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Ethanol stability in antemortem blood stored under various conditions has been widely studied. Most such studies have somewhat limited sample size (

The High Drinking in the Dark mouse lines (HDID-1 and HDID-2) were selectively bred to achieve high blood ethanol concentrations (BECs) in the Drinking in the Dark (DID) task, a widely used model of binge-like intake of 20% ethanol. There are several components that differentiate DID from other animal models of ethanol intake: time of day of testing, length of ethanol access, single-bottle access, and individual housing. Here, we sought to determine how some of these individual factors contribute to the high ethanol intake observed in HDID mice. HDID-1, HDID-2, and non-selected HS/NPT mice were tested in a series of DID experiments where one of the following factors was manipulated: length of ethanol access, fluid choice, number of ethanol bottles, and housing condition. We observed that 1) HDID mice achieve intoxicating BECs in DID, even when they are group-housed; 2) HDID mice continue to show elevated ethanol intake relative to HS/NPT mice during an extended access session, but this is most apparent during the first 4 h of access; and 3) offering a water choice during DID prevents elevated intake in the HDID-1 mice, but not necessarily in HDID-2 mice. Together, these results suggest that the lack of choice in the DID paradigm, together with the length of ethanol access, are important factors contributing to elevated ethanol intake in the HDID mice. These results further suggest important differences between the HDID lines in response to procedural manipulations of housing condition and ethanol bottle number in the DID paradigm, highlighting the distinct characteristics that each of these lines possess, despite being selectively bred for the same phenotype.

Hemolysis, a common occurrence in blood collected for chemical analysis, has been reported to affect analytical test results for some analytes depending upon the material tested and the analytical technique employed. The potential for hemolysis to impact blood ethanol determinations using headspace gas chromatography of samples diluted with an internal standard was investigated. A sample of non-hemolyzed blood and a matched sample of hemolyzed blood were both analyzed thirty times for ethanol concentration using headspace gas chromatography. The mean ethanol concentration measured for the non-hemolyzed samples was 0.0639 g/dl. The mean ethanol concentration measured for the hemolyzed samples was 0.0642 g/dl. The calculated t value, 1.897, was less than the critical t value, 2.002, at a 0.05 level of significance. There was no measured statistical difference detected between the mean blood ethanol concentration determined for a hemolyzed whole blood sample and a non-hemolyzed whole blood sample.

Ethanol can have both an aversive and rewarding effect, which may have a significant relationship to its individual preference. So far, the reasons for the high and low ethanol preference in the WHP (Warsaw High Preferring) and WLP (Warsaw Low Preferring) lines have not been found. WHP rats spontaneously drink over 5 g/kg/day of ethanol, while WLP rats drink under 2 g/kg/day. The purpose of the work was to study the sensitivity of WHP and WLP rats to the aversive effects of ethanol at doses of 1.5 g/kg and 2.0 g/kg in the conditioned taste aversion (CTA) procedure. Lower doses (0.5 and 1.0 g/kg, i.p. [intraperitoneally]) were tested earlier and only 1.0 g/kg produced a slight aversion in WLP rats. The secondary aim was to check the additional potential factors (blood ethanol concentration, pain sensitivity, anxiety-related behavior, learning, and memory) that may constitute an important differentiating feature of the WHP and WLP lines. For this purpose, the following tests were conducted: blood ethanol concentration, novel object recognition (NOR), flinch-jump, hot-plate, and elevated plus maze (EPM). The 1.5 g/kg i.p. dose of ethanol caused the development of an aversion only in WLP rats and the aversion extinguished in the post-conditioning phase. The 2.0 g/kg i.p. dose of ethanol resulted in the development of an aversion in both the tested groups, with the aversion being maintained throughout the whole post-conditioning period only in the WLP rats. There were no differences between the lines in terms of the blood ethanol concentration and the EPM tests. WHP rats had a higher pain sensitivity compared to WLP rats in flinch-jump and hot-plate tests. WLP rats showed a shorter exploration time for both objects compared to WHP in the NOR test. In conclusion, WHP and WLP rats differ in sensitivity to the aversive effects of ethanol. This difference may partially explain their opposite ethanol preference.

Variability in the blood-breath ratio (BBR) of alcohol is important, because it relates a measurement of the blood-alcohol concentration (BAC) with the co-existing breath-alcohol concentration (BrAC). The BBR is also used to establish the statutory BrAC limit for driving from the existing statutory BAC limits in different countries. Thein-vivoBBR depends on a host of analytical, sampling and physiological factors, including subject demographics, time after end of drinking (rising or falling BAC), the nature of the blood draw (whether venous or arterial) and the subjects breathing pattern prior to exhalation into the breath analyzer. The results from a controlled drinking study involving healthy volunteers (85 men and 15 women) from three ethnic groups (Caucasians, Hispanics and African Americans) were used to evaluate various factors influencing the BBR. Ethanol in breath was determined with a quantitative infrared analyzer (Intoxilyzer 8000) and BAC was determined by headspace gas chromatography (HS-GC). The BAC and BrAC were highly correlated (r = 0.948) and the BBR in the post-absorptive state was 2 382 +/- 119 (mean +/- SD). The BBR did not depend on gender (female: 2 396 +/- 101 and male: 2 380 +/- 123,P > 0.05) nor on racial group (Caucasians 2 398 +/- 124, African Americans 2 344 +/- 119 and Hispanics 2 364 +/- 104,P > 0.05). The BBR was lower in subjects with higher breath- and body-temperatures (P < 0.05) and it also decreased with longer exhalation times into the breath-analyzer (P < 0.001). In the post-absorptive state, none of the 100 subjects had a BBR of less than 2 100:1.

Background Beyond yielding high blood ethanol (EtOH) concentrations (BECs), binge-drinking models allow examination of drinking patterns which may be associated with EtOH's rewarding effects, including front-loading and consummatory successive negative contrast (cSNC), a decrease in intake when only water is available to subjects expecting EtOH. The goals of the current study were to broaden our understanding of these reward-related behaviors during binge EtOH access in high alcohol-preferring (HAP) replicate lines (HAP2 and HAP3) of mice selectively bred to prefer alcohol. We hypothesized that both lines would show evidence of front-loading during binge EtOH access and that we would find a cSNC effect in groups where EtOH was replaced with water, as these results have been shown previously in HAP1 mice. Methods HAP replicate 2 and replicate 3 female and male mice were given 2 hours of EtOH or water access in the home cage for 15 consecutive days using "drinking in the dark" (DID) procedures. Mice received the same fluid (either 20% unsweetened EtOH or water) for the first 14 days. However, on the 15th day, half of the mice from these 2 groups were provided with the opposite assigned fluid (EtOH groups received water and vice versa). Intake was measured in 1-minute bins using specialized sipper tubes, which allowed within-session analyses of binge-drinking patterns. Results EtOH front-loading was observed in both replicates. HAP3 mice displayed front-loading on the first day of EtOH access, whereas front-loading developed following alcohol experience in HAP2 mice, which may suggest differences in initial sensitivity to EtOH reward. Consummatory SNC, which manifests as lower water intake in mice expecting EtOH as compared to mice expecting water, was observed in both replicates. Conclusions These findings increase confidence that defined changes in home cage consummatory behavior are driven by the incentive value of EtOH. The presence of cSNC across HAP replicates indicates that this reaction to loss of reward is genetically mediated, which suggests that there is a biological mechanism that might be targeted.

The stability of ethanol in antemortem blood stored under various conditions has been widely studied. Antemortem blood samples stored at refrigerated temperature, at room temperature, and at elevated temperatures tend to decrease in ethanol concentration with storage. It appears that the stability of ethanol in blood exposed to temperatures greater than 38°C has not been evaluated. The case presented here involves comparison of breath test results with subsequent analysis of blood drawn at the time of breath testing. However, the blood tubes were in a refrigerator fire followed by refrigerated storage for 5 months prior to analysis by headspace gas chromatography. The subject's breath was tested twice using an Intoxilyzer 8000. The subject's blood was tested in duplicate using an Agilent headspace gas chromatograph. The measured breath ethanol concentration was 0.103 g/210 L and 0.092 g/210 L. The measured blood ethanol concentration was 0.0932 g/dL for both samples analyzed. Although the mean blood test result was slightly lower than the mean breath test result, the mean breath test result was within the estimated uncertainty of the mean blood test result. Even under the extreme conditions of the blood kit being in a refrigerator fire, the measured blood ethanol content agreed well with the paired breath ethanol test.

The correlation between the rising consumption of new psychoactive drugs (NPS), including that of cathinones, and the occurrence of death has not been sufficiently backed up with published analytic data. In fact, the identification of cathinones in human biological samples remains difficult mainly due to the diversity of these substances and their high turnover. In this context, this manuscript aims at documenting a fatal case of a 39-year-old man: autopsy findings consisted in unspecific asphyxic syndrome. Blood ethanol concentration determination and toxicological screenings were performed using gas chromatography with flame ionization detection, liquid chromatography with diode array detection and gas chromatography with mass spectrometry detection, respectively. Liquid chromatography with high-resolution mass spectrometry detection allowed the confirmation of the presence of NPS and the subsequent metabolic study. The analyses have shown the presence of ethanol, tetrahydrocannabinol and two cathinones, 4′-methyl-α-pyrrolidinohexanophenone (MPHP) and N-ethyl-4′-methylnorpentedrone (4-MEAP). MPHP/4-MEAP concentrations were 47/1.6, 97/3.5 and 2380/49,700 µg/L in femoral blood, cardiac blood and urine, respectively. The in vitro metabolic study has highlighted the presence of five metabolites derived from MPHP and three from 4-MEAP but only two metabolites of these products have been detected in biological samples. The 4′-carboxy-PHP, one of the metabolites of MPHP, was detected in every biological sample with higher chromatographic signals than MPHP itself. The number of fatalities related to cathinones use is expected to increase in the coming years. This manuscript reports useful analytical data about MPHP, one of its metabolites (4′-carboxy-PHP) and 4-MEAP.

In clinical laboratories, a common practice used to verify tests prior to reporting is repeat testing. Our objective was to evaluate the differences between the results of blood ethanol concentration (BEC) test repetitions and report on the role of repeat testing to prevent reporting of incorrect results.We conducted a retrospective study of data retrieved from the Bursa Yuksek Ihtisas Training and Research Hospital's document management system by calculating the percentage change between repeated BEC test runs. To assess for clinical relevance, the bias between two results from the same sample was compared using the 1988 Clinical Laboratory Improvement Amendments' (CLIA) proficiency testing allowable total error (TEa) limits.From a total of 1,627 BEC tests performed between January 2017 and January 2018, 70% (1,133) were repeat tested. Of these, 830 resulted in BECs between 0-5 mmol/L, of which 237 (28.5%) were above the 25% acceptable TEa. Two hundred seventy-six BEC test results were greater than14 mmol/L, and there was a good consensus between the initial and repeat test results (99%). In this group, the mean bias was 0.0% (95%, CI = -9.8-9.8%). However, three of the repeat test results were considered significantly different. There were two discordant results in the 5-14 mmol/L ethanol level, and the mean bias was 2.1% (95%, CI = -15.0-19.1%).The majority of the repeated BEC test values were the same as the baseline value; therefore, there may be limited benefit in continuing such frequent repeated analyses.U kliničkim laboratorijama uobičajena je praksa da se izvrši evaluacija testa koji će biti korišćen. Nama je bio cilj da se izvrši evaluacija testa radi utvrđivanja razlika između rezultata pri određivanju koncentracije etanola u krvi (BESC) ponovljenim određivanjem i da iste objavimo kako bi se izbeglo izdavanje netačnih rezultata.Izučavanja su rađena u Bursa Yuksek Training i Research Hospital primenom menadžment sistema radi izračunavanja procenta odstupanja u ponovljenim određivanjima primenom BEC testa. Odgovarajuća klinička procena između rezultata dobijenih u istim uzorcima upore|ivana je primenom 1988 Clinical Laboratory Improvement Amendments (CLIA) testa koji omogućava izračunavanje granica ukupne greške (TEa).Od ukupno 1 627 BEC testova izvedenih između januara 2017 i januara 2018, 70% (1 133) su bili ponovljeni testovi. Od ovih, 830 BEC rezultata bili su između 0–5 mmol/L, a samo je 237 (28,5%) bilo iznad 25% prihvatljive TEa. Dvesta sedamdeset šest rezultata BEC testova bilo je veće od14 mmol/L, i dobijen je dobar konsenzus izmeđuinicijalnih i ponovljenih rezultata testova (99%). U ovoj grupi, srednje odstupanje je bilo 0,0% (95%, CI = -9,8–9,8%). Međutim, tri od ponovljenih rezultata su se značajno razlikovala. Postojala su dva neslaganja između rezultata kod nivoa etanola 5–14 mmol/L, pa je srednje odstupanje bilo 2,1% (95%, CI = -15,0–19,1%).Većina vrednosti ponovljenih BEC testova bila je ista kao i osnovna vrednost, što znači da nema opravdanja da se često vrednosti testova ponavljaju.

This study assessed the ethanol and methanol contents of homemade spirit (Kachasu) sold in Blantyre, Malawi. The likelihood of ethanol and methanol toxicity, respectively, was determined through Monte Carlo simulations using reported Kachasu intake volumes of 21 consumers and the determined methanol and ethanol contents. Ethanol concentration, in samples from 20 different distillers, ranged from 11 to 55% v/v. Methanol was detected in 10 of the 20 samples (0.01–0.28% v/v). The likely mean ethanol intake of drinkers in Blantyre was found to be 214 ± 93 mL per day (90% CI, 68.9–373.4 mL), and mean methanol intake was 0.44 ± 0.37 mL (90% CI, 0.03–1.17 mL). The intake values translated to mean blood ethanol and methanol concentrations of 38 ± 16 mg/mL and 0.05 ± 0.04 mg/mL, respectively. Therefore, the risk of methanol toxicity was considered as negligible. However, there was a high risk of ethanol toxicity. Since production and selling of Kachasu are already illegal in Malawi, enforcement of regulations should be strengthened to reverse the current situation where Kachasu is being distilled and sold openly even within cities. Consumers should also be sensitized about the likely risks associated with consumption of Kachasu in Malawi so that they can make informed choices.

Since the accuracy of headspace gas chromatographic analysis of blood for ethanol concentration has been so well established over the past several decades, it has become commonplace in court proceedings to attack preanalytical handling of the blood samples including the lack of measuring sample temperature prior to sample preparation. The impact on measured ethanol concentration of allowing refrigerated (~4℃) samples varying amounts of time to equilibrate with room temperature, 24, 4, 3, 2, and 1 h, prior to sample preparation was evaluated. Samples were diluted 1:10 with an internal standard using a diluter/dispenser and analyzed using headspace gas chromatography. The mean ethanol concentration measured for the sixteen samples at each of the five equilibration times was 0.153 g/dl. The F-critical from the one-way ANOVA was 2.4937. The calculated F value was 0.4209. Additionally, the effect on measured ethanol concentration of having calibrators at different temperatures than case samples was investigated. Three groups were analyzed: all calibrators, controls, and samples given 24 h to equilibrate with room temperature, all calibrators, controls, and samples prepared immediately after removal from refrigeration, and calibrators sampled immediately after removal from refrigerator with samples and controls allowed 24 h to equilibrate with room temperature. The mean ethanol concentration measured for the thirty blood samples in each of the three groups was 0.197 g/dl. The F-critical from the one-way ANOVA was 3.1013. The calculated F value was 0.0188. Measured ethanol concentrations were insensitive to the variations in preanalytical conditions evaluated in this study.

Animal models of alcohol drinking and dependence are a critical resource for understanding the neurobiological mechanisms and development of more effective treatments for alcohol use disorder (AUD). Because most rat strains do not voluntarily consume large enough quantities of alcohol to adequately model heavy drinking, dependence, and withdrawal-related symptoms, researchers frequently turn to experimenter administered methods to investigate how prolonged and repeated exposure to large quantities of alcohol impacts brain and behavior. Vaporized ethanol is a common method used for chronically subjecting rodents to alcohol and has been widely used to model both binge and dependence-inducing heavy drinking patterns observed in humans. Rodent strain, sex, and age during exposure are all well-known to influence outcomes in experiments utilizing intraperitoneal or intragastric methods of repeated ethanol exposure. Yet, despite its frequent use, the impact of these variables on outcomes associated with ethanol vapor exposure has not been widely investigated. The present study analyzed data generated from over 700 rats across an eight-year period to provide a population-level assessment of variables influencing level of intoxication using vapor exposure. Our findings reveal important differences with respect to strain, sex, and age during ethanol exposure in the relationship between blood ethanol concentration and behavioral signs of intoxication. These data provide valuable scientific and practical insight for laboratories utilizing ethanol vapor exposure paradigms to model AUD in rats.

In forensic toxicology, alternative biological materials are very useful and important, e.g. in the case of lack of basic body fluids. One alternative biological material is cerebrospinal fluid (CSF). The procedures of the collection of biological material during the autopsy are performed in accordance with local, usually national recommendations, which most often require updating. It is very difficult to assess the possibility of using CSF as an alternative biological material for toxicological studies for the presence of drugs, intoxicants, including new psychoactive substances (commonly known as designer drugs), psychotropic substances, and ethyl alcohol, based on current data. Previous research suggests that CSF may be useful in toxicological studies, but these aspects need to be investigated more carefully because studies have collected CSF from different sites and often the results of different authors are not comparable. It would be necessary to prepare guidelines, e.g. the site of CSF collection that may influence the results of quantitative analysis. It would also be necessary to replicate some studies with a different collection site or a more recent analytical technique, e.g. for comparative testing of blood ethanol and cerebrospinal fluid. Cerebrospinal fluid can be a valuable information carrier in the absence of classic biological material from an autopsy. Investigating these aspects in more detail could allow the future use of this alternative material for routine toxicology analyzes in a forensic laboratory.

A 72-year-old male was arrested for impaired driving. He had a history of chronic alcohol1 consumption and limited food intake on the day of his arrest. The Intoxilyzer® 8000C provided the following communication messages during testing: “Interferent Detect”, “Invalid Sample” and then an additional “Interferent Detect.” This information from the Intoxilyzer® 8000C prompted a medical investigation. The individual was taken to the hospital where ketoacidosis was part of the medical diagnosis. Blood samples were collected and analyzed at two hospital laboratories and the Centre of Forensic Sciences. The results from the hospital analyses were serum ethanol concentrations of 172 and 161 mg/100 mL and an acetone concentration of 18 mg/100 mL; the results from the forensic analyses were blood ethanol, acetone, isopropanol and beta-hydroxybutyrate concentrations of 139 mg/100 mL, 21 mg/100 mL

Alcohol use disorder affects millions of people each year. Currently approved pharmacotherapies have limited success in treating this disorder. Evidence suggests that this lack of success is partly due to how these pharmacotherapies are tested in preclinical settings. The vast majority of preclinical studies assessing the effects of pharmacotherapies on alcohol or drug self-administration are done in individually housed animals. However, it is known that alcohol and drug intake are heavily influenced by social settings. Here, we adapted radio frequency tracking technology to determine the effects of oxytocin, a potential therapy for alcohol use disorder, on alcohol consumption in socially housed male and female prairie voles. Voluntary alcohol consumption in these animals resulted in high daily alcohol intakes, blood ethanol concentrations that are considered intoxicating, and central changes in FosB immunoreactivity, indicative of changes in neural activity. Prairie voles that received oxytocin temporarily reduced alcohol consumption but not alcohol preference, compared with control prairie voles regardless whether their cagemates received a similar treatment or not. Our results demonstrate that oxytocin can decrease consummatory behaviors in the presence of peers that are not receiving this treatment, and therefore, its potential use in clinical trials is warranted. Moreover, effectiveness of other pharmacotherapies in preclinical studies can be tested in mixed-treatment socially housed animals similarly to clinical studies in humans.

BackgroundIn rodent studies of ethanol (EtOH) consumption where blood sample collection does not occur, there is often mention of likely BECs based on prior studies. These studies may vary in dose(s) used, age/sex/species, or administration route. Often, intake studies may presume that binge-levels were achieved without knowing that BECs exceeded 80 mg% (binge threshold). In human studies, estimated BECs (eBECs) have been derived using complex formulas that consider EtOH consumption level and the weight and sex of the individual.MethodThree datasets were used to derive eBECs using a conversion factor (CF) that considers gram (g) of EtOH per kilogram (kg) of animal weight and other variables that may influence BECs such as age, sex, dose, route, vehicle, chronicity, and timing post-exposure. Regression analyses were also conducted for each dataset, building regression models with BEC as the response and other variables in the study specific to each dataset as predictor variables.ResultsDataset1 assessed age, sex and post-injection time point. Both CF and regression analyses determined that different CFs should be used for 10- and 30-min post-administration time points. Dataset2 assessed age, dose, vehicle and post-intubation time point. Depending on the post-intubation time point, several CFs were used to derive eBECs. When weight was not used as a regression variable, data across approaches corresponded, with age differences emerging later in elimination phase. In Dataset3 that used BECs from a repeated intake study, chronic exposure influenced CFs, although regression analysis did not yield similar findings.ConclusionsAlthough eBECs can be derived, critical variables vary with subject and test conditions and do not always concur with results of regression analyses. Although, not designed to replace assessment of BECs when sample collection is possible, the CF approach may prove useful when estimating BECs in studies where assessments are not feasible.

Anxiety and aggression are associated with ethanol self-administration, but these behaviors can serve as either risk factors for or consequences of heavy drinking in rodents and humans. Baseline levels of aggressive-like and anxious-like behavior in non-human primates have not yet been characterized in relation to future or prior ethanol intake. The objective of the study was to test the association between temperament at baseline with future ethanol self-administration in late adolescent male (n = 21) and female (n = 11) rhesus monkeys. Shortly after entering the laboratory and before exposure to ethanol, the Human Intruder Test (HIT) and the Novel Object Test (NOT) were used to determine baseline anxious-like and aggressive-like behavior in age-matched male and female rhesus monkeys (Macaca mulatta). The monkeys were induced to drink ethanol 4 % (w/v) using a schedule-induced polydipsia procedure, followed by “open-access” ethanol self-administration in which the monkeys were allowed a choice of water or 4 % ethanol (w/v) for 22 h/day for 52 weeks. Aggressive monkeys self-administered more ethanol and attained higher blood ethanol concentrations (BECs). No significant differences in ethanol intakes or BECs were found between anxious and non-anxious monkeys or between behaviorally inhibited and non-inhibited monkeys. Baseline aggressive behavior positively correlated with ethanol intake and intoxication. Baseline reactive aggression was associated with higher future ethanol intake and intoxication. While significant sex differences in HIT reactivity were observed, the relationship between aggression and ethanol drinking was observed across sex and is not sex-specific.

Background: The stability of ethanol was investigated in blood specimens in glass or plastic evacuated tubes after storage in a refrigerator at 4 °C for up to 12 months. Methods: Sterile blood, from a local hospital, was divided into 50 mL portions and spiked with aqueous ethanol (10% w/v) to give target concentrations of 0.20, 1.00, 2.00 and 3.00 g/L. Ethanol was determined in blood by headspace gas chromatography (HS-GC) with an analytical imprecision of

The aversive action of the pharmacological properties of ethanol was studied in selectively bred Warsaw Alcohol High-Preferring (WHP) and Warsaw Alcohol Low-Preferring (WLP) rats. For this study, a conditioned-taste aversion test was used. Male WHP and WLP rats were submitted to daily 20-min sessions for 5 days, in which a saccharin solution (1.0 g/L) was available (pre-conditioning phase). Next, this drinking was paired with the injection of ethanol (0, 0.5, 1.0 g/kg), intraperitoneally [i.p.] immediately after removal of the saccharin bottle (conditioning phase). Afterward, the choice between the saccharin solution and water was extended for 18 subsequent days for 20-min daily sessions (post-conditioning phase). Both doses of ethanol did not produce an aversion to saccharin in WLP and WHP rats in the conditioning phase. However, injection of the 1.0 g/kg dose of ethanol produced an aversion in WLP rats that was detected by a decrease in saccharin intake at days 1, 3, 7, and 10 of the post-conditioning phase, with a decrease in saccharin preference for 16 days of the post-conditioning phase. Conditioned taste aversion, measured as a decrease in saccharin intake and saccharin preference, was only visible in WHP rats at day 1 and day 3 of the post-conditioning phase. This difference between WLP and WHP rats was apparent despite similar blood ethanol levels in both rat lines following injection of 0.5 and 1.0 g/kg of ethanol. These results may suggest differing levels of aversion to the post-ingestional effects of ethanol between WLP and WHP rats. These differing levels of aversion may contribute to the selected line difference in ethanol preference in WHP and WLP rats.

Binge ethanol consumption has widespread negative consequences for global public health. Rodent models offer exceptional power to explore the neurobiology underlying and affected by binge-like drinking as well as target potential prevention, intervention, and treatment strategies. An important characteristic of these models is their ability to consistently produce pharmacologically-relevant blood ethanol concentration. This review examines the current available rodent models of voluntary, pre-dependent binge-like ethanol consumption and their utility in various research strategies. Studies have demonstrated that a diverse array of neurotransmitters regulate binge-like drinking, resembling some findings from other drinking models. Furthermore, repeated binge-like drinking recruits neuroadaptive mechanisms in mesolimbocortical reward circuitry. New opportunities that these models offer in the current context of mechanistic research are also discussed.

The role of ethanol metabolism system in adaptation of laboratory animals to cold temperatures was shown. Cold stress (1-2°C) modeled in male Wistar rats over 7 weeks significantly modulated endogenous ethanol metabolism and led to reorganization of many physiological systems, which resulted in activation of metabolic processes. Under these conditions, endogenous ethanol was utilized as the most easily and fast metabolized energy substrate, due to which its blood concentration decreased and was replenished at the expense of exogenous ethanol. Normalization of blood ethanol concentration led to better adaptation to cold.

The purpose of this paper is to implement an approximate method for obtaining the solution of a physical model called the blood ethanol concentration system. This model can be expressed by a system of fractional differential equations (FDEs). Here, we will consider two forms of the fractional derivative namely, Caputo (with singular kernel) and Atangana–Baleanu–Caputo (ABC) (with nonsingular kernel). In this work, we use the spectral collocation method based on Chebyshev approximations of the third-kind. This procedure converts the given model to a system of algebraic equations. The implementation of the proposed method to solve fractional models in ABC-sense is the first time. We satisfy the efficiency and the accuracy of the given procedure by evaluating the relative errors. The results show that the implemented technique is an easy and efficient tool to simulate the solution of such models.

Background: Several studies have analyzed the functions of foods and dietary constituents in the dynamics of alcohol metabolism. However, few studies have reported the function of dietary fibers in the dynamics of alcohol metabolism. Objective: We assessed the effects of botanical foods that contain dietary fibers on alcohol metabolism. Methods: The ability of the water-insoluble fraction (WIF) of 18 kinds of botanical foods to maintain 15% (v/v) ethanol solution was examined using easily handled filtration. A simple linear regression analysis was performed to examine the correlation between the filtered volumes and blood ethanol concentration (BEC) in F344 rats 4 h after the ingestion of 4.0 g/kg of ethanol following dosage of 2.5% (w/v) WIF of the experimental botanical foods. Furthermore, the supernatant (6.3 Brix; water-soluble fraction) and precipitate (WIF of tomato), with a strong ethanol-maintaining ability, were obtained and BEC and the residual gastric ethanol in rats were determined 2 h after the administration of 4.0 g/kg of ethanol and the individuals fractions. Results: The filtered volumes of dropped ethanol solutions containing all the botanical foods tested except green peas were decreased compared with the ethanol solution without WIF (control). There was a significant correlation between the filtered volumes and blood ethanol concentration (BEC). There was no significant difference in the residual gastric ethanol between controls and the supernatant group; however, it was increased significantly in the WIF group than in controls or the supernatant group. Consistent with this, BEC reached a similar level in controls and the supernatant group but significantly decreased in the WIF group compared with controls or the supernatant group. Conclusions: These findings suggest that WIFs of botanical foods, which are mostly water-insoluble dietary fibers, possess the ability to absorb ethanol-containing solutions, and this ability correlates strongly with the inhibition of the blood ethanol response, likely by delaying gastric emptying. Key words: Botanical food, tomato, water-insoluble fraction, dietary fiber, blood ethanol concentration

Fatal bear attacks on humans are uncommon with only one reported case in Sweden since 1902. The bear population is, however, growing and the frequency of confrontations is likely to increase. Case I-A 40-year-old hunter and his dog were found dead near a bear's den. Autopsy showed that a large portion of the face, facial skeleton, and anterior portion of the brain was missing. Autopsy of the bear showed two nonfatal gunshot wounds. Case II-A 61-year-old man and his dog were found dead outside a hunting lodge. Autopsy revealed numerous wounds, including partial evisceration of the intestines. The victim's blood ethanol concentration was 0.27%. These cases confirm the presence of risk factors identified by the Scandinavian Brown Bear Research Project, that is, provocation by a dog, encountering an injured bear, and appearing close to its den. An additional possible factor in case II was ethanol intoxication.

Introduction: Measurements of blood ethanol concentrations must be accurate and reliable. The most important factors affecting blood ethanol stability are temperature and storage time. In this study, we aimed to compare ethanol stability in plasma samples at-20 °C for the different storage periods. Materials and methods: Blood samples were collected from intoxicated drivers (N = 80) and initial plasma ethanol concentrations were measured immediately. Plasma samples were then stored at-20 °C and re-assessed after 2, 3, 4, or 5 months of storage. Differences between the initial and stored ethanol concentrations in each group (N = 20) were analyzed using Wilcoxon matched-pairs test. The deviation from the initial concentration was calculated and compared with Clinical Laboratory Improvement Amendments (CLIA’88) Proficiency Testing Limits. Relationships between the initial concentrations and deviations from initial concentrations were analyzed by Spearman’s correlation analysis. For all statistical tests, differences with P values of less than 0.05 were considered statistically significant. Results: Statistically significant differences were observed between the initial and poststorage ethanol concentrations in the overall sample group (P < 0.001). However, for the individual storage duration groups, analytically significant decreases were observed only for samples stored for 5 months, deviations from the initial concentrations exceeded the allowable total error (TEa). Ethanol decreases in the other groups did not exceed the TEa. Conclusion: According to our results, plasma ethanol samples can be kept at-20 °C for up to 3-4 months until re-analysis. However, each laboratory should also establish its own work-flow rules and criterion for reliable ethanol measurement in forensic cases. © by Croatian Society of Medical Biochemistry and Laboratory Medicine.

Background Sudden bath-related deaths occur frequently in Japan, particularly among elderly people. However, the precise mechanism of bath-related death remains uncertain, and effective prevention strategies have not been established. Methods Cases of bath-related deaths (n = 3289) were selected from all cases handled by the Tokyo Medical Examiner's Office from 2009 to 2011 (N = 41 336). The ages and occurrence dates were examined, and major autopsy findings, including toxicological analysis, were evaluated for the autopsied cases (n = 550). Results Most cases occurred in individuals older than 60 years of age during winter. Analysis of autopsy findings revealed water inhalation signs in many cases (n = 435, 79.1%). Circulatory system diseases constituted more than half of the pathological findings regarding factors that may have contributed significantly to death (n = 300, 54.5%), and cardiac lesions were the most common pathological finding (n = 250, 45.5%). However, approximately one-third of the cases exhibited no remarkable pathological findings (n = 198, 36.0%). A quarter of all cases involved blood ethanol levels that exceeded 0.5 mg/mL (n = 140). Conclusions The results suggested that drowning plays an important role in the final process of bath-related death. Circulatory system diseases may be the primary underlying pathology; however, there were variations in the medical histories and pathologies of cases of bath-related death. From a preventive perspective, family members should pay attention to elderly people with circulatory system diseases during bathing, particularly in winter. Additionally, the notion that ill or inebriated individuals should not take baths should be reinforced.

Effects of Opuntia ficus indica (OFI) extracts on ethanol-induced psychomotor alterations were studied using Sprague-Dawley rats orally administered 4 g/kg of ethanol (EtOH group) or distilled water (control group). An OFI-group received OFI extracts (50, 100, and 200 mg/kg) 30 min prior to EtOH administration. Behavioral and hematological tests were evaluated 1, 2, 4, and 8 h after EtOH administration. Electroencephalogram (EEG) assessment was also performed. EtOH significantly (p

Aim: The aim of this study was to obtain reliable data about alcohol consumption in Turkey, to evaluate the reasons for blood ethanol concentration (BEC) analyses in patients admitted to emergency departments, and to evaluate the relationship of BEC with age and time of sampling. Material and Method: A total of 801 patients who were admitted for analyses of BEC was included in the study. The results were classified into three groups according to BEC (50 mg/dl). BEC levels exceeding 10 mg/dl were accepted as ethanol positive (EthPos). The patients were categorized as three groups according to age (40). The cases were classified according to diagnoses: assault, motor vehicle crashes (MVC), injury, suicide, or occupational accident. In addition the patients were grouped according to their time of sampling, daytime or nighttime. Results: MVC was the most common reason for emergency admissions, while assault was the most common cause in EthPos cases. BEC was 100 mg/dL were seen more frequently at night. Assault was the most common cause at night while MVC was most common during the day. EthPos cases were most often found in ages between 18-40. Discussion: MVC constitutes the largest portion of all BEC tests among emergency admissions because individuals involved in any traffic accident are required to be tested for BEC. But assaults are the main causes in EthPos emergency admissions, as it is known that ethanol consumption increases tendencies toward offensive behaviors.

Introduction: The quality of blood ethanol concentration (BEC) determination is important because of its legal ramifications. Measurement uncertainty provides quantitative information about the quality and reliability of test results. In this study, we aim to calculate the measurement uncertainty for the ethanol test in our laboratory measured with a Synchron Systems Ethanol assay kit by employing an enzymatic rate method on the Beckman-Coulter Olympus AU400 auto analyzer (Beckman Coulter Inc, Melville, USA). Materials and methods: The measurement uncertainty values were calculated in accordance to the Nordtest guidelines. All vehicle drivers involved in a traffic accident were retrospectively inspected for the BEC test conducted during July to December 2016 in our emergency laboratory. Results: A 1034 vehicle drivers had their BEC tested. The results for 181 drivers were > 0.50 g/L and reported as positive. The serum ethanol concentration in those showing a positive result was 2.04 ± 1.01 g/L, over four times the legal limit. The median BEC in those showing a negative result was 0.03 (IQR: 0.03) g/L. The expanded uncertainty obtained was 19.74%. When measurement uncertainty values were added to the results of the 1034 drivers who were retrospectively screened, eight vehicle drivers had results with 95% confidence intervals that exceeded the legal limit 0.50 g/L. Conclusions: The BEC test results for vehicle drivers with values close to legal limits should be reported as the obtained ethanol concentration with corresponding measurement uncertainty.

Schedules of intermittent food delivery induce excessive fluid intake, termed schedule-induced polydipsia (SIP), and hypothalamic-pituitary-adrenal (HPA) axis activation is important for the expression and maintenance of this adjunctive behavior. Previous work has focused on examining the relationship between water intake and plasma corticosterone (CORT) in rats at a single or a limited range of fixed time (FT) intervals. However, little remains known regarding SIP and the corresponding stress response (1) across the bitonic function that epitomizes adjunctive behavior, (2) when ethanol is the available fluid, and (3) when a species other than rat or multiple strains are studied. Here we report the findings from ethanol-preferring C57BL/6J (B6) and non-preferring DBA/2J (D2) mice serially exposed to progressively larger FT intervals (0 → 60 min) and given access to either water or a 5% (v/v) ethanol solution. Following 2 weeks of experience with each schedule, blood samples were collected at the conclusion of the last 60-min session to evaluate CORT and the blood ethanol concentration (BEC) achieved. While both strains exhibited a bitonic function of ethanol intake and BEC that peaked at or near a 5-min interval, only D2 mice showed a similar response with water. In contrast, CORT levels rose monotonically with incremental increases in the FT interval regardless of the strain examined or fluid type offered, indicating that glucocorticoid release likely reflects the aversive aspects of increasing intervals between reinforcement rather than engagement in adjunctive behavior. These findings also caution against the use of a single intensity stressor to evaluate the relationship between stress and ethanol intake, as the magnitude of stress appears to affect ethanol consumption in a non-linear fashion.

Background Concurrent alcohol use among cocaine abusers is common but the behavioral variables that promote co-abuse are not well understood. The present study examined the effects of intragastric (i.g.) ethanol (EtOH) administration in monkeys responding under a schedule of cocaine reinforcement in which extensive drug seeking was maintained by conditioned stimuli. Methods Four adult male cynomolgus monkeys ( Macaca fascicularis ) were trained to respond under a second-order fixed-interval (FI) 600 s (fixed-ratio (FR) 30:S) schedule of cocaine (0.003–0.56 mg/kg/injection) presentation. Sessions ended after 5 injections or 90 min had elapsed. Different EtOH doses (0.5–2.0 g/kg, i.g.) were administered 30 min before the session, typically on Tuesdays and Fridays. Blood ethanol concentrations (BECs) were also assessed. Pattern of FI responding was assessed by determining quarter-life (QL) values. Results Cocaine self-administration was characterized as an inverted U-shaped function of dose; QL values increased monotonically with dose. EtOH pretreatments dose-dependently decreased self-administration at several cocaine doses in 3 of 4 monkeys. In one animal, EtOH increased low-dose cocaine-maintained responding. For all monkeys, QL values were increased by EtOH when low- and high-cocaine doses were self-administered, suggesting additive effects of EtOH and cocaine. Furthermore, BECs were not altered following cocaine self-administration. Conclusions The reductions in cocaine self-administration and the increases in QL values following EtOH, suggest that EtOH was enhancing cocaine-related conditioned reinforcement. A better understanding of the behavioral mechanisms that mediate the co-abuse of alcohol and cocaine will lead to improved treatments for both drugs.

In the last century, several mathematical models have been developed to calculate blood ethanol concentrations (BAC) from the amount of ingested ethanol and vice versa. The most common one in the field of forensic sciences is Widmark's equation. A drinking experiment with 10 voluntary test persons was performed with a target BAC of 1.2 g/kg estimated using Widmark's equation as well as Watson's factor. The ethanol concentrations in the blood were measured using headspace gas chromatography/flame ionization and additionally with an alcohol dehydrogenase (ADH)-based method. In a healthy 75-year-old man a distinct discrepancy between the intended and the determined blood ethanol concentration was observed. A blood ethanol concentration of 1.83 g/kg was measured and the man showed signs of intoxication. A possible explanation for the discrepancy is a reduction of the total body water content in older people. The incident showed that caution is advised when using the different mathematical models in aged people. When estimating ethanol concentrations, caution is recommended with calculated results due to potential discrepancies between mathematical models and biological systems.

A role of an altered dietary pattern (e.g., a diet rich in sugar) but also alterations at the level of the intestinal barrier have repeatedly been discussed to be involved in the development and progression of nonalcoholic fatty liver disease (NAFLD).To determine if the nutritional intake, intestinal flora, and permeability and the development of NAFLD are related in humans.Ten controls and 20 patients with NAFLD ranging from simple steatosis to steatohepatitis were included in the study. Bacterial overgrowth, orocecal transit time, and intestinal permeability were assessed. Alcohol, endotoxin, and plasminogen activator inhibitor (PAI-) 1 concentration were determined in plasma. Nutritional intake was assessed using a dietary history.Despite no differences in the prevalence of bacterial overgrowth and in the orocecal transit time, intestinal permeability, alcohol, and endotoxin levels in plasma were significantly higher in patients with NAFLD than in controls. Similar results were also found for PAI-1 plasma concentrations. Patients with NAFLD had a significantly higher intake of protein, total carbohydrates, and mono- as well as disaccharides than controls. PAI-1, endotoxin, and ALT plasma levels were positively related to total protein and carbohydrate intake.Taken together, our results indicate that intestinal permeability, endogenous alcohol synthesis, and nutritional intake are markedly altered in patients with NAFLD.