Alcohol use disorder (AUD) and hazardous alcohol use are common but underdiagnosed in primary health care (PHC). This study aimed to explore general practitioners' (GPs') experiences and perceptions of using B-Phosphatidylethanol (PEth), a specific quantitative biomarker for alcohol use, in their clinical work with patient consultations and treatment follow-up in Swedish PHC.

Individual interviews were conducted with GPs and resident GPs (n 20) in Swedish PHC and analysed using qualitative content analysis.

The overarching theme PEth testing in primary health care-a game changer and a challenge illustrated that PEth testing has improved the prerequisites for the GP-patient interaction while making it more complex. Four categories underpinned this theme: Comprehending the context, describing the challenges in the GP-patient interaction when hazardous alcohol use or AUD was suspected; Getting the pieces in place, illustrating the struggle of integrating PEth testing into clinical practice and how it diminished the role of alcohol history taking; The challenges and facilitators of the conversation, comprising both the difficulties in informing about PEth testing and the positive impact on the interaction, and Considerations based on the PEth test results, emphasising the consequences of elevated PEth test results and their influence on physicians' motivation to using PEth.

PEth is an important tool in the identification of hazardous alcohol use. Emerging ethical dilemmas regarding patient information on PEth testing and management of medical and medico-legal obligations when test results indicate high alcohol use need to be addressed in future guidelines for clinical management of PEth.

Phosphatidylethanol (PEth) consists of phospholipids synthesized in erythrocyte cell membranes in the presence of ethanol and serves as a sensitive and specific indicator of alcohol consumption. Further research on PEth formation, degradation, and stability in postmortem (PM) samples would support its routine application in forensic toxicology. A supercritical fluid chromatography-tandem mass spectrometry (SFC-MS-MS) method was developed and validated to quantify PEth 16:0/18:1 in blood. PEth 16:0/18:1 was extracted from blood (0.25 g) using an 8:2 (v/v) heptane:2-propanol mixture. Method validation results met American National Standards Institute/Academy Standards Board 036 guidelines. Recovery was >48%, and matrix effects were <20%. The linear range was 10-2500 ng/g, and lower limit of quantification was 10 ng/g. Bias was ±17.7%, and precision was <17.1% for all quality control levels. Carryover, endogenous, and exogenous interferences were negligible. Extracts were stable beyond 72 hours. In a proof-of-concept study reanalyzing 35 PM case samples, PEth concentrations ranged between 32.6 to 2476 ng/g. Short-term stability studies showed that fortified bovine blood (200 ng/g) preserved with 0.4% sodium fluoride (NaF) stored at room temperature had a 6.6% concentration drop after 48 hours, while blood stored at 4°C decreased by 13.5% over 14 days. Additionally, human PEth-positive blood preserved with 0.4% NaF showed a 6.7% decrease in in vivo PEth concentrations compared to a 17.5% decrease in heparin-preserved blood after 14 days at 4°C, supporting the use of 0.4% NaF in reducing PEth degradation over time. An in vitro model was also developed to simulate early PM PEth changes. Results found that PEth formation occurred in an ethanol concentration-dependent manner with minimal degradation, and considerations should be taken when interpreting PEth concentrations in cases with long PM interval, and if the decedent had a high blood alcohol concentration level and was left at elevated temperatures. This is the first SFC-MS-MS method successfully developed and validated for the analysis of PEth in PM samples.

Phosphatidylethanol (PEth) is a direct alcohol biomarker which is used in a variety of clinical and forensic contexts. Due to its limited stability in whole blood, PEth is routinely analysed using dried blood spots (DBS). There are many different commercially available sampling systems for DBS, but data on their comparability is scarce. The aim of this study was to develop and validate a LC-MS/MS method for the quantification of PEth 16:0/18:1 and PEth 16:0/18:2 for three different DBS sampling systems: Whatman™ filter paper (903 Protein Saver Cards), Mitra® and Capitainer®B Vanadate. The transferability of calibration curves and their applicability across these devices should be assessed. The results showed that DBS of all devices could be prepared and analysed using the same procedure. Validation was successful for the devices. The accuracy of QCs was best if quantified using the matching DBS sampling device calibration curve (bias < 15 %). Higher, but still acceptable, bias values were observed when using a Whatman™ calibration curve to quantify PEth in DBS of the Capitainer®B Vanadate system and vice versa. Mitra® QCs should only be quantified using device-specific calibration curves, as in all other cases, accuracy exceeded the limit of 15 %. We recommend analysing samples using device-specific calibrations at all times, as determined PEth concentrations might be crucial for patients and should not be subjected to any avoidable uncertainty.

Phosphatidylethanol (PEth) is an ethanol metabolite used as a specific biomarker for recent alcohol consumption. We aimed to determine the proportion of patients with or at risk for metabolic dysfunction-associated steatotic liver disease (MASLD) who had PEth levels indicative of harmful alcohol consumption, and to assess associations between PEth levels and the risk of major adverse liver outcomes (MALOs).

We conducted a cohort study involving persons tested for PEth in Stockholm, Sweden between 2012 and 2020 (N=46,406), including patients with various steatotic liver disease (SLD) subtypes and individuals without SLD. Cumulative incidences of MALOs were calculated for the different groups while accounting for competing risk. Cox regression was used to evaluate the association between baseline PEth levels and the incidence of MALOs.

Among 6,377 patients with presumed MASLD, 1,294 (20%) had baseline PEth levels between 0.05 and 0.30 μmol/L (35-210 ng/ml), indicating excessive alcohol intake (MetALD), while 854 patients (13%) had values >0.30 μmol/L, indicating alcohol-related liver disease (ALD). Patients with MASLD and PEth levels between 0.05-0.30 μmol/L had similar median FIB-4 scores and cirrhosis prevalence as those with MASLD and PEth levels <0.05 μmol/L. However, patients with PEth levels between 0.05-0.30 μmol/L had higher cumulative incidences of MALOs compared to those with PEth levels <0.05 μmol/L. Elevated PEth levels were significantly linked to higher rates of MALOs in patients without cirrhosis, even after adjustments for age, sex, SLD subtype, and FIB-4 score. Patients with ALD had the highest PEth levels and worst prognosis.

PEth is a valuable alcohol biomarker for distinguishing between SLD subtypes, especially ALD, and predicts adverse outcomes in people with and without SLD.

There is controversy regarding the various proposed steatotic liver disease (SLD) subtypes, the most recent definition suggesting that patients with an elevated alcohol consumption and MASLD should be classified as having MetALD. Here, we address this challenge by classifying patients with SLD by utilizing the biomarker phosphatidylethanol (PEth), a direct and reliable biomarker for recent alcohol consumption. Our analysis of this large cohort-comprising 46,406 patients-revealed that using the objective PEth biomarker may be a valuable tool for distinguishing between MASLD and MetALD, and that PEth is strongly associated with the risk of liver outcomes in individuals with and without known SLD. Integrating PEth testing into routine diagnostic evaluations could enhance knowledge on the underlying pathophysiology in SLD, reduce the potential for misclassification, and ultimately improve patient outcomes by enabling clinicians to offer appropriate therapies. Further research is needed to validate these findings in other populations and to explore the potential integration of PEth into broader clinical guidelines for managing SLD.

There is a growing interest for quantification of drugs in capillary blood. Phosphatidylethanol (PEth) is a biomarker for alcohol intake measured in whole blood, thus making it a candidate for capillary sampling. Our laboratory has been running a method for PEth quantification in venous blood since 2016, and we aimed to expand this method to also include capillary dried blood spot (DBS) samples. Two 10-µl volumetric absorptive microsampling (VAMS) devices, Capitainer®B Vanadate and Mitra®, were included in the method development and validated. Calibrators and quality controls were spiked during automatic sample extraction without the VAMS devices present, making it possible to extract and analyze both types of VAMS samples in the same setup. With the Mitra device, all pre-established validation criteria were fulfilled in the measuring range of 0.03 to 4.0 µM (21-2812 ng/mL), including method comparison with our venous blood method. Capitainer fulfilled all validation criteria, except for the accuracy of samples with PEth levels ≥ 0.5 µM (≥ 352 ng/mL) (deviation -17.1% to -20.5%). The correlation analysis between Capitainer and the venous blood results showed no constant bias, but an acceptable small proportional mean difference of -7.6%. Overall, the method validation results for both Capitainer and Mitra were considered acceptable. Both devices were found to be suitable for the analyses of PEth.

Alcohol use disorder is a major risk factor for alcohol-associated liver disease (ALD), characterized by reduced hepatic alcohol dehydrogenase (ADH) activity, increased body burden of alcohol, and its nonoxidative metabolism to fatty acid ethyl esters (FAEEs). However, the mechanism(s) underlying ALD remain unclear. This study investigated the metabolic basis and mechanism(s) of ALD in chronic ethanol (EtOH)-fed hepatic ADH1-deficient (ADH-) deer mice administered with a single dose of binge EtOH with/without FAEEs. Hepatic ADH- and ADH normal (ADH+) deer mice fed chronic EtOH daily for 3 mo, followed by a single dose of binge EtOH (3 g/kg·body wt) with/without FAEEs (100 mg/kg·body wt), 1 wk before euthanasia. Blood alcohol and acetaldehyde and liver injury markers in the plasma, hepatic FAEEs, lipids, and inflammatory markers were analyzed. Hepatic histology, ultrastructure, protein/mRNA expression of genes involved in alcohol metabolism and lipogenesis, cyclic adenosine monophosphate (cAMP), phosphodiesterase (PDE) activity, and AMP-activated protein kinase (AMPKα) signaling were assessed. Blood alcohol, hepatic lipids and FAEEs, inflammation, oxidative stress, and the expression of lipogenic proteins/genes were significantly increased in various chronic EtOH-fed groups of ADH- versus ADH+ deer mice. In addition, hepatic cAMP levels were reduced, whereas PDE activity and plasma transaminases were elevated. Binge EtOH with/without FAEEs did not significantly exacerbate the liver injury in chronic EtOH-fed ADH- as well as ADH+ deer mice. Overall, an increased body burden of EtOH and endogenously formed FAEEs due to hepatic ADH deficiency, along with dysregulated cAMP and AMPKα signaling, could be the determining factors for EtOH-induced liver injury leading to ALD.NEW & NOTEWORTHY Using hepatic alcohol dehydrogenase deficient (ADH-) deer mouse, which mimics the metabolic conditions observed in chronic alcoholics, we found significant hepatic injury along with degenerative changes in endoplasmic reticulum and mitochondria. Our findings suggest that an increased nonoxidative alcohol metabolism under hepatic alcohol dehydrogenase deficiency and associated hepatic lipid dysregulation and injury appear to be the key factors involved in the pathogenesis of alcohol-associated liver disease.

Alcohol consumption is widespread in most western countries such as Germany and a relevant risk factor for morbidity and mortality. Sensitive detection of alcohol consumption using suitable markers is therefore of central importance for clinical and forensic diagnostics. Direct alcohol markers are non-oxidative products of ethanol, which are produced in the body during the degradation of ethanol and provide high sensitivity and specificity. Phosphatidylethanol (PEth) is a promising marker for detecting alcohol consumption in the past days to weeks. The aim of this study was to determine the minimum amount of ethanol for a single alcohol consumption that leads to a detectable increase in blood PEth concentration. Therefore, 12 participants were recruited and, after four weeks of abstinence, drinking tests were carried out with target blood alcohol concentrations (BAC) of 0.6 g/kg and 0.75 g/kg. The PEth samples were obtained as dried-blood spots on the test day and the three following days and analyzed using LC-MS/MS. The result of the study were a detectable increase of PEth in the blood above limit of detection after both drinking events in all participants and an increase in PEth above the cutoff concentration for abstinence of 20 ng/mL in 9/12 (75%) and 7/12 (58%) participants, respectively, from a minimum BAC of 0.48 g/kg. These results make PEth appear promising as a marker for controlled moderate alcohol consumption.

Advancements with cost-effective, high-throughput omics technologies have had a transformative effect on both fundamental and translational research in the medical sciences. These advancements have facilitated a departure from the traditional view of human red blood cells (RBCs) as mere carriers of hemoglobin, devoid of significant biological complexity. Over the past decade, proteomic analyses have identified a growing number of different proteins present within RBCs, enabling systems biology analysis of their physiological functions. Here, we introduce RBC-GEM, one of the most comprehensive, curated genome-scale metabolic reconstructions of a specific human cell type to-date. It was developed through meta-analysis of proteomic data from 29 studies published over the past two decades resulting in an RBC proteome composed of more than 4,600 distinct proteins. Through workflow-guided manual curation, we have compiled the metabolic reactions carried out by this proteome to form a genome-scale metabolic model (GEM) of the RBC. RBC-GEM is hosted on a version-controlled GitHub repository, ensuring adherence to the standardized protocols for metabolic reconstruction quality control and data stewardship principles. RBC-GEM represents a metabolic network is a consisting of 820 genes encoding proteins acting on 1,685 unique metabolites through 2,723 biochemical reactions: a 740% size expansion over its predecessor. We demonstrated the utility of RBC-GEM by creating context-specific proteome-constrained models derived from proteomic data of stored RBCs for 616 blood donors, and classified reactions based on their simulated abundance dependence. This reconstruction as an up-to-date curated GEM can be used for contextualization of data and for the construction of a computational whole-cell models of the human RBC.

The use of phosphatidylethanol (PEth) as a biomarker for alcohol consumption is increasing likely due to its relatively long half-life in blood. Here, we present a pharmacokinetic model for three common homologs of PEth based on concentrations of each observed in a 5-day study of daily alcohol consumption. Adult participants were 11 females and 6 males with a median age of 32 years and median BMI of 24.3, all of whom drank on 1 or more days per week with at least 1 day per month of "heavy" drinking and also free from psychiatric disorders. All participants were abstinent for one week prior to beginning the study. The overall goals of this modeling effort are the use of PEth for assessment of alcohol consumption behavior and better understanding of the biological mechanisms underlying PEth pharmacokinetics. The modeling presented encompasses both the calibration of the pharmacokinetic model from daily individual PEth measurements and the prediction of model parameters in the study population with a regression model. The overall model was then evaluated by comparison of predicted PEth levels in blood with those measured in several groups of subjects in controlled drinking experiments. The results of this modeling effort indicate that the model can predict PEth concentrations in blood from alcohol consumption albeit with high variability both between individuals and within a single individual between drinking occasions. These results suggest the possible need to refine currently used cutoffs used in clinical and forensic contexts to predict alcohol consumption amounts. (249 words).

Alcohol withdrawal is frequently encountered in hospital settings, but the decision to treat the withdrawal with pharmacotherapy is often made without knowing if treatment is even necessary. Testing for phosphatidylethanol (PEth), results of which correlate well with recent alcohol consumption, may allow hospital teams to avoid treating alcohol withdrawal unnecessarily. To investigate PEth values of those suspected of alcohol withdrawal syndrome (AWS) among hospital inpatients, we conducted a retrospective study to determine the prevalence of hospitalized patients who received pharmacotherapy for withdrawal who also demonstrated PEth values suggesting minimal recent drinking.

This retrospective study examined the electronic health records of hospitalized adult patients suspected of recent heavy alcohol use for whom PEth tests were ordered. PEth tests were ordered at the time to inform treatment decisions such as the provision of naltrexone or referral to ongoing psychosocial treatments, but not to inform the decision to treat AWS given that results would only be available 3 or more days later. Hospital inpatients, excluding emergency room encounters, for whom PEth was ordered at 2 affiliated academic hospitals from January 2021 to April 2024 were included.

The included cohort was composed of 73 individual encounters. Among the 45 (62%) encounters where pharmacotherapy for AWS was administered, 14 (31%) had PEth values (≤200 ng/ml) inconsistent with heavy alcohol use.

Results suggest that a substantial proportion of hospital inpatients may have received pharmacotherapy for AWS despite PEth levels inconsistent with recent heavy alcohol use. If PEth results are available in real-time, results could help clinicians decide if pharmacotherapy is warranted. Further study of PEth testing and its role in the care of hospitalized patients is recommended.

Phosphatidylethanol (PEth) measurement in whole blood samples is established as a specific alcohol biomarker with clinical and forensic applications. Establishment of dried blood spots (DBSs) as a specimen for PEth determination offers several advantages and was the focus of this work. A liquid chromatography tandem mass spectrometry method using a 96-well format for sample preparation was developed and validated. PEth was extracted from DBSs by using isopropanol containing PEth-d5 as internal standard. The blood sampling used a commercial volumetric DBS device having a phosholipase D inhibitor incorporated to stop continuous PEth formation. The method quantified PEth in the range of 0.05-10 μmol/L, with a bias and imprecision of less than 15%. In a clinical study (n = 25) using fingerprick blood, the volumetric device offered more precise quantifications (CV 4.6%) compared with the Whatman 903 Protein Saver card device (CV 16.6%). In another clinical study (n = 48), the use of dried venous and capillary blood, and liquid venous blood was compared under real-life conditions with samples sent by postal service. The capillary and venous DBS samples gave identical results while the liquid blood gave slightly higher values. Calculation of elimination half-life (PEth 16:0/18:1) in 31 cases based on two consecutive samples with 2-9 days in between gave results (mean 6.2 days) that agree with literature but several cases with values over 10 days. In conclusion, this study demonstrates that volumetric DBS is a valid specimen for determination of PEth blood concentrations, offering several advantages.

Alcohol use disorder (AUD) is a massive burden for the individual, relatives and society. Despite this, the treatment gap is wide compared with other mental health disorders. Treatment options are sparse, with only three Food and Drug Administration (FDA)-approved pharmacotherapies. Glucagon-like peptide-1 (GLP-1) receptor agonists have shown promising effects in reducing alcohol consumption in preclinical experiments, and clinical trials are in high demand to investigate these potentially beneficial effects in patients diagnosed with AUD.

The effects of the once-weekly GLP-1 receptor agonist semaglutide will be investigated in a 26-week, randomised, placebo-controlled, double-blinded clinical trial. 108 patients diagnosed with AUD and comorbid obesity (body mass index (BMI)≥30 kg/m2)) will be randomised to treatment with either semaglutide or placebo in combination with cognitive behavioural therapy. A subgroup of the patients will have structural, functional and neurochemical brain imaging performed at baseline and after 26 weeks of treatment. The primary endpoint is the reduction in heavy drinking days, defined as days with excess consumption of 48/60 g of alcohol per day (women and men, respectively). Secondary endpoints include changes from baseline to week 26 in alcohol consumption, smoking status, quality of life, fibrosis-4 score, plasma concentration of phosphatidylethanol, brain gamma-aminobutyric acid (GABA) levels, alcohol cue reactivity, functional connectivity and white matter tract integrity.

Recruitment started in June 2023.

The study is approved by the Ethics Committee of the Capital Region of Denmark, the Danish Board of Health and the Danish Data Protection Agency. All patients will sign the written consent form before being included in the trial. Results will be disseminated through peer-reviewed publications and conference presentations. After the results are published, all de-identified data will be available in the Mendeley database.

NCT05895643.

Harmful alcohol use and alcohol use disorder (AUD) are common worldwide, and rates of alcohol-associated liver disease (ALD) are also increasing. AUD is a disease that is treatable and can be diagnosed and managed, and recovery from AUD through abstinence or reductions in drinking is possible. Management of AUD among individuals with ALD is increasingly being addressed via integrated medical and psychosocial treatment teams that can support reductions in drinking and prevent progression of liver disease. Early diagnosis of AUD and ALD can improve lives and reduce mortality.

The objective of this study was to identify the prevalence and correlates of phosphatidylethanol (PEth) levels suggestive of unhealthy alcohol use among women living with and without HIV who self-reported no or low-risk drinking. We analyzed data from a cross-sectional study among women enrolled in the San Francisco Bay Area site of the Women's Interagency HIV Study (WIHS). Between October 2017 and March 2018, PEth was tested from dried blood spots in 192 women enrolled in the San Francisco site of the WIHS. Using multivariable logistic regression, we identified the correlates of PEth levels suggestive of unhealthy alcohol use (>50 ng/ml) among the 168 women who reported no or low-risk drinking (<7 drinks per week) in the past six months, while controlling for age in years and race/ethnicity. Among the 168 women in the analysis sample, the median age was 55; 51% identified as Black/African American, 47% were living with HIV and 28% had PEth levels ≥50 ng/ml which are suggestive of unhealthy alcohol use. Factors independently associated with PEth levels ≥50 ng/ml in adjusted models were: identifying as Black/African American (adjusted odds ratio [aOR] = 8.34, 95% CI = 2.06-33.72), having an alanine transaminase to aspartate aminotransferase ratio > 1 (aOR = 3.10, 95% CI = 1.18-8.13), higher high-density lipoprotein levels (aOR = 1.31 per 10 mg/dL increase, 95% CI = 1.01-1.70), and consuming a greater number of drinks per week in the past six months (aOR = 1.40, 95% CI = 1.10-1.78). Nearly a third of women in this study had PEth levels suggestive of unhealthy alcohol use and potentially under-reported their use. To optimize alcohol related health care, there is a need to consider approaches to improve ascertainment of unhealthy alcohol use, especially among Black/African American women and those living with liver disease, so that interventions can be initiated.

Prenatal alcohol exposure (PAE) is one of the leading causes of preventable developmental disabilities. A lack of objective screening methods results in an under-recognition of the phenomenon. Phosphatidylethanol (PEth) is a specific ethanol biomarker that reveals alcohol intake up to several weeks after alcohol use. So far, PEth has mostly been a tool for detecting moderate and heavy drinking. With lower PEth cut-offs, revealing even minor prenatal alcohol consumption is possible. We aimed to find out if a sensitive method for PEth analysis would give additional information about PAE and to assess the cut-off value for a positive alcohol result in prenatal screening.

The study was an observational study of 3000 anonymous blood samples collected from the Helsinki University Hospital Diagnostic Center between June and September 2023. The Finnish Red Cross Blood Service received the samples originally for blood group typing and antibody screening as part of the prenatal blood screening program. We developed a sensitive PEth 16:0/18:1 analysis method using ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) equipment after liquid-liquid extraction of PEth from whole blood. The lower limit of quantification was 1 ng/mL.

PEth was ≥2 ng/mL in 5.2% of the cases, ≥8 ng/mL in 2.0%, and ≥20 ng/mL in 1.0%. The detection time of PEth can be several weeks, especially with low PEth concentrations and after heavy alcohol consumption. It remained unknown whether the positive PEth tests resulted from drinking deliberately during pregnancy or before pregnancy recognition.

We suggest adding PEth 16:0/18:1 to a routine prenatal blood screening program with a cut-off of 2 ng/mL-and in positive cases, clinical evaluation and retesting in 2-4 weeks. In clinical settings, information on gestational week and alcohol consumption before pregnancy is relevant and needs to be considered when interpreting low PEth concentrations.

Phosphatidylethanol (PEth) is a specific marker of alcohol intake, used both as a screening method for hazardous use and as an outcome measure in the treatment of alcohol use disorder (AUD). However, what cut-off values to apply for hazardous use in a treatment setting is still unclear. We aimed to investigate the correlation between PEth and self-reported drinking and identify the optimal cut-off for hazardous use, for patients with AUD and a stated goal of controlled drinking.

We used data from a randomized controlled trial of two different psychological treatments aiming for controlled drinking, conducted within specialized addiction care in Stockholm, Sweden. A total of 181 patients left samples that could be included in the current analysis. Outcomes were measured at five different time points over 2 years of follow-up. PEth 16:0/18:1 values were correlated with subjective reports of recent drinking based on the Timeline Follow-Back Method.

The correlation between PEth and self-reported alcohol intake increased significantly over time, with the weakest correlation found at baseline (Spearman's ρ = 0.42) and the strongest at the 104-week follow-up (ρ = 0.69). When used to indicate hazardous drinking according to Swedish guidelines (≥10 units per week), receiver operating characteristic analysis revealed PEth ≥ 0.22 μmol/l to be the optimal cut-off.

PEth is a useful outcome measure that can be used to validate subjective reports of current drinking. In a treatment setting aimed at controlled drinking, the accuracy of patients' self-report measures seems to improve over time. In this context, a PEth value of ≥0.22 μmol/l is a sensitive and specific indicator of hazardous drinking.

Alcohol use is associated with an increased incidence of negative health outcomes in burn patients due to biological mechanisms that include a dysregulated inflammatory response and increased intestinal permeability. This study used phosphatidylethanol (PEth) in blood, a direct biomarker of recent alcohol use, to investigate associations between a recent history of alcohol use and the fecal microbiota, short chain fatty acids, and inflammatory markers in the first week after a burn injury for nineteen participants. Burn patients were grouped according to PEth levels of low or high and differences in the overall fecal microbial community were observed between these cohorts. Two genera that contributed to the differences and had higher relative abundance in the low PEth burn patient group were Akkermansia, a mucin degrading bacteria that improves intestinal barrier function, and Bacteroides, a potentially anti-inflammatory bacteria. There was no statistically significant difference between levels of short chain fatty acids or intestinal permeability across the two groups. To our knowledge, this study represents the first report to evaluate the effects of burn injury and recent alcohol use on early post burn microbiota dysbiosis, inflammatory response, and levels of short chain fatty acids. Future studies in this field are warranted to better understand the factors associated with negative health outcomes and develop interventional trials.

This article traces the historical development of various biomarkers of acute and/or chronic alcohol consumption. Much of the research in this domain of clinical and laboratory medicine arose from clinics and laboratories in Sweden, as exemplified by carbohydrate deficient transferrin (CDT) and phosphatidylethanol (PEth). Extensive studies of other alcohol biomarkers, such as ethyl glucuronide (EtG), ethyl sulfate (EtS), and 5-hydroxytryptophol (5-HTOL), also derive from Sweden. The most obvious test of recent drinking is identification of ethanol in a sample of the person's blood, breath, or urine. However, because of continuous metabolism in the liver, ethanol is eliminated from the blood at a rate of 0.15 g/L/h (range 0.1-0.3 g/L/h), so obtaining positive results is not always possible. The widow of detection is increased by analysis of ethanol's non-oxidative metabolites (EtG and EtS), which are more slowly eliminated from the bloodstream. Likewise, an elevated ratio of serotonin metabolites in urine (5-HTOL/5-HIAA) can help to disclose recent drinking after ethanol is no longer measurable in body fluids. A highly specific biomarker of hazardous drinking is CDT, a serum glycoprotein (transferrin), with a deficiency in its N-linked glycosylation. Another widely acclaimed biomarker is PEth, an abnormal phospholipid synthesized in cell membranes when people drink excessively, having a long elimination half-life (median ~6 days) during abstinence. Research on the subject of alcohol biomarkers has increased appreciably and is now an important area of drug testing and analysis.

The use of direct alcohol biomarkers (ethylglucuronide and phosphatidylethanol) has recently been implemented in a clinical setting. Due to their low alcohol detection threshold, high sensitivity, and specificity, these tools are very useful in the pre- and post-liver transplantation setting, where the history and physical signs are not always reliable. However, the interpretation of the results can sometimes be misleading and must be integrated into a global clinical evaluation and, more importantly, in the clinical context of each patient. We present here a case report illustrating a false-positive hair ethylglucuronide caused by the application of a capillary gel in an abstinent patient after liver transplantation. This reminds us that even the most accurate laboratory tests must be interpreted with caution.

When identifying, preventing and treating alcohol use disorder, a correct estimation of alcohol intake is essential. An objective marker is preferred as self-reported alcohol intake suffers from bias, and the use of alcohol biomarkers is increasing globally. An easy-to-use blood biomarker to correctly assess alcohol consumption is an invaluable asset in alcohol treatment strategies, as well as in alcohol research studies. The specific, cumulative, biomarker phosphatidylethanol, mirroring the past two weeks of consumption, has shown superiority over traditional biomarkers and is an attractive choice of proxy for alcohol intake.

Alcohol-related liver disease is the most common indication for liver transplantation. It is essential for providers in transplantation to be informed of the state of the science in evaluation of alcohol use disorder (AUD). This review examines the broad range of approaches to the evaluation of AUD ranging from traditional interview approaches to recent literature on artificial intelligence models. The empirical support for methods of evaluation is examined. The authors discuss the use of each method in the context of patients seeking a liver transplant for alcohol-related liver disease. This review emphasizes the importance of using objective assessments so that transplant centers make evidence-based decisions and reduce cognitive bias. The review concludes with a proposed assessment battery for evaluation and bridges to future directions in the field of AUD assessment in liver transplantation.

Alcohol-associated liver disease (ALD) has emerged as the leading indication for liver transplantation (LT) worldwide, with 40% of LTs in the United States performed for ALD in 2019. The ALD-related health care burden accelerated during the COVID-19 pandemic, especially in young individuals. Alcohol use disorder (AUD), which focuses on the negative effects of alcohol on psychosocial, physical, and mental health, is present in the majority of patients with ALD, with moderate to severe AUD in 75%-80%. During the last decade, early liver transplantation (eLT) has emerged as a lifesaving treatment for selected patients with alcohol-associated hepatitis; these patients may have a higher risk of using alcohol after LT. The risk of alcohol use recurrence may be reduced during the pretransplant or post-transplant period with AUD treatment using behavioral and/or pharmacological therapies and with regular monitoring for alcohol use (self-reported and complemented with biomarkers like phosphatidylethanol). However, AUD treatment in patients with ALD is challenging due to patient, clinician, and system barriers. An integrated model to provide AUD and ALD care by hepatologists and addiction experts in a colocated clinic starting from LT evaluation and selection to monitoring listed candidates and then to following up on recipients of LT should be promoted. However, the integration of addiction and hepatology teams in an LT program in the real world is often present only during evaluation and candidate selection for LT. Data are emerging to show that a multidisciplinary integrated AUD treatment within an LT program reduces recurrent alcohol use after LT. If we want to continue using early liver transplantation for patients with severe alcohol-associated hepatitis, LT programs should focus on building integrated multidisciplinary care teams for the integrated treatment of both AUD and ALD.

Delayed CD4 recovery after initiating antiretroviral therapy (ART) is a novel potential mechanism by which alcohol consumption leads to increased morbidity and mortality in people with HIV. We hypothesized that alcohol consumption at ART initiation is associated with slower CD4 recovery.

We retrospectively analyzed 2 pooled longitudinal alcohol/HIV cohorts (2014-2019) in St. Petersburg, Russia. Eligible participants initiated the first ART during parent studies; had alcohol consumption assessed by the blood biomarker, phosphatidylethanol (PEth), at the last research visit before ART initiation; and had ≥1 CD4 count measurement before and after initiating ART. Participants were stratified by low, moderate, and high PEth (<8, 8-80, and >80 ng/mL, respectively). We used random-effects piecewise linear regression models to estimate CD4 recovery, defined as CD4 count change per 30 days after ART initiation, by the alcohol group.

Of 60 eligible participants, median age was 34 years and 28% were female. The median pre-ART PEth in the low, moderate, and high PEth groups were <8, 23, and 232 ng/mL, respectively. After starting ART, the CD4 count increased by 13.60 cells/mm3/mo (95% CI: 0.33 to 26.87) with low PEth, 0.93 cells/mm3/mo (95% CI: -6.18 to 8.04) with moderate PEth, and 2.33 cells/mm3/mo (95% CI: -3.44 to 8.09) with high PEth.

Among Russians with HIV, we observed faster CD4 recovery after ART initiation in those with low alcohol consumption compared with those with moderate and high alcohol consumption, as assessed by PEth. This analysis provides further evidence for the possible value of alcohol reduction interventions for people with HIV who are initiating ART.

Phosphatidylethanol (PEth) are membrane molecules formed from phosphatidylcholine and ethanol through transphosphatidylation catalyzed by phospholipase D. Measurement of the main PEth form 16:0/18:1 is used as a specific and sensitive alcohol biomarker, since its formation requires ethanol, it accumulates in the blood upon repeated ethanol exposure, and it is only slowly eliminated during abstinence. PEth formation correlates with alcohol intake at the population level, albeit with considerable inter-individual variation as for the half-life during withdrawal. Over the past decade, the use of PEth has increased significantly and the applications have broadened. In Sweden, routine decision limits and the interpretation of test results for PEth were harmonized in 2013, using < 0.05 µmol/L (∼35 µg/L) as the recommended lower reporting limit and values > 0.30 µmol/L (∼210 µg/L) to indicate regular high alcohol intake. Routine test results show a large variation with about half being < 0.05 µmol/L and some even exceeding 10 µmol/L. In 2013, an external quality assessment (EQA) scheme for PEth 16:0/18:1 measurement in whole blood was also started (Equalis, Uppsala, Sweden), presently involving 56 laboratories from 13 countries. The agreement of PEth results between the laboratories has gradually improved to a CV < 15%. The current clinical and scientific information suggests that PEth values below the lower reporting limit (typically ∼0.03-0.05 µmol/L, or ∼20-35 µg/L) indicates sobriety or only low or occasional alcohol consumption, while regular high alcohol intake at levels corresponding to harmful drinking is required in most cases to reach PEth values > 0.30 µmol/L.

Phosphatidylethanol (PEth) is a group of phospholipids formed exclusively in the presence of ethanol on the erythrocyte membrane, making it a direct biomarker for long-term ethanol consumption for which a clinical reference interval has been established. Here, we describe an assay for quantitation for two most abundant PEth homologues, PEth 16:0/18:1 and PEth 16:0/18:2, from human whole blood, and present challenges overcome throughout the development process. Since PEth is localized within erythrocyte membranes, a reliable sample preparation technique is an important aspect of PEth analysis. Therefore, various erythrocyte lysing agents for recovery of exogenously spiked standards and controls were evaluated to identify one that performed comparably to the recovery of endogenous analytes found in authentic samples. A supported liquid extraction (SLE) technique was employed for sample cleanup and enrichment which together with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis enabled automated sample preparation, appropriate chromatographic resolution, and minimal system carryover. This resulted in a laboratory developed test with an analytical measurement range (AMR) of 10-1000 ng/mL (slope = 0.9902-1.0138, R2 = 0.9958-0.9972), that was precise (intra-day precision: 3.4-4.1%; inter-day precision: 4.4-8.2% over the AMR), accurate when compared with an available external laboratory test (slope = 0.9943-1.0206, R2 = 0.9635-0.9678, no lower decision point interpretation changes), with effective analyte recovery (77.2-83.5%), and established stability characteristics, while chromatographically separating the analytes to ensure no additive effects due to the isotopic distribution of the opposing analyte.

Detection of alcohol (ETOH) use with biomarkers provides an opportunity to intervene and treat patients with alcohol use disorder before and after liver transplant (LT). We describe our center's experience using urine ethyl glucuronide (EtG) and serum phosphatidylethanol (PEth) in alcohol screening protocols.

Single-center, retrospective review of patients presenting for LT evaluation, patients waitlisted for LT for alcohol-associated liver disease (ALD), and patients who received a LT for ALD over a 12-month period, from October 1, 2019 through September 30, 2020. Patients were followed from waitlisting to LT, or for up to 12 months post-LT. We monitored protocol adherence to screening for ETOH use- defined as completion of all possible tests over the follow-up period- at the initial LT visit, while on the LT waitlist and after LT.

During the study period, 227 patients were evaluated for LT (median age 57 years, 58% male, 78% white, 54.2% ALD). Thirty-one patients with ALD were placed on the waitlist, and 38 patients underwent LT for ALD during this time period. Protocolized adherence to screening for alcohol use was higher for PEth for all LT evaluation patients (191 [84.1%] vs. 146 [67%] eligible patients, p < .001), in patients with ALD waitlisted for LT (22 [71%] vs. 14 (48%] eligible patients, p = .04) and after LT for ALD, 20 (33 [86.8%] vs. 20 [52.6%] eligible patients, p < .01). Few patients with a positive test in any group completed chemical dependency treatment.

When screening for ETOH use in pre- and post-LT patients, protocol adherence is higher using PEth compared to EtG. While protocolized biomarker screening can detect recurrent ETOH use in this population, engagement of patients into chemical dependency treatment remains challenging.

The free-access (FA) intravenous alcohol self-administration (IV-ASA) paradigm is an experimental approach that can identify modulators of alcohol consumption in humans. Moreover, the outcome measures of IV-ASA paradigms are associated with self-reported alcohol intake using the timeline follow-back method (TLFB). To evaluate how FA IV-ASA reflects drinking in real life, we examined the relationship between an objective marker of recent alcohol intake, phosphatidylethanol in blood (B-PEth), and TLFB and measures obtained during IV-ASA in individuals with alcohol use disorder (AUD) and social drinkers (SD). We also explored the associations between these measures and gut-brain peptides involved in AUD pathophysiology.

Thirty-eight participants completed a laboratory session in which they self-administered alcohol intravenously. The safety limit was 200 mg%, and main outcomes were mean and peak breath alcohol concentrations (BrAC). Blood samples were drawn prior to IV-ASA and subjective alcohol effects were rated during the experiment.

The study sample comprised 24 SD and 14 participants with DSM-5 mild AUD. Although BrACs were not associated with B-PEth or TLFB in the full sample or AUD subgroup, there was an association with TLFB in SD. In both subgroups, BrACs were associated with alcohol craving but with differential timing. Total ghrelin levels were higher in AUD participants than in SD.

No associations between B-PEth levels and achieved BrACs were observed in the mild AUD group, the SD group, or the full sample. The ability for FA IV-ASA to reflect recent drinking was confirmed only for TLFB in SD, whereas there were no associations within the smaller subsample of participants with mild AUD or in the full sample. Further studies that include a larger AUD sample are warranted. The association of BrACs with craving for alcohol suggests that the IV-ASA method may be useful for assessing interventions that target craving. This could be explored by using the FA IV-ASA model to evaluate the effects on craving of approved pharmacotherapies for AUD.

The evaluation of suspected hypercortisolism is one of the most challenging problems in medicine. The signs and symptoms described by Dr Harvey Cushing are common and often create diagnostic confusion to even experienced endocrinologists. Cushing syndrome is classically defined as neoplastic hypercortisolism resulting from an ACTH-secreting tumor or from autonomous secretion of excess cortisol associated with benign or malignant adrenal neoplasia. The increasing recognition of the negative cardiometabolic effects of mild cortisol excess without overt physical signs of Cushing syndrome has led to more screening for endogenous hypercortisolism in patients with adrenal nodular disease, osteoporosis, and the metabolic syndrome. However, sustained or intermittent activation of the dynamic hypothalamic-pituitary-adrenal axis caused by chemical (alcohol), inflammatory (chronic kidney disease), psychologic (major depression), and physical (starvation/chronic intense exercise) stimuli can result in clinical and/or biochemical features indistinguishable from neoplastic hypercortisolism. Nonneoplastic hypercortisolism (formerly known as pseudo-Cushing syndrome) has been recognized for more than 50 years and often causes diagnostic uncertainty. This expert consultation describes two patients with features of Cushing syndrome who were referred for inferior petrosal sinus sampling for the differential diagnosis of ACTH-dependent hypercortisolism. Both patients were discovered to have nonneoplastic hypercortisolism: one from a covert alcohol use disorder and the other to chronic kidney disease. This consultation emphasizes the value of a good history and physical examination, appropriate laboratory testing, and the desmopressin acetate stimulation test to aid in distinguishing neoplastic from nonneoplastic hypercortisolism.

Alcohol-induced hypercortisolism (AIH) is underrecognized and may masquerade as neoplastic hypercortisolism [Cushing syndrome (CS)] obscuring its diagnosis.

In order to characterize AIH, we performed a chart review of eight patients (4 males and 4 females; 2014-2022) referred for evaluation and treatment of neoplastic hypercortisolism - six for inferior petrosal sinus sampling, one due to persistent CS after unilateral adrenalectomy, and one for pituitary surgery for Cushing disease (CD). Five underwent dDAVP stimulation testing.

All eight patients had clinical features of hypercortisolism and plasma ACTH levels within or above the reference interval confirming hypothalamic-pituitary mediation. All had abnormal low-dose dexamethasone suppression test and increased late-night salivary cortisol. Only one had increased urine cortisol excretion. In contrast to CD, the 5 patients tested had blunted or absent ACTH and cortisol responses to desmopressin. Two had adrenal nodules and one had abnormal pituitary imaging. Most patients underreported their alcohol consumption and one denied alcohol use. Elevated blood phosphatidyl ethanol (PEth) was required in one patient to confirm excessive alcohol use. All patients had elevations of liver function tests (LFTs) with AST>ALT.

AIH is an under-appreciated, reversible cause of non-neoplastic hypercortisolism that is indistinguishable from neoplastic CS. Incidental pituitary and adrenal imaging abnormalities as well as under-reporting of alcohol consumption further confound the diagnosis. Measurement of PEth helps to confirm an alcohol use disorder. Elevations of LFTs (AST>ALT) and subnormal ACTH and cortisol responses to dDAVP help to distinguish AIH from neoplastic hypercortisolism.

The medical disorders of alcoholism rank among the leading public health problems worldwide and the need for predictive and prognostic risk markers for assessing alcohol use disorders (AUD) has been widely acknowledged. Early-phase detection of problem drinking and associated tissue toxicity are important prerequisites for timely initiations of appropriate treatments and improving patient's committing to the objective of reducing drinking. Recent advances in clinical chemistry have provided novel approaches for a specific detection of heavy drinking through assays of unique ethanol metabolites, phosphatidylethanol (PEth) or ethyl glucuronide (EtG). Carbohydrate-deficient transferrin (CDT) measurements can be used to indicate severe alcohol problems. Hazardous drinking frequently manifests as heavy episodic drinking or in combinations with other unfavorable lifestyle factors, such as smoking, physical inactivity, poor diet or adiposity, which aggravate the metabolic consequences of alcohol intake in a supra-additive manner. Such interactions are also reflected in multiple disease outcomes and distinct abnormalities in biomarkers of liver function, inflammation and oxidative stress. Use of predictive biomarkers either alone or as part of specifically designed biological algorithms helps to predict both hepatic and extrahepatic morbidity in individuals with such risk factors. Novel approaches for assessing progression of fibrosis, a major determinant of prognosis in AUD, have also been made available. Predictive algorithms based on the combined use of biomarkers and clinical observations may prove to have a major impact on clinical decisions to detect AUD in early pre-symptomatic stages, stratify patients according to their substantially different disease risks and predict individual responses to treatment.

Chronic and heavy alcohol consumption is commonly observed in alcohol use disorder (AUD). AUD often leads to alcohol-associated organ injury, including alcohol-associated liver disease (ALD). Approximately 10-20% of patients with AUD progress to ALD. Progression of ALD from the development phase to more advanced states involve the interplay of several pathways, including nutritional alterations. Multiple pathologic processes have been identified in the progression and severity of ALD. However, there are major gaps in the characterization and understanding of the clinical presentation of early-stage ALD as assessed by clinical markers and laboratory measures. Several Institutions and Universities, including the University of Louisville, in collaboration with the National Institutes of Health, have published a series of manuscripts describing early-stage ALD over the past decade. Here, we comprehensively describe early-stage ALD using the liver injury and drinking history markers, and the laboratory biomarkers (with a focus on nutrition status) that are uniquely involved in the development and progression of early-stage ALD.

Phosphatidylethanol 16:0/18:1 (PEth), found in whole blood, is a biomarker for alcohol consumption with high sensitivity, specificity, and a long detection window. The TASSO-M20 device is used to self-collect capillary blood from the upper arm and has advantages over finger stick methods. The purpose of this study was to (1) validate PEth measurement using the TASSO-M20 device, (2) describe the TASSO-M20 for blood self-collection during a virtual intervention, and (3) characterize PEth, urinary ethyl glucuronide (uEtG) and self-reported alcohol in a single participant over time.

PEth levels in blood samples dried on TASSO-M20 plugs were compared to those in (1) liquid whole blood (N = 14) and (2) dried blood spot cards (DBS; N = 23). Additionally, the self-reported drinking, positive or negative uEtG results (dip card cutoff ≥300 ng/mL), and observed self-collection of blood with TASSO-M20 devices for PEth levels were obtained over time during virtual interviews of a single contingency management participant. High-performance liquid chromatography with tandem mass spectrometry detection was used to measure PEth levels for both preparations.

PEth concentrations from dried blood on TASSO-M20 plugs and liquid whole blood were correlated (0 to 1700 ng/mL; N = 14; r2  = 0.988; slope = 0.951) and in a subgroup of samples with lower concentrations (N = 7; 0 to 200 ng/mL; r2  = 0.944, slope = 0.816). PEth concentrations from dried blood on TASSO-M20 plugs and DBS were correlated (0 to 2200 ng/mL; N = 23; r2  = 0.927; slope = 0.667) and in a subgroup of samples with lower concentrations (N = 16; 0 to 180 ng/mL; r2  = 0.978, slope = 0.749). Results of the contingency management participant indicate that changes in PEth levels (TASSO-M20) and uEtG concentrations were consistent with each other and with changes in self-reported alcohol use.

Our data support the utility, accuracy, and feasibility of using the TASSO-M20 device for blood self-collection during a virtual study. The TASSO-M20 device had multiple advantages over the typical finger stick method, including consistent blood collection, participant acceptability, and less discomfort as indicated by acceptability interviews.

Phosphatidylethanol (PEth) forms in erythrocyte membranes after alcohol consumption, offering a persisting biomarker, that is measurable in whole blood, washed erythrocytes and dried blood spots. For a predominantly erythrocyte-restricted analyte, erythrocyte concentrations seem to have most validity in patients who are anemic through alcoholism or other pathologies, despite preparation increasing assay complexity. Differences in specimen preparation alter PEth concentrations for the same patient, meaning that criteria for interpreting PEth results should relate to specimen type, presenting a barrier to achieving harmonization. We therefore tested whether erythrocyte PEth might be validly calculated by hematocrit correction of a whole blood PEth measurement. PEth testing primarily serves to distinguish drinkers from non-drinkers. In choosing between specimen types, it is important to compare their utility in separating those two groups. We therefore processed 281 blood samples from 17 non-drinkers and 61 drinkers, to prepare matched whole blood and washed erythrocyte specimens. These were assayed by liquid chromatography-tandem mass spectrometry and compared in identifying alcohol consumption. The erythrocyte PEth concentration in the whole blood specimens was also calculated by correcting whole blood concentration by the specimen's hematocrit, as an alternative to prepare washed erythrocytes. The hematocrit-corrected erythrocyte concentrations were included in these comparisons. Predictably, this work found that sensitivity was consistently better at the lower cut-off of 8 µg/L than at 20 µg/L. Sensitivities were also higher for washed erythrocytes than whole blood, explained by the lower erythrocyte mass in the same volume of whole blood. Hematocrit-corrected whole blood PEth concentrations correlated with erythrocyte concentrations, except for the four highest values, which did not influence comparative sensitivity. Specificity was 100% for washed erythrocytes, whole blood and hematocrit-corrected whole blood at either cut-off because non-drinkers had undetectable PEth. We conclude that hematocrit correction of whole blood PEth concentrations theoretically provides an alternative to the preparation of washed erythrocytes.

Phosphatidylethanol (PEth) has become a widespread marker offering an up to 4-week retrospective window to detect alcohol use. Due to the pandemic of coronavirus disease 2019, ethanol-based hand sanitizers are frequently used. The aim of this study was to develop and validate a method for the determination of up to seven different homologues of PEth from dried blood spots (DBSs) after use of an ethanol-based hand sanitizer. The objectives of its preliminary application were to prove whether a threshold of 20 ng/mL for PEth 16:0/18:1 is reached and whether other homologues are formed as well as if positive findings of urinary ethyl glucuronide (UEtG) can be observed with respect to assess monitoring of abstinence control programs. Ten volunteers (8 occasional and 2 regular drinkers) were recruited to excessively use an ethanol-based hand sanitizer on 5 successive days. DBSs and urine samples were collected daily. PEth and UEtG were determined by liquid chromatography--tandem mass spectrometry. In total, two volunteers with initial PEth 16:0/18:1 concentrations of 19.3 and 14.6 ng/mL exceeded the threshold of 20 ng/mL six times. Subjects drinking daily or almost daily had starting PEth 16:0/18:1 concentrations of 242 and 354 ng/mL, showing a decline of PEth concentrations in six out of the seven homologues over 5 days. In teetotalers, formation of PEth species could not be observed. Thus, not satisfying requirements in an alcohol monitoring program with initial PEth-negative blood cannot be explained by a frequent use of ethanol-based hand sanitizer only. In cases of regular alcohol consumption, PEth homologues are not likely to be further influenced. However, results indicated that individuals with a PEth concentration close to 20 ng/mL are at risk of exceeding the threshold by using ethanol-based hand sanitizer.

We demonstrate proof-of-concept for point-of-care assessment of long-term alcohol consumption by measuring phosphatidylethanol in blood/dried blood spots with nano-electrospray ionization and MS/MS using a miniature mass spectrometer. 'Abstinence', 'moderate', and 'chronic' consumption could be distinguished rapidly for both sample types, and quantitative performance was obtained with blood (LoQ-100 ng mL^-1).

Alcohol intake is an important risk factor for cardiovascular disease， liver disease， and diabetes. The accurate and objective evaluation of alcohol intake is important for disease prevention and intervention， as well as alcohol intake monitoring. Phosphatidylethanol （PEth） is a potential clinical biomarker of alcohol consumption. Monitoring PEth levels can provide an objective and quantitative basis for alcohol intake studies. Unlike other current alcohol biomarkers， PEth can only be produced in the presence of alcohol. Therefore， PEth is highly specific for alcohol intake and not affected by confounding factors， such as age， gender， hypertension， kidney disease， liver disease， and other comorbidities. Because of its long half-life and high specificity for alcohol intake， PEth may be used as a tool for monitoring drinking behavior in the clinical， transportation， and other fields. Given rapid developments in mass spectrometry technology over the past decade， liquid chromatography-tandem mass spectrometry （LC-MS/MS） has become the preferred method for PEth detection. However， most current LC-MS/MS methods focus on the determination of one or several PEth homologs， and the number of PEth homologs that can be determined simultaneously is relatively limited. Moreover， the detection capacity of the available methods remains insufficient， and their analytical sensitivity for some PEth homologs must be further improved. In this study， a novel LC-MS/MS method based on an intelligent scheduled time-zone negative multiple reaction monitoring acquisition technology （Scheduled-MRM） was developed. The technology monitors two ion channels in each PEth to ensure reliable results and can quantify 18 PEth homologs in human whole blood simultaneously. Methanol-methyl tert-butyl ether-water was used as the extraction system. An XBridge C18 column （100 mm×2.1 mm， 3.5 μm） was selected for gradient elution with 2.5 mmol/L ammonium acetate isopropanol solution and 2.5 mmol/L ammonium acetate aqueous solution-acetonitrile （50∶50， v/v） as the mobile phases. Negative electronic spray ionization in scheduled-MRM mode was applied for MS/MS detection. The method was validated to have a linear range of 10-2500 ng/mL with correlation coefficients greater than 0.9999. The limits of detection and quantification were 0.7-2.8 and 2.2-9.4 ng/mL， respectively， and the spiked recoveries ranged from 91.0% to 102.2%. The method was confirmed to be simple， rapid， and precise， and subsequently validated for the measurement of 18 PEth homologs in human blood. Scheduled-MRM can assign a suitable scan time to each ion channel and effectively reduce the number of concurrent ion pairs monitored per unit time. This technology overcomes the problem of insufficient dwell time caused by an excessive number of ion channels， thereby avoiding the redundant monitoring of non-retention times. Scheduled-MRM significantly improved the detection sensitivity， data acquisition quality， and signal response of the proposed method. Whole blood samples from 359 volunteers with regular drinking habits were analyzed using this method. The total PEth concentrations ranged from 51.13 ng/mL to 2.89 μg/mL， with a mean of 363.16 ng/mL. PEth 16∶0/18∶1 and 16∶0/18∶2 were the two most abundant homologs， with mean concentrations of 74.21 and 48.75 ng/mL， accounting for approximately 20.43% and 13.42%， respectively， of the total PEth. Spearman correlation analyses showed that the PEth homologs correlated well with each other， γ-glutamyltransferase， a clinically available biological marker of alcohol， and other clinical biochemical parameters related to liver and kidney function. Overall， the method was demonstrated to be sensitive， precise， and accurate； thus， it may be an effective tool for monitoring alcohol intake in the clinical and other fields.

The risk of alcohol use disorder increases after bariatric surgery. Preoperative alcohol use is a risk factor, and this is evaluated during the routine preoperative psychosocial evaluation. However, it is not clear whether patients accurately report their alcohol use.

To determine whether an objective measure of alcohol use, phosphatidylethanol (PEth) testing, offers utility beyond self-reported alcohol use during the preoperative evaluation for bariatric surgery.

Single healthcare system.

PEth testing was included as part of the routine laboratory work for 139 patients undergoing evaluation for bariatric surgery. PEth testing results were compared with self-reported alcohol use and scores on the Alcohol Use Disorders Identification Test-Concise (AUDIT-C) questionnaire obtained during the preoperative psychosocial evaluation. PEth testing results were categorized into abstinent, light use, moderate use, or heavy use. There were 85 patients who completed both PEth testing and a preoperative psychosocial evaluation.

There were 25 participants (29.4%) who had a positive PEth test; about half had moderate or heavy use values (15.3% of the total sample). The majority of participants with a positive PEth test (82.6%) denied recent alcohol use. Of those with PEth values indicating moderate or heavy use, 61.5% did not have an elevated AUDIT-C score.

Patients appeared to underreport their alcohol use during the preoperative psychosocial evaluation. There appears to be utility for routine PEth testing as part of the evaluation process to identify those with risky drinking patterns. Patients with preoperative risky drinking could be educated about their risk and/or referred to programs to mitigate the development of preoperative alcohol misuse.

In the literature on alcohol use biomarkers, there has been debate as to what a valid and/or utilitarian cut off level should be for various research applications. In this manuscript, we assessed the sensitivity and specificity of multiple cutoff values for phosphatidylethanol (PEth) from bloodspots relative to self-report, the Alcohol Use Disorder Identification Test (AUDIT) scores, and another alcohol use biomarker ethyl glucuronide (EtG) from fingernails in a sample of 222 pregnant women in the Western Cape Province of South Africa. Receiver operating characteristic (ROC) curves were used to assess the area under the curve (AUC) and assess PEth cutoff values of ≥2, ≥4, ≥8, ≥14, and ≥20 nanograms per milliliter (ng/ml). The highest AUC value was attained when PEth was compared to an AUDIT score of 1 or more. Depending on the cutoff used to determine alcohol consumption, PEth identified 47%-70% of the individuals as alcohol-consuming while 62.6%-75.2% were identified by self-reported measures, and 35.6% were identified by EtG. In this sample, sensitivity and accuracy were highest at less stringent PEth cutoffs when compared to self-report, AUDIT score of 1 or more, 5 or more, 8 or more, and EtG ≥ 8 picograms per milligram (pg/mg). For research purposes, less stringent cutoffs, such as PEth ≥ 8 ng/ml, may be considered a valid, positive cutoff for identifying women who consume alcohol during pregnancy in this population. A cutoff of PEth ≥ 20 ng/ml may miss individuals who reported consuming alcohol (false negatives).

People living with HIV (PLWH) represent a vulnerable population to adverse musculoskeletal outcomes due to HIV infection, antiretroviral therapy (ART), and at-risk alcohol use. Developing measures to prevent skeletal degeneration in this group requires a grasp of the relationship between alcohol use and low bone mass in both the PLWH population and its constituents as defined by sex, age, and race. We examined the association of alcohol use with serum biochemical markers of bone health in a diverse cohort of PLWH enrolled in the New Orleans Alcohol Use in HIV (NOAH) study. To explore the effects of alcohol on bone in the context of HIV and ART and the role of estrogen, we conducted a parallel, translational study using simian immunodeficiency virus (SIV)+/ART+ female rhesus macaques divided into four groups: vehicle (Veh)/Sham; chronic binge alcohol (CBA)/Sham; Veh/ovariectomy (OVX); and CBA/OVX. Clinical data showed that both osteocalcin (Ocn) and procollagen type I N-propeptide (PINP) levels were inversely associated with multiple measures of alcohol consumption. Age (>50 years) significantly increased susceptibility to alcohol-associated suppression of bone formation in both female and male PLWH, with postmenopausal status appearing as an additional risk factor in females. Serum sclerostin (Scl) levels correlated positively with measures of alcohol use and negatively with Ocn. Micro-CT analysis of the macaque tibias revealed that although both CBA and OVX independently decreased trabecular number and bone mineral density, only OVX decreased trabecular bone volume fraction and impacted cortical geometry. The clinical data implicate circulating Scl in the pathogenesis of alcohol-induced osteopenia and suggest that bone morphology can be significantly altered in the absence of net change in osteoblast function as measured by serum markers. Inclusion of sophisticated tools to evaluate skeletal strength in clinical populations will be essential to understand the impact of alcohol-induced changes in bone microarchitecture. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Exploration of the real-time relationship between substance use and delay discounting may reveal potential mechanisms driving high-risk behaviors. We conducted an ecological momentary assessment (EMA) study to investigate the effects of substance use on delay discounting in a sample of people who use stimulants (HIV+: 30; HIV-: 34). Participants completed multiple EMAs throughout the day for 28 days. The EMAs collected data on delay discounting and substance use (time since last substance use and level of intoxication). Delay discounting was assessed using a brief Monetary Choice Questionnaire (MCQ). Analyses were conducted using linear mixed effects modeling. Most participants (99.1%) used cocaine as their primary stimulant. Among participants without HIV, MCQ score remained relatively stable during the first 2 hr after stimulant use, followed by an increase during 2-6 hr (p < .05), before decreasing again. For alcohol and marijuana, the MCQ score was stable during the first 4 hr after use, with a sharp increase at 4-6 hr (p < .05), before decreasing again. Among participants with HIV, there were no changes in MCQ score as a function of time since recent substance use. These findings provide evidence of a plausible connection between delay discounting and acute withdrawal that may have relevance for risky behaviors. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Alcohol biomarkers can monitor both recent and long-term drinking and provide information about drinking habits as a complement to self-reporting. Ethyl glucuronide (EtG) and phosphatidylethanol (PEth) are the most sensitive available biomarkers for this purpose. The present study aimed to collect data on both PEth and EtG in the same blood sample, in addition to ethanol, in order to evaluate the combined use of these biomarkers. Venous EDTA blood samples (n = 1149) sent to the laboratory as part of a clinical routine service for measuring PEth were investigated. PEth and EtG concentrations were analyzed using liquid chromatography-mass spectrometry methods and ethanol with an enzymatic method. Of the 1149 samples, 95 were positive for ethanol (range 0.11-3.12 g/L), 454 for EtG (1.0-9739 ng/mL), 635 for PEth (0.014-6.0 µmol/L), 534 for PEth ≥ 0.050 µmol/L, and 315 for PEth ≥ 0.30 µmol/L. EtG and PEth concentrations seemed largely independent as the coefficient of determination (r²) between PEth and EtG concentrations was 0.15. However, when the EtG concentrations were evaluated for different subgroups depending on ethanol or PEth concentrations a statistically significant difference between successive higher concentrations was observed. EtG and PEth are independent measures of recent alcohol drinking reflecting different time windows. Their combined measurement in the same blood sample is possible and will provide valuable information regarding recent alcohol consumption as a complement to self-reporting.