuana. As the legality of many synthetic cannabinoids is quickly decreasing, as evidenced by the March 2011 emergency scheduling of five such compounds, it is becoming increasingly likely that these products will soon become popular exhibits to be submitted to controlled substances laboratories for testing. If a previously smoked product is submitted, there could potentially be effects due to the burning, the presence of the plant or paper substrate, and other synthetic cannabinoids that could directly diminish the facility of analysis. The aim of this thesis was to investigate these effects using four synthetic cannabinoids (AM-2201, JWH-015, HU-211, and RCS-4) and four substrates (tobacco, rolling paper, mint, and rosemary). Results demonstrated diminished peak areas, which are likely due to the introduction of these variables, which include burning the drug of abuse, and spiking the drug of abuse onto various matrices. The trend of lower peak areas further suggests that burning, the presence of plant material, and other cannabinoids potentially all compromise the facility of analyzing synthetic cannabinoid products. The act of burning one synthetic cannabinoid in particular, AM-2201, appeared to greatly decrease the capability to detect the analyte, as did the application of AM-2201 to various substrates. Furthermore, the ability to detect AM-2201 appeared to vary greatly between results obtained from analyzing samples applied to different substrates. Analysis of cannabinoid mixtures demonstrated that GC/MS analysis of different cannabinoids gave various peak areas although the concentrations remained consistent. Peak area ratios of cannabinoid mixtures that were extracted from substrates were found to not differ significantly between the specific substrates studied. This research supports that all of these variables should therefore be considered in regards to analysis of herbal products containing synthetic cannabinoids.

Designer drugs are some of the most commonly abused substances in the world. They are synthesized through slight chemical modifications of existing substances, evading the law while maintaining the desired effects of the pharmaceutical or illicit substance. These drugs are often marketed as “herbal” or “natural,” but are fully synthetic. Due to their constant, rapid emergence, there is a need for a rapid method of identification, both in the field as well as in the laboratory. One group of these designer drugs are synthetic piperazines. Named for the piperazine ring found in their chemical structures, synthetic piperazines are central nervous system stimulants that have the reputation of mimicking the psychoactive effects of the illicit compounds amphetamine and 3, 4-methylenedioxymethampetamine (MDMA). Over the past 10 years, synthetic piperazine cases submitted to forensic laboratories in the United States have greatly increased, including a 30-fold increase between 2007 and 2009 alone. Surface enhanced Raman spectroscopy (SERS) was investigated as a method for the rapid qualitative analysis of synthetic piperazines. SERS is a type of vibrational spectroscopy, which utilizes the interaction of light and matter to elucidate details of the chemical structure of a molecule. SERS combines laser spectroscopy with the optical properties of metallic nanostructures, resulting in strongly enhanced signals from the Raman scattering of light. Each chemical structure will give a unique SERS spectrum and this, coupled with the minimal-to-no sample preparation and the portability of a SERS instrument, makes SERS a strong candidate for the identification of not only synthetic piperazines, but all designer drugs. To evaluate the use of SERS for the qualitative analysis of synthetic piperazines, eight synthetic piperazines were adsorbed onto a SERS substrate. The interaction with the gold nanoparticles enhanced the Raman scattering for all eight of the synthetic piperazines and SERS spectra were obtained. All eight drugs were found to give a robust and reproducible signal, requiring a fewer number of scans, less laser power, and less time for analysis compared with traditional Raman spectroscopy. When compared with traditional Raman spectra, the synthetic piperazines demonstrated sensitivity enhancement factors of up to 10^8 using SERS. A partial least squares-discriminant analysis (PLS-DA) statistical model was built and used to evaluate the analytical sensitivity and specificity of the SERS method. The PLS-DA model helped determine a limit of detection of 10 μg/mL of BZP. All eight synthetic piperazines could be identified by the statistical model below an error rate of 20% when compared to each other- a strong indication of a method with high specificity.Through this research, it has been demonstrated that SERS can be applied efficiently as a qualitative technique for the analysis of synthetic piperazines.

No description available.

Class of 2012 Abstract / Specific Aims: The purpose of this study is to identify the incidence of symptoms associated after exposure to “bath salts,” a term for synthetic cathinones in Arizona. Methods: This is a retrospective chart review of reported exposures to synthetic cathinones to the Arizona Poison and Drug Information Center and the Banner Good Samaritan Poison and Drug Information Center. Main Results: There were 306 cases of synthetic cathinone exposures reviewed and 76.5% were males (n=234) and 23.5% were females (n=72). They were ingested, inhaled, snorted, or injected. The mean age of exposure to synthetic cathinones was 29 years old. The most common symptoms included agitation (48.7%), hallucinations (27.1%), confusion (17.6%), hypertension (21.9%), tachycardia (50.6%), CK elevation (17.3%) and chest pain (9.5%). Less frequent symptoms exhibited in synthetic cathinone abuse included other CNS effects, gastrointestinal symptoms, muscular dysfunction, visual disturbances, and respiratory issues. Conclusions: The symptoms exhibited after exposure to synthetic cathinones were mainly neurologic and cardiovascular. In most cases, symptoms were effectively resolved within 24 to 48 hours after treatment with intravenous fluids and benzodiazepines. In some reports, patients were also given oxygen, anti-emetics, sedatives and anti-psychotic medications. Medical outcomes included major (1.6%), moderate (42.2%) and minor effects (26.1%) while 92 patients were lost to follow-up.

A dark red resin was isolated from the reaction mixture obtained by treatment of quinoline hydrochloride with zinc dust. Indications, from chromatographic studies of the resin, that it contained a number of different components led to extensive investigations of methods of separation. Satisfactory separation of the resin into pure components was finally achieved by a scheme which essentially involved fractional extraction and fractional adsorption-desorption. Three pure components were isolated, and characterized by chemical and physical analyses. Two of the components were suggested as heterocyclic compounds of quinoline probably of polymeric nature. The third component appeared to be an aniline type compound resulting from reductive cleavage of quinoline. Ultra-violet spectra of the isolated pure components supported the deductions made from chemical and other physical evidence. / Science, Faculty of / Chemistry, Department of / Graduate

Forced vibrational experiments and stress relaxation experiments have been performed on acetate rayon, viscose rayon, nylon, and polyethylene over a range of humidities at both 2°C and 25°C. Forced vibrational experiments have also been carried out on acetate rayon, viscose rayon, and raw silk over the temperature range -80°C to 0°C. Interpretation of previous experiments is discussed in terms of both Newtonian viscosity and Eyring viscosity. The apparatus employed is described and a method for calibrating the solenoid of the forced vibrator is outlined. An inverse relation has been found between ηω and the negative slope of the stress relaxation curve, E°. Since the equation ηω = π/(4.606) E° derived on the basis of Newtonian viscous units predicts a direct relation between ηω and the slope E°, it is assumed that the flow in stress relaxation must be non-Newtonian* Preliminary calculations indicate that a more satisfactory relation between ηω and E° may exist if the flow units involved in stress relaxation follow the Eyring law of viscous flow. An outline for determining such a relation is given. The experimental 1y observed increase with decreasing temperature of dynamic modulus, stress relaxation modulus, and energy loss, as measured by ηω, is interpreted in terms of increasing order of chain segments. A maximum found at -55°C in the ηω vs. temperature curve for viscose rayon is attributed to a "freezing in" of segment motions as the result of a second order transition. / Science, Faculty of / Chemistry, Department of / Graduate

Heterologous biosynthetic pathway expression is an essential tool for natural products biochemists. It has provided a powerful methodology for elucidating and characterizing bacterial biosynthetic pathways. In this thesis I will discuss methods to harness biosynthetic pathways for the heterologous production of a monosaccharide natural product, Legionaminic acid (Leg5,7Ac2). This carbohydrate belongs to a family of sugars called nonulosonic acids (nine carbon α-keto acids) and is a 5,7-diamino derivative of sialic acid (Neu5Ac). It is found in cell surface glycoconjugates of bacteria including pathogens such as Helicobacter pylori, Campylobacter jejuni, Acinetobacter baumanii and Legionella pneumophila. Their presence on bacteria has been correlated with virulence in humans by mechanisms that likely involve subversion of the host’s immune system or interactions with host cell surfaces due to its similarity to sialic acid. Further investigation into their role in bacterial physiology and pathogenicity is limited as there are no effective methods to produce sufficient quantities of these carbohydrates. Herein, I harness microbial biosynthetic pathways via metabolic and genetic engineering to produce these complex nonulosonic acids. Leg5,7Ac2 is produced from N-acetylglucosamine using the Escherichia coli strain BRL04, which results in substantial over-production (> 100 mg L-1 of culture). Pure Leg5,7Ac2 could be readily isolated and converted into CMP-activated Leg5,7Ac2 for biochemical applications as well as the phenyl thioglycoside for chemical synthesis applications. A similar strategy was employed to access the related nonulosonic acid pseudaminic acid (Pse5,7Ac2). A biosynthetic pathway for production of Pse5,7Ac2 was constructed from H. pylori and C. jejuni and expressed in E. coli BRL04. Unlike Leg5,7Ac2, Pse5,7Ac2 was produced in low yields (< 20 mg L-1). A number of modifications were made to the biosynthetic constructs in an effort to enhance production levels yet improved titers were not obtained.Additionally, this thesis will look at the development of a new strategy for the heterologous expression of biosynthetic pathways in a number of diverse hosts. I will highlight a flexible in vivo heterologous expression system that was inspired by viral protein packaging, processing and cleavage to produce violacein, a bright purple pigment with anti-tumor properties. A de novo polyprotein design possessing the violacein biosynthetic pathway was shown to work effectively in prokaryotic hosts such as E. coli and S. typhimurium. Expression of the polyprotein design in eukaryotic hosts like mammalian cells and S. cerevisiae were less successful. The ultimate goal of the work presented herein is to highlight the flexibility and powerful nature of synthetic biology for the in vivo production of natural products in addition to contributing to the vast arsenal of techniques and strategies that are currently available to researchers in this field.