Jon E. Sprague

The complementary and divergent roles of uncoupling proteins 1 and 3 in thermoregulation.

Both uncoupling protein 1 (UCP1) and UCP3 are important for mammalian thermoregulation. UCP1 and UCP3 in brown adipose tissue mediate early and late phases of sympathomimetic thermogenesis, respectively. Lipopolysaccharide thermogenesis requires skeletal muscle UCP3 but not UCP1. Acute noradrenaline‐induced hyperthermia requires UCP1 but not UCP3. Loss of both UCP1 and UCP3 accelerate the loss of body temperature compared to UCP1KO alone during acute cold exposure.

Impact of common clandestine structural modifications on synthetic cathinone “bath salt” pharmacokinetics

ABSTRACT Since 2009, the synthetic cathinones (“bath salts”) have risen in popularity as drugs of abuse. However, there are a paucity of studies that have determined the impact of functional group modifications in the synthetic cathinone chemical structures on plasma and central nervous system (CNS) pharmacokinetics. In the present study, we investigated the in vivo plasma and CNS pharmacokinetics of three synthetic cathinones whose structures differ by lengthening of the &agr;‐alkyl chain: methylone (‐CH3), butylone (‐CH2CH3), and pentylone (‐CH2CH2CH3). Male Sprague‐Dawley rats were treated with a 20 mg/kg subcutaneous dose of the individual synthetic cathinone. Blood samples were obtained at specific times from a jugular vein cannula over an 8 hour period. Over a separate three‐hour period, CNS samples were obtained using a microdialysis cannula surgically implanted into the lateral ventricle. In the plasma, pentylone, with the longest &agr;‐alkyl chain, displayed the highest Cmax and AUC0–∞, and the longest t1/2. Decreasing the &agr;‐alkyl chain length as in butylone and methylone significantly decreased the Cmax, AUC0–∞, and t1/2. The plasma pharmacokinetic values are consistent with the greater lipophilicity associated with &agr;‐alkyl side chain lengthening. Conversely, in the CNS, methylone and butylone displayed higher Cmax and AUC0–∞ values than pentylone. These contrary findings in the CNS and plasma demonstrate that lengthening of the &agr;‐alkyl chain of methylone, butylone, and pentylone yields differential pharmacokinetic properties in the CNS as compared to the plasma. HIGHLIGHTSAdding &agr;‐methyl groups increase synthetic cathinone plasma Cmax, AUC0–∞, t1/2.Adding &agr;‐methyl groups decrease synthetic cathinone CNS Cmax, AUC0–∞, t1/2.The CNS has greater exposure to the synthetic cathinones as compared to the plasma.

Statistical Modeling of the Case Information From the Ohio Attorney General's Sexual Assault Kit Testing Initiative

The Ohio Attorney Generals Sexual Assault Kit (SAK) Testing Initiative has resulted in nearly 14,000 kits being processed since the initiation of the project in 2012. A logistic regression model was fit to the data from 2500 SAKs in order to determine the probability of obtaining at least one Combined DNA Index System (CODIS) eligible DNA profile based on a number of predictor variables. The probability of obtaining at least one CODIS eligible DNA profile from an SAK varied as a function of (i) days to kit collection following a sexual assault; (ii) years to kit submission to the laboratory for testing following kit collection; (iii) the age of the victim; and (iv) the occurrence of victim‐reported consensual sex around the time of the assault and/or kit collection. These findings demonstrate the utility of the statistical modeling of data obtained from the “forklift” testing approach of sexual assault kits.

Potentiation of Ecstasy-induced hyperthermia and FAT/CD36 expression in chronically exercised animals

ABSTRACT Fatal hyperthermia as a result of 3,4-methylenedioxymethamphetamine (MDMA) use involves non-esterified free fatty acids (NEFA) and the activation of mitochondrial uncoupling proteins (UCP). NEFA gain access into skeletal muscle via specific transport proteins, including fatty acid translocase (FAT/CD36). FAT/CD36 expression is known to increase following chronic exercise. Previous studies have demonstrated the essential role of NEFA and UCP3 in MDMA-induced hyperthermia. The aims of the present study were to use a chronic exercise model (swimming for two consecutive hours per day, five days per wk for six wk) to increase FAT/CD36 expression in order to: 1) determine the contribution of FAT/CD36 in MDMA (20 mg/kg, s.c.)-mediated hyperthermia; and 2) examine the effects of the FAT/CD36 inhibitor, SSO (sulfo-N-succinimidyl oleate), on MDMA-induced hyperthermia in chronic exercise and sedentary control rats. MDMA administration resulted in hyperthermia in both sedentary and chronic exercise animals. However, MDMA-induced hyperthermia was significantly potentiated in the chronic exercise animals compared to sedentary animals. Additionally, chronic exercise significantly reduced body weight, increased FAT/CD36 protein expression levels and reduced plasma NEFA levels. The FAT/CD36 inhibitor, SSO (40 mg/kg, ip), significantly attenuated the hyperthermia mediated by MDMA in chronic exercised but not sedentary animals. Plasma NEFA levels were elevated in sedentary and exercised animals treated with SSO prior to MDMA suggesting attenuation of NEFA uptake into skeletal muscle. Chronic exercise did not alter skeletal muscle UCP3 protein expression levels. In conclusion, chronic exercise potentiates MDMA-mediated hyperthermia in a FAT/CD36 dependent fashion.

Technical note: The effects of Bluestar® and luminol when used in conjunction with tetramethylbenzidine or phenolphthalein

There are numerous presumptive tests available in the forensic science field to help identify the presence of blood. While many articles are available on the effects of Bluestar(®) and luminol and potential interactions with subsequent DNA identification, the research field falls short in identifying the effects these two presumptive tests may have on subsequent presumptive tests used to help identify blood. To rectify this ongoing issue in the forensic science field, the chemiluminescence methods of Bluestar(®) and luminol for the detection of blood at a crime scene were tested for their effects when used in conjunction with tetramethylbenzidine (TMB) or phenolphthalein (PT) at the forensic science laboratory. Six different substrates (untreated wood, pressure treated wood, ceramic tile, shag carpet, cement block, and cotton clothing) were stained with varying dilutions (range 1:1 to 1:100,000) of blood. Neither luminol nor Bluestar(®) affect the results of PT or TMB tests at blood dilutions equal to or less than 1:100. However, interactions did occur between agents and substrates with blood dilutions 1:1000 or greater. Bluestar(®) was the only presumptive test that can detect blood dilutions of 1:100,000 on some substrates and luminol was inclusive on pressure treated wood. These findings suggests that forensic science laboratory personal need to know and understand the details of how the blood was detected by the crime scene investigator and the substrate on which the blood was obtained from for their preparation of presumptive blood testing with PT or TMB.

Comparison of Decision Tree and Logistic Regression Models for Utilization in Sexual Assault Kit Processing

To combat the influx of sexual assault kits (SAKs) that need to be tested, an exploration of data from Ohios SAK Testing Initiative was carried out to identify variables that impact whether a SAK contains a probative DNA profile that is eligible for the Combined DNA Index System (CODIS) database. A validation study was completed to confirm the existence of variable relationships from the initial examination of data; new and modified statistical models were introduced to improve the predictive accuracy to determine if a SAK will contain at least one CODIS eligible DNA profile. Descriptive statistics from the validation data set confirmed conclusions about the effects of days between the assault and kit collection, the age of the victim, and consensual sex around the time of the kit collection for obtaining CODIS eligibility of DNA. The decision tree was selected as the best model.

Letter to the Editor-Elbow Grease and OxiClean™ for Cleaning Fentanyl- and Acetylfentanyl-contaminated Surfaces

Sir, Fentanyl is a schedule II prescription drug under the Controlled Substances Act (1). Fentanyl is a powerful synthetic opioid analgesic 50–100 times more powerful than morphine. Fentanyl is indicated for the treatment of severe pain (2). Recently, fentanyl and synthetic opioids have been increasingly abused, with the death rate associated with opioids and synthetic opioids rising 72.2% from 2014 to 2015 (3). This rapid increase in toxicological events is not only putting drug users at risk, but also first responders and medical professionals treating overdose victims (4,5). There is, therefore, a critical need to identify effective and safe methods for cleaning fentanyl and synthetic forms of fentanyl from contaminated surfaces. Here, we examine the effects of OxiClean Versatile Stain Remover or water on cleaning a contaminated surface. The present study was designed to mimic a typical chemical spill in a laboratory or in a law enforcement setting. Fentanyl and acetylfentanyl were purchased from Cayman Chemical (Ann Arbor, MI). A 1% (w/v) solution of 5 g of OxiClean Versatile Stain Remover in 500 mL of tap water was made. This specific OxiClean product contains sodium percarbonate (SPC), which has previously been shown to oxidize fentanyl in a laboratory setting (6). The solution was mixed in a spray bottle with gradients for easy liquid measurements. Proper safety equipment was worn throughout the duration of the experiment, including doubled nitrile or latex gloves changed every 10 min, disposable laboratory coats (preferably those that close in the back), particulate masks such as The National Institute for Occupational Safety and Health (NIOSH) recommended N-100, P-100, or R-100 particulate filtering facepiece respirator, and safety goggles (7). All equipment except the goggles were placed in biohazardous waste containers at the conclusion of the study. While this would be considered as a toxic substance and ideally be discarded into a toxic waste container, the majority of law enforcement agencies do not have this type of waste treatment ability. Most do, however, have biohazardous waste containers due to frequent contact with bodily fluids found on evidence. The Ohio Environmental Protection Agency has multiple approved methods for disposal of biohazardous material, including incineration, chemical, or heat treatments with or without chemical assistance (8). All of these methods should be sufficient in neutralizing fentanyl. The experiment was performed on two benchtops at the Ohio Attorney General’s Bureau of Criminal Investigation in Bowling Green, OH. One benchtop was used solely for fentanyl, and the other was used solely for acetylfentanyl. The benchtops were swabbed with a cotton swab soaked in methanol (Sigma-Aldrich,

University students’ attributions for abstinence from synthetic cannabinoids and synthetic cathinones

ABSTRACT Background: Synthetic cannabinoids and synthetic cathinones are two increasingly available and potentially dangerous classes of substances. Objective: We designed this study to test whether university students rated the influence of different types of reasons for abstaining differently as a function of type of drug (synthetic cannabinoids vs. synthetic cathinones) and gender (male vs. female). Method: Using a web-based survey, 176 male and 437 female undergraduate university students rated the degree to which each of 42 reasons for not taking drugs influenced their abstinence from those two classes of substances. Results: Exploratory factor analyses suggested four subscales of reasons applicable to both substances: (1) psychological and behavioral impairment, (2) somatic and physiological concerns, (3) social approval and self-image concerns, and (4) insufficient knowledge and limited access. Both men and women rated all four subscales of reasons as having more influence on their abstinence from synthetic cathinones than synthetic cannabinoids, and women rated each subscale except somatic and physiological concerns as having more influence than did men. Conclusions: Although there were main effects for type of drug, because students typically reported the same types of reasons as being more or less influential for both classes of substances, prevention interventions could focus simultaneously on reasons to avoid or delay use of both types of substances.

The α- And β-adrenergic antagonist controversy with sympathomimetic agents

We read with great interest the Letter to the Editor by Richards et al., published in response to our article on synthetic cathinones, or “bath salts” (1,2). Although we applaud the authors attempt to shed some light on the controversial use of selective b-adrenergic receptor antagonists as treatment for toxicity produced by sympathomimetic agents, such as the synthetic cathinones, several statement and claims made by Richards et al., require further clarification. They state that they . “routinely utilize intravenous labetalol, a mixed aand b-blocker, in the treatment of methamphetamine, cocaine and ‘bath salt’ patients, with excellent results.” Important, and underemphasized by these authors, is the fact that labetalol possesses a-adrenergic receptor antagonistic properties. Additionally, we outlined the important role activation of aand b-adrenergic receptors play in the cardiovascular and hyperthermic responses seen to sympathomimetic agents, such as the synthetic cathinones. Previously, we had shown in a preclinical model that b-adrenergic receptor antagonists devoid of a-adrenergic receptor antagonistic properties were not effective in attenuating the hyperthermia induced by the sympathomimetic agent, 3,4-methylenedioxymethamphetamine (MDMA) (3). In this same study, we demonstrated that carvedilol, an a1and b1,2,3-adrenergic receptor antagonist, could attenuate MDMA-induced hyperthermia and rhabdomyolysis (3). Subsequent to our findings with carvedilol, Hysek et al. demonstrated in healthy human subjects that carvedilol could reverse MDMA-mediated increases in blood pressure, heart rate, and body temperature (4). Hysek et al. had previously demonstrated in healthy human subjects that the nonselective b-adrenergic receptor antagonist (devoid of a-adrenergic receptor activity), pindolol, inhibited MDMA-induced increases in heart rate, but was ineffective at preventing the increase in mean arterial blood pressure (5).