Variability associated with interpreting drugs within forensic hair analysis: A three-stage interpretation.

Hair analysis is capable of determining both an individual's long-term drug history and a single exposure to a drug, which can be particularly important for corroborating incidents of drug-facilitated crimes. As a source of forensic evidence that may be used in a court of law, it must be credible, impartial and reliable, yet the pathways of drug and metabolite entry into hair are still uncertain. Many variables may influence drug analysis results, most of which are outside of the control of an analyst. An individual's pharmacokinetic and metabolic responses, hair growth rates, drug incorporation routes, axial migration, ethnicity, age and gender, for example, all display interpersonal variability. At present there is little standardization of the analytical processes involved with hair analysis. Both false positives and negative results for drugs are frequently encountered, regardless of whether a person has consumed a drug or not. In this regard, we have categorized these variables and proposed a three-stage analytical approach to facilitate forensic toxicologists, hair analysis experts, judiciaries and service users in the analytical and interpretation process.

Investigating the Opposing Effect of Two Different Green Tea Supplements on Oxidative Stress, Mitochondrial Function and Cell Viability in HepG2 Cells.

Green tea extract (GTE) improves exercise outcomes and reduces obesity. However, case studies indicate contradictory physiology regarding liver function and toxicity. We studied the effect of two different decaffeinated GTE (dGTE) products, from a non-commercial (dGTE1) and commercial (dGTE2) supplier, on hepatocyte function using the human cell model, HepG2. dGTE1 was protective against hydrogen peroxide (H2O2)-induced apoptosis and cell death by attenuating oxidative stress pathways. Conversely, dGTE2 increased cellular and mitochondrial oxidative stress and apoptosis. A bioavailability study with dGTE showed the major catechin in GTE, EGCG, reached 0.263 µg·ml-1. In vitro, at this concentration, EGCG mimicked the protective effect of dGTE1. GC/MS analysis identified steric acid and higher levels of palmitic acid in dGTE2 versus dGTE1 supplements. We demonstrate the significant biological differences between two GTE supplements which may have potential implications for manufacturers and consumers to be aware of the biological effects of supplementation.

The Impact of a Natural Olive-Derived Phytocomplex (OliPhenolia®) on Exercise-Induced Oxidative Stress in Healthy Adults.

The role of natural polyphenols in reducing oxidative stress and/or supporting antioxidant mechanisms, particularly relating to exercise, is of high interest. The aim of this study was to investigate OliPhenolia® (OliP), a biodynamic and organic olive fruit water phytocomplex, rich in hydroxytyrosol (HT), for the first time within an exercise domain. HT bioavailability from OliP was assessed in fifteen healthy volunteers in a randomized, double-blind, placebo controlled cross-over design (age: 30 ± 2 yrs; body mass: 76.7 ± 3.9 kg; height: 1.77 ± 0.02 m), followed by a separate randomized, double-blinded, cohort trial investigating the short-term impact of OliP consumption (2 × 28 mL∙d−1 of OliP or placebo (PL) for 16-days) on markers of oxidative stress in twenty-nine recreationally active participants (42 ± 2 yrs; 71.1 ± 2.1 kg; 1.76 ± 0.02 m). In response to a single 28 mL OliP bolus, plasma HT peaked at 1 h (38.31 ± 4.76 ng∙mL−1), remaining significantly elevated (p < 0.001) until 4 h. Plasma malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH) and HT were assessed at rest and immediately following exercise (50 min at ~75% V˙O2max then 10 min intermittent efforts) and at 1 and 24 h post-exercise, before and after the 16-day supplementation protocol. Plasma HT under resting conditions was not detected pre-intervention, but increased to 6.3 ± 1.6 ng·mL−1 following OliP only (p < 0.001). OliP demonstrated modest antioxidant effects based on reduced SOD activity post-exercise (p = 0.016) and at 24 h (p ≤ 0.046), and increased GSH immediately post-exercise (p = 0.009) compared with PL. No differences were reported for MDA and CAT activity in response to the exercise protocol between conditions. The phenolic compounds within OliP, including HT, may have specific antioxidant benefits supporting acute exercise recovery. Further research is warranted to explore the impact of OliP following longer-term exercise training, and clinical domains pertinent to reduced oxidative stress.

Saccharin and Sucralose Protect the Glomerular Microvasculature In Vitro against VEGF-Induced Permeability.

Diabetic kidney disease (DKD) has become a global health concern, with about 40% of people living with type 1 and type 2 diabetes mellitus developing DKD. Upregulation of vascular endothelial growth factor (VEGF) in the kidney is a significant pathology of DKD associated with increased glomerular vascular permeability. To date, however, current anti-VEGF therapies have demonstrated limited success in treating DKD. Recent studies have shown that artificial sweeteners exhibit anti-VEGF potential. The aim of this study was therefore to assess the effects of aspartame, saccharin, and sucralose on VEGF-induced leak using an in vitro model of the glomerular endothelium. Saccharin and sucralose but not aspartame protected against VEGF-induced permeability. Whilst the sweeteners had no effect on traditional VEGF signalling, GC-MS analysis demonstrated that the sweetener sucralose was not able to enter the glomerular endothelial cell to exert the protective effect. Chemical and molecular inhibition studies demonstrated that sweetener-mediated protection of the glomerular endothelium against VEGF is dependent on the sweet taste receptor, T1R3. These studies demonstrate the potential for sweeteners to exert a protective effect against VEGF-induced increased permeability to maintain a healthy endothelium and protect against vascular leak in the glomerulus in settings of DKD.

The Impact of Decaffeinated Green Tea Extract on Fat Oxidation, Body Composition and Cardio-Metabolic Health in Overweight, Recreationally Active Individuals.

This study investigated the effect of decaffeinated green tea extract (dGTE), with or without antioxidant nutrients, on fat oxidation, body composition and cardio-metabolic health measures in overweight individuals engaged in regular exercise. Twenty-seven participants (20 females, 7 males; body mass: 77.5 ± 10.5 kg; body mass index: 27.4 ± 3.0 kg·m2; peak oxygen uptake (O2peak): 30.2 ± 5.8 mL·kg-1·min-1) were randomly assigned, in a double-blinded manner, either: dGTE (400 mg·d-1 (-)-epigallocatechin-3-gallate (EGCG), n = 9); a novel dGTE+ (400 mg·d-1 EGCG, quercetin (50 mg·d-1) and α-lipoic acid (LA, 150 mg·d-1), n = 9); or placebo (PL, n = 9) for 8 weeks, whilst maintaining standardised, aerobic exercise. Fat oxidation ('FATMAX' and steady state exercise protocols), body composition, cardio-metabolic and blood measures (serum glucose, insulin, leptin, adiponectin, glycerol, free fatty acids, total cholesterol, high [HDL-c] and low-density lipoprotein cholesterol [LDL-c], triglycerides, liver enzymes and bilirubin) were assessed at baseline, week 4 and 8. Following 8 weeks of dGTE+, maximal fat oxidation (MFO) significantly improved from 154.4 ± 20.6 to 224.6 ± 23.2 mg·min-1 (p = 0.009), along with a 22.5% increase in the exercise intensity at which fat oxidation was deemed negligible (FATMIN; 67.6 ± 3.6%O2peak, p = 0.003). Steady state exercise substrate utilisation also improved for dGTE+ only, with respiratory exchange ratio reducing from 0.94 ± 0.01 at week 4, to 0.89 ± 0.01 at week 8 (p = 0.004). This corresponded with a significant increase in the contribution of fat to energy expenditure for dGTE+ from 21.0 ± 4.1% at week 4, to 34.6 ± 4.7% at week 8 (p = 0.006). LDL-c was also lower (normalised fold change of -0.09 ± 0.06) for dGTE+ by week 8 (p = 0.038). No other significant effects were found in any group. Eight weeks of dGTE+ improved MFO and substrate utilisation during exercise, and lowered LDL-c. However, body composition and cardio-metabolic markers in healthy, overweight individuals who maintained regular physical activity were largely unaffected by dGTE.

Prevalence, determinants and knowledge about herbal medicine and non-hospital utilisation in southwest Nigeria: a cross-sectional study.

OBJECTIVES: To examine the prevalence, determinants, safety perceptions, effectiveness and knowledge of herbal medicines (HMs) and reasons for non-hospital utilisation. DESIGN: Cross-sectional study. SETTING: Ekiti state, southwest Nigeria. PARTICIPANTS: A representative sample (n=1600) of adults (18 years or above) currently living in Ekiti state, southwest Nigeria for at least 2 years, at the time of study. RESULTS: The majority of the respondents (85% n=1265) have used HMs in the last 2 years. Across economic classes use, middle income (88.3%) was the highest (p<0.001), suggesting poverty is not a major factor, even with income inequality. Their use was the most common among respondents with a primary level of education (91.4%, p=0.001); and 100% use (p=0.009) of respondents practising African traditional religion; farmers and those 70 years or above. Our study also reveals more men (p<0.001) used HMs (89.9%) than women (78.6%) and effectiveness was a major reason for use (39.6%) followed by affordability (31.9%). Although the majority of the respondents (90%) knew the difference between certified and uncertified HMs, uncertified ones were the most commonly used (37.3%) in the population. CONCLUSION: Although there is a cultural history of HM use within the study population, the choice of use was based on their effectiveness. Therefore, a scientifically valid analysis of this claim within the study population may help achieve a cheaper and affordable healthcare alternative which will be safe. This is important, considering that uncertified HMs were chosen over certified ones, even though a large majority of respondents were aware of differences and likely consequences. This study highlights the need for further investment by the government, individuals and corporate stakeholders in HM research and improvement of conventional healthcare system. This is in addition to public health awareness on the danger of use of uncertified herbal products.

The effect of paint type on the development of latent fingermarks on walls.

Despite recent advances in DNA technology, fingermark evidence remains a fundamental method of ascertaining an individual's identity. Latent fingermarks are the commonest type of fingermark encountered at crime scenes. The Fingermark Visualisation Manual provides crime scene practitioner's with sequential information regarding which enhancement processes are best suited for a range of deposition surfaces (Bandey et al., 2014) [1]. However, there are still many surfaces, such as painted walls where more knowledge is required regarding which development techniques provide optimum results. In this study, four paint types were tested (matt, silk, bathroom and eggshell). Fingermarks were deposited on painted simulated walls and aged for 1 day, 1 week and 1 month. Fingermarks were developed by three processes highlighted as the most frequently used by practitioners (magnetic granular powder, magneta flake powder and ninhydrin). The results showed that overall black magnetic granular powder outperformed both magneta flake powder and ninhydrin on all paint types. This contradicts current UK guidelines for enhancement of fingermarks on matt painted walls, as black magnetic granular powder is not a recommended process at present. SEM and SEM-EDX analysis showed distinct differences between matt paint and the three non-matt paints tested, which provides an explanation for the results obtained.

The detection of drugs of abuse and pharmaceuticals in drinking water using solid-phase extraction and liquid chromatography-mass spectrometry.

Pharmaceuticals and drugs of abuse including novel psychoactive substances (NPS) are emerging as newer contaminants in the aquatic environment. The presence of such pollutants has implications on the environment as well as public health and therefore their identification is important when monitoring water quality. This research presents a new method for the simultaneous detection of 20 drugs of abuse and pharmaceuticals in drinking water, including 15 NPS, three traditional illicit drugs and two antidepressants. The developed method is based on the use of solid-phase extraction (SPE) followed by liquid chromatography-mass spectrometry (LC-MS). The SPE recoveries for the majority of target analytes ranged between 62 and 107%. The method detection and quantification limits ranged between 0.01 and 1.09 ng/L and 0.02-3.64 ng/L respectively. Both instrumental and method precisions resulted in relative standard deviations <15.04%, with an accuracy of < ±8.66%. The results show that LC-MS can be an alternative to the more popular technique of liquid chromatography-tandem mass spectrometry for the analysis of drugs of abuse and pharmaceuticals in drinking water. This newly developed simultaneous detection method has been applied to drinking water collected from the East Anglia region of the UK. Citalopram, cocaine, fluoxetine, ketamine, mephedrone, methamphetamine and methylone were detected at the range of 0.14 and 2.81 ng/L. This is the first time that the two NPS mephedrone and methylone, have been detected in UK drinking water.

A multifactorial critical appraisal of substances found in drug facilitated sexual assault cases.

Drug-facilitated sexual assault (DFSA) is a sexual act in which the victim is unable to give or rescind consent due to intoxication with alcohol and/or drugs that have been self-administered (opportunistic DFSA) or covertly administered by the perpetrator (predatory DFSA). The drugs that are most commonly associated with DFSA are flunitrazepam and gamma-hydroxybutyric acid (GHB). They cause sedation and amnesia, are readily dissolved in beverages and are rapidly eliminated from the system. However, drugs such as amphetamine and cocaine, which are central nervous system (CNS) stimulants, have also been encountered in DFSA cases. This paper critically evaluates trend data from cohort studies, identifying drugs that have been detected in DFSA cases and reports on the differences in drugs used between opportunistic and predatory DFSA. This is the first time that a critical multifactorial review of drugs used in DFSA has been conducted. The pharmacology of each identified group of drugs is presented, showing why these compounds are of interest and used in the perpetration of DFSA. Furthermore, the pharmacology and mechanisms of action are described to explain how the drugs cause their effects. It is also apparent from this study that if meaningful data is to be exchanged between law enforcement agencies then it is necessary to agree on protocols for the collection of evidence and the drugs for which analysis should be performed and indeed on the analytical methods used.

Amphetamine binding to synthetic melanin and scatchard analysis of binding data.

Previous research into drug-hair binding shows that hair color affects drug-hair binding. There are no structural disparities in hair of different colors other than the type and content of melanin present. For this reason, this investigation focuses on synthetic eumelanin as a site for drug interaction using amphetamine as the candidate drug. The binding study was carried out at room temperature. The interaction between synthetic eumelanin and amphetamine was monitored using UV-Vis spectrophotometry at 257.2 nm. As the molecular weight of melanin is unknown, the number of binding sites could not be calculated directly. Hence the ratio of the number of mumoles of drug bound and the dry weight of melanin in mug was considered. Equilibrium was reached when approximately 32% of the drug was bound to melanin. Hence this study proves that amphetamine binds to synthetic eumelanin in vitro. Data interpretation using Scatchard analysis yielded a curvilinear plot with upward concavity indicating multiple binding sites on melanin and negative cooperativity.

Drug facilitated sexual assault: detection and stability of benzodiazepines in spiked drinks using gas chromatography-mass spectrometry.

Benzodiazepines are detected in a significant number of drug facilitated sexual assaults (DFSA). Whilst blood and urine from the victim are routinely analysed, due to the delay in reporting DFSA cases and the short half lives of most of these drugs in blood and urine, drug detection in such samples is problematic. Consideration of the drinks involved and analysis for drugs may start to address this. Here we have reconstructed the 'spiking' of three benzodiazepines (diazepam, flunitrazepam and temazepam) into five drinks, an alcopop (flavoured alcoholic drink), a beer, a white wine, a spirit, and a fruit based non-alcoholic drink (J2O) chosen as representative of those drinks commonly used by women in 16-24 year old age group. Using a validated GC-MS method for the simultaneous detection of these drugs in the drinks we have studied the storage stability of the benzodiazepines under two different storage conditions, uncontrolled room temperature and refrigerator (4°C) over a 25 day period. All drugs could be detected in all beverages over this time period. Diazepam was found to be stable in all of the beverages, except the J2O, under both storage conditions. Flunitrazepam and temazepam were found not to be stable but were detectable (97% loss of temazepam and 39% loss of flunitrazepam from J2O). The recommendations from this study are that there should be a policy change and that drinks thought to be involved in DFSA cases should be collected and analysed wherever possible to support other evidence types.