Chronic Excessive Alcohol Consumption Research Articles

Fluorescence lifetime imaging of human pancreatic lipase activity using a novel probe for early diagnosis of severe acute pancreatitis

Severe Acute Pancreatitis, a serious condition caused by factors such as gallstones and chronic excessive alcohol consumption, with a very high mortality rate. Human pancreatic lipase (hPL) is a key digestive enzyme and abnormal activity levels of this enzyme are important indicators for diagnosing and monitoring pancreatic diseases. A fluorescent probe, LPP, has been developed to monitor the activity of hPL, especially in cases of SAP. The probe is based on cyanine isoindole derivatives, in vitro experiments confirmed the high specificity and sensitivity of the probe, with a detection limit of 0.012 U/mL, reactions completed within 10 min, and effective monitoring of pancreatic lipase activity in various biological samples. The stability and low cytotoxicity of LPP make it suitable for clinical applications, providing new tools and perspectives for the research and treatment of pancreatic diseases and related metabolic abnormalities. In addition, the change in fluorescence lifetime after the reaction of the probe with lipase allows for fluorescence lifetime imaging (FLIM), effectively monitoring the dynamic changes of hPL and enabling early diagnosis and monitoring of pancreatitis. This research not only enhances the understanding of pancreatic lipase activity detection but also has the potential to improve the diagnostics and treatment of pancreatitis.

Hepatoprotective effects and mechanisms of l-theanine and epigallocatechin gallate combined intervention in alcoholic fatty liver rats.

Chronic excessive alcohol consumption can lead to alcoholic fatty liver, posing substantial health risks. l-Theanine (LTA) and epigallocatechin gallate (EGCG) in tea exert antioxidant and hepatoprotective effects. However, the combined effects of LTA and EGCG on rats with alcoholic fatty liver, and the underlying mechanisms of such effects, remain unclear. In this study, Sprague Dawley (SD) rats were fed with alcohol for 6 weeks to induce alcoholic fatty liver. Subsequently, for another 6 weeks, the rats were administered LTA (200 mg kg-1 day-1), EGCG (200 mg kg-1 day-1), or a combination of LTA with EGCG (40 mg kg-1 day-1 l-Thea +160 mg kg-1 day-1 EGCG), respectively. The combined use of LTA and EGCG for alcoholic fatty liver disease had more significant effects than their individual administration. This combination reduced the activity of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) as well as the levels of hepatic triglyceride (TG), malondialdehyde (MDA), and reactive oxygen species (ROS) in the rats. The combined intervention also increased hepatic superoxide dismutase (SOD) and glutathione peroxidase activity. Reductions in hepatic fat accumulation and inflammatory responses were observed. The mechanism underlying these effects primarily involved the inhibition of fatty acid synthesis and the alleviation of lipid peroxidation through the downregulation of the mRNA and protein expression of TNF-α, SREBP1c, and CYP2E1 and the upregulation of the mRNA and protein expression of ADH1, ALDH2, Lipin-1, PPARαPPARα, AMPK, and PGC-1α, thereby promoting the oxidative decomposition of fatty acids and reducing the synthesis of cholesterol and glucose. l-Theanine and EGCG appear to be able to alleviate alcoholic fatty liver by modulating lipid metabolism and ameliorating oxidative stress, indicating their potential as natural active ingredients in anti-alcoholic fatty liver food products. © 2024 Society of Chemical Industry.

Exploring the Potential Role of Massa Medicata Fermentata in Alcoholic Liver Injury Disease Based on Network Pharmacology and Animal Experiments.

ALD is a chronic liver disease caused by chronic excessive alcohol consumption, for which there are no drugs with better efficacy. Ancient literature and modern studies have shown that Massa Medicata Fermentata (MMF) has a hangover effect and ameliorates hepatic inflammation, so we believe that MMF has a potential role in the treatment of alcoholic liver disease. UPLC-Q-Orbitrap HRMS was used to characterize the chemical constituents in MMF. The database was utilized to collect targets for the components and diseases, and cross-targeting analysis of the targets was performed. PPI, KEGG, GO enrichment analysis and molecular docking were performed using the core cross-targeting information to preliminarily validate the mechanism of action of MMF on disease. Finally, animal validation was carried out using male KM mice of the alcoholic liver injury model. MMF could play a role in the therapeutic prevention of alcoholic liver disease through the core targets AKT1, TNF, TP53, IL6 and CASP3 to regulate cancer pathways, lipid, and atherosclerosis, targeting IL-17 signaling, TNF signaling pathway, and hepatitis C, which was confirmed by animal pharmacodynamic experiments. This study serves as a rationale to support MMF in the treatment of ALD and meets the urgent need for clinical treatment of ALD. At the same time, it broadens the scope of clinical application of MMF.

Alcohol and Heart Failure

Alcohol is the most frequently consumed toxic substance in the world and remains a major global public health issue, with one in three adults consuming it worldwide. Alcohol use is a leading risk factor for disease, contributing to over 60 acute and chronic health conditions, with a particularly complex association with cardiovascular disease. Chronic excessive alcohol consumption is associated with a range of cardiac complications, including decreased myocardial contractility, hypertension, arrhythmias, MI and heart failure. However, low-level alcohol consumption is believed to have a protective effect against ischaemic heart disease and diabetes. In most cohort studies, small to moderate amounts of alcohol consumption have not been linked to heart failure, indicating a threshold effect of alcohol with individual (possibly genetic) predisposition rather than a continuous effect of exposure. This review article explores the potential benefits of alcohol on the heart, the association between alcohol use and alcoholic cardiomyopathy and the epidemiology, clinical correlates and management of alcoholic cardiomyopathy.

Proteomics and weighted gene correlated network analysis reveal glutamatergic synapse signaling in diazepam treatment of alcohol withdrawal.

Background: Alcohol use disorder (AUD) is characterized by chronic excessive alcohol consumption, often alternating with periods of abstinence known as alcohol withdrawal syndrome (AWS). Diazepam is the preferred benzodiazepine for treatment of alcohol withdrawal syndrome under most circumstances, but the specific mechanism underlying the treatment needs further research. Methods: We constructed an animal model of two-bottle choices and chronic intermittent ethanol exposure. LC-MS/MS proteomic analysis based on the label-free and intensity-based quantification approach was used to detect the protein profile of the whole brain. Weighted gene correlated network analysis was applied for scale-free network topology analysis. We established a protein-protein interaction network based on the Search Tool for the Retrieval of Interacting Genes (STRING) database and Cytoscape software and identified hub proteins by CytoHubba and MCODE plugins of Cytoscape. The online tool Targetscan identified miRNA-mRNA pair interactions. Results: Seven hub proteins (Dlg3, Dlg4, Shank3, Grin2b, Camk2b, Camk2a and Syngap1) were implicated in alcohol withdrawal syndrome or diazepam treatment. In enrichment analysis, glutamatergic synapses were considered the most important pathway related to alcohol use disorder. Decreased glutamatergic synapses were observed in the late stage of withdrawal, as a protective mechanism that attenuated withdrawal-induced excitotoxicity. Diazepam treatment during withdrawal increased glutamatergic synapses, alleviating withdrawal-induced synapse inhibition. Conclusion: Glutamatergic synapses are considered the most important pathway related to alcohol use disorder that may be a potential molecular target for new interventional strategies.

Daucosterol Alleviates Alcohol−Induced Hepatic Injury and Inflammation through P38/NF−κB/NLRP3 Inflammasome Pathway

Alcoholic liver disease (ALD) is caused by chronic excessive alcohol consumption, which leads to inflammation, oxidative stress, lipid accumulation, liver fibrosis/cirrhosis, and even liver cancer. However, there are currently no effective drugs for ALD. Herein, we report that a natural phytosterol Daucosterol (DAU) can effectively protect against liver injury caused by alcohol, which plays anti-inflammatory and antioxidative roles in many chronic inflammatory diseases. Our results demonstrate that DAU ameliorates liver inflammation induced by alcohol through p38/nuclear factor kappa B (NF-κB)/NOD-like receptor protein-3 (NLRP3) inflammasome pathway. Briefly, DAU decreases NF-κB nuclear translocation and inhibits NLRP3 activation by decreasing p38 phosphorylation. At the same time, DAU also protects against hepatic oxidative stress and lipid accumulation. In conclusion, our research provides a new clue about the protective effects of naturally active substances on ALD.

Added value evidences of a simultaneous determination of two phosphatidylethanol isoforms and ethylglucuronide in dried blood spots in forensic contexts

In clinical or postmortem forensic medicine, acute or chronical alcohol abuse are of main concern and direct/indirect biomarkers are widely use in order to assess recent or retrospective alcohol abuse. We aim to illustrate that a simultaneous determination of 3 directs ethanol biomarkers [two phosphatidylethanol isoforms, 16:0/18:1 (PEth18:1) and 16:0/20:4 (PEth20:4), and ethylglucuronide (EtGB)] using dried blood spot (DBS) can be a useful additional tool in many situations. (Case#1) Sexual assault of a 17-year-old girl with negative urine and blood ethanol concentration (BAC) (but urine EtG concentration was 10 mg/L) sampled 19 hours after the facts. (Case#2) Only blood sampling (negative BAC) was performed 14 hours after a 36-year-old man (with heavy psychiatric disorders) tried to kill 3 persons. He was reported drunk at the time of the attack by witnesses. (Case#3) A 27-year-old man was implicated in a car-related fatality that occurred 11 days before sampling. Blood and hair sampling were performed in order to investigate his alcoholic status: hair EtG concentration was < 5 pg/mg. (Case#4) The alcoholic status of a 43-year-old woman killed in a car accident had to be established. BAC was 140 mg/L in the postmortem peripheral blood (only available biological sample; sampled less than one hour after death and kept refrigered until time of analysis, 2 days later). (Case#5) A low BAC (70 mg/L) was found in postmortem cardiac blood (only sample available) sampled on the corps of a 42-year-old man found submerged in the sea. (Case#6) Peripheral blood sampling (for BAC determination) and direct DBS sampling (MitraTM) were performed at the time of discovery of the 36-year-old man corpse, allegedly heavy drinker according to the testimonies (BAC was 2810 mg/L). Blood samples can alternatively consist in 10 μL from usual blood sampling tubes (e.g. fluoride preserved) transferred on DBS card, or direct DBS sampling using Volumetric Absorptive MicroSampling devices (VAMS; HemaxisTM, MitraTM). These DBS are analyzed using a LC-MS/MS (routinely used in clinical toxicology) method (LLOQ: 2 μg/L for the 3 analytes). Results (μg/L; PEth18:1, PEth20:4 and EtGB, respectively) were: – (Case#1) 22, 2 and 300, in coherence with both a recent alcohol intake hypothesis [EtGB above the 45 μg/L threshold supported an alcohol consumption [> 2 standard units (SU)] in the last 24 to 48 hours] and that this girl was not an excessive drinker [owing to the low PEths concentrations]; – (Case#2) 98, 103 and 71, consistent with an alcohol intake hypothesis arround the time of the attack by this man who had a significant consumption of alcohol in the last 10 days as PEth 20:4 concentration was similar to the PEth18:1 one; – (Case#3) 9, < 2 and < 2, and did not contradict the negative EtG hair result as these low values were, at most, compatible with an “occasional” or “social drinker” status (< 14 SU per week); – (Case#4) 817, 44 and 130. EtGB positive result supported a recent alcohol consumption and the very high PEth18:1 value strongly suggested chronic excessive alcohol consumption; – (Case#5) negative (< 2 μg/L) for the 3 analytes in postmortem cardiac blood, in agreement with both the hypothesis of a light postmortem ethanol production, and a probable “abstinent” or “occasional consumer” status of the victim; – (Case#6) 3470, 1030 and 13,400, supporting the massive alcohol intoxication hypothesis close to the death of a heavy drinker. This analytical tool is of high interest when only blood samples are available, although postmortem use (due to PEths neoformation issues) should be limited to cases with negative BAC, or when blood sampling is performed close to death using a VAMS or a refrigerated preserved (< 4 days) tube.

Reply.

We appreciate the opportunity to respond to Mondia et al. This comment raises concerns about preterm birth analysis, which comprised a subgroup of our study population. First, Mondia et al. point out the lack of information on hospital admission and duration in the neonatal period. The Constances cohort included adult participants ≥18 years old. Birth data were retrospectively collected from health booklets. Indeed, information about neonatal hospitalization information is not always detailed in health booklets and therefore was not collected for the cohort. Second, regarding breastfeeding and early weight catch‐up, there were missing data in our sample. We are currently performing a data cleaning for these variables, but we expect availability for only a subset of the cohort. We agree with Mondia et al. that this data analysis would improve the comprehension of the impact of preterm birth and low birth weight on future health. Finally, congenital or genetic disorders (not accounted for in this analysis) represent a minor cause of prematurity/low birth weight. Nevertheless, we would like to correct the comment regarding metabolic disorders given that we did account for metabolic syndrome in participants through a mediation analysis. Finally, liver biopsy was not considered in this general purposed and observational cohort. However, common causes of liver fibrosis (other than NAFLD) were systematically searched for through a physician‐administrated questionnaire. People with chronic viral hepatitis or excessive alcohol consumption (i.e., >20 and >30 g per day for women and men, respectively) were excluded from our analyses.

Targeting Unmet Clinical Needs in the Treatment of Alcohol Use Disorder.

Alcohol Use Disorder (AUD) is a chronic psychiatric disorder marked by impaired control over drinking behavior that poses a significant challenge to the individual, their community, the healthcare system and economy. While the negative consequences of chronic excessive alcohol consumption are well-documented, effective treatment for AUD and alcohol-associated diseases remains challenging. Cognitive and behavioral treatment, with or without pharmaceutical interventions, remain the most commonly used methods; however, their efficacy is limited. The development of new treatment protocols for AUD is challenged by difficulty in accurately measuring patterns of alcohol consumption in AUD patients, a lack of a clear understanding of the neuropsychological basis of the disorder, the high likelihood of AUD patients relapsing after receiving treatment, and the numerous end-organ comorbidities associated with excessive alcohol use. Identification and prediction of patients who may respond well to a certain treatment mechanism as well as clinical measurement of a patient's alcohol exposure are bottlenecks in AUD research which should be further addressed. In addition, greater focus must be placed on the development of novel strategies of drug design aimed at targeting the integrated neural pathways implicated in AUD pathogenesis, so that next-generation AUD treatment protocols can address the broad and systemic effects of AUD and its comorbid conditions.

Association between birth weight, preterm birth, and nonalcoholic fatty liver disease in a community-based cohort.

The association between birth weight (BW) and metabolic outcomes has been described since the 1980s but NAFLD has been rarely studied. This study aimed to investigate the association between BW and NAFLD occurrence in adult subjects. The study population consisted of participants from the French nationwide Constances cohort from 2012 to 2019. Participants with a history of chronic viral hepatitis or excessive alcohol consumption were excluded. Noninvasive diagnosis of NAFLD and fibrosis was performed using a combination of the Fatty Liver Index (FLI) and the Forns Index. The relationship between BW and NAFLD was analyzed with a sex-stratified logistic regression model adjusted for sociodemographic parameters, lifestyle, and birth term, whereas liver fibrosis was analyzed with a sex-stratified linear regression model. In total, 55,034 individuals with reliable BW were included (43% men, mean age: 38 years). NAFLD (FLI≥60) was present in 5530 individuals (10%). Multivariate logistic regression showed a significant U-shaped relationship between BW and NAFLD, with no significant interaction with sex. A significant and slightly decreasing association was found between BW and Forns Index (β=-0.05; p=0.04). Premature birth (OR, 1.23; 95% CI, 1.03-1.48 for birth between 33 and 37 weeks versus≥37 weeks) was associated with NAFLD, with a significant direct effect of premature birth, and without an indirect effect of low BW in mediation analysis. Forns Index was not significantly higher in participants with preterm birth compared to full-term birth. This large prospective adult-based cohort confirms the relationship between BW and NAFLD occurrence.

Determination of phosphatidyl ethanol (PEth) 16:0/18:1 in dried blood samples of drivers involved in traffic accidents: A pilot study

In routine forensic toxicology practices, blood alcohol concentration (BAC) levels are measured in traffic accidents that ended up in emergency departments. Nevertheless, since the elimination of ethanol from the blood is fast and the detection time is short, BAC cannot indicate the occurrence of chronic excessive alcohol consumption. Phosphatidylethanol (PEth) is a unique ethanol direct biomarker that occurs only in the presence of phospholipase D enzyme in erythrocyte membranes during alcohol intake, and it indicates alcohol intake.In this study, both whole blood and dried blood samples were collected from 50 patients who were admitted to Cukurova University Hospital Emergency Department due to a traffic accident. While studying BAC in whole blood samples, PEth 16:0/18:1 analysis was performed on dried blood samples by LC-MS/MS.According to the BAC (50 mg/dL) value, the legal limit in Turkey, the optimal threshold PEth 16:0/18:1 value was set as 160 ng/mL and over. This study determined that 15 people with above PEth 16:0/18:1 concentrations above 160 ng/mL were classified as excessive alcohol consumption.The data obtained in this study showed a positive correlation between BAC and PEth concentration when driving under the influence of ethanol.

Associations between lead, cadmium, mercury, and arsenic exposure and alanine aminotransferase elevation in the general adult population: an exposure-response analysis.

Cadmium, lead, mercury, and arsenic are among the most toxic environmental contaminants. Serum alanine aminotransferase (ALT) is the most common liver biomarker. This analysis aimed to explore the associations between blood cadmium, lead, mercury, urinary total arsenic, and dimethylarsinic acid and ALT elevation in adults. Data were extracted from 5 National Health and Nutrition Examination Survey cycles (NHANES) 2007-2016. Patients with chronic viral hepatitis and excessive alcohol consumption were excluded. ALT elevation was defined according to the 2017 American College of Gastroenterology Clinical Guideline. Logistic models and restricted cubic splines were adopted to assess the exposure-response relationships. Comparing the highest to lowest quintile of exposure, the multivariable-adjusted odds ratios (95% confidence intervals) of ALT elevation were 1.38 (1.07-1.78) for blood lead (Pfor trend = 0.01), 1.37 (1.16-1.62) for blood mercury (Pfor trend < 0.01), 0.94 (0.78-1.14) for blood cadmium (Pfor trend = 0.64), 1.07 (0.79-1.45) for urinary total arsenic (Pfor trend = 0.81), and 1.25 (0.94-1.66) for urinary dimethylarsinic acid (Pfor trend = 0.18). The associations between blood lead and mercury and ALT elevation were only observed in women. In addition, the associations between urinary total arsenic [1.53 (1.02-2.29), Pfor trend = 0.02] and dimethylarsinic acid [2.17 (1.05-4.49), Pfor trend = 0.02] and ALT elevation were also observed in women. Dose-response analysis showed that there was no safe exposure threshold of blood lead and mercury's toxic effect on ALT elevation, respectively. In conclusion, lead, mercury and arsenic were associated with ALT elevation in adults, and the associations were mainly observed in women.

MicroRNA-29b ameliorates hepatic inflammation via suppression of STAT3 in alcohol-associated liver disease.

Alcohol-associated liver disease (ALD) is induced by chronic excessive alcohol consumption resulting in the clinical manifestations of steatosis, inflammation, and cirrhosis. MicroRNA-29b (miR-29b) is mainly expressed in hepatic nonparenchymal cells, and its expression level varies in different diseases. In this study, we aimed to determine the role of miR-29b in a mouse model of alcohol-associated liver disease. Wild-type (WT) and miR-29b knockout (miR-29b-/-) mice were fed a Lieber-DeCarli liquid diet containing 5% alcohol for 10 days, followed by gavage of a single dose of ethanol (5g/kg body weight). Histology, immunoblotting, and biochemical analyses were then conducted for comparison. miR-29b expression was decreased in the livers of chronic-plus-binge ethanol-fed mice. Further analysis revealed that alcohol exposure exacerbated hepatic injury by significantly increasing serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, with decreased survival rates for miR-29b-/- mice. Results from the luciferase assay indicated that miR-29b negatively regulated the signal transducer and activator of transcription 3 (STAT3). Depletion of miR-29b led to an increase in STAT3 and more noticeable inflammation in the liver, whereas overexpression of miR-29b downregulated STAT3 and proinflammatory cytokine expression in primary mouse peritoneal macrophages. Taken together, these results demonstrate a novel association between miR-29b and ALD. miR-29b plays a hepatoprotective role in alcohol-induced inflammation and liver injury by targeting STAT3.

Therapeutic Effects of Myriocin in Experimental Alcohol-Related Neurobehavioral Dysfunction and Frontal Lobe White Matter Biochemical Pathology.

Chronic excessive alcohol consumption causes white matter degeneration with myelin loss and impaired neuronal conductivity. Subsequent rarefaction of myelin accounts for the sustained deficits in cognition, learning, and memory. Correspondingly, chronic heavy or repeated binge alcohol exposures in humans and experimental models alter myelin lipid composition leading to build-up of ceramides which can be neurotoxic and broadly inhibitory to brain functions. This study examined the effects of chronic + binge alcohol exposures (8 weeks) and intervention with myriocin, a ceramide inhibitor, on neurobehavioral functions (Open Field, Novel Object Recognition, and Morris Water Maze tests) and frontal lobe white matter myelin lipid biochemical pathology in an adult Long-Evans rat model. The ethanol-exposed group had significant deficits in executive functions with increased indices of anxiety and impairments in spatial learning acquisition. Myriocin partially remediated these effects of ethanol while not impacting behavior in the control group. Ethanol-fed rats had significantly smaller brains with broadly reduced expression of sulfatides and reduced expression of two of the three sphingomyelins detected in frontal white matter. Myriocin partially resolved these effects corresponding with improvements in neurobehavioral function. Therapeutic strategies that support cerebral white matter myelin expression of sulfatide and sphingomyelin may help remediate cognitive-behavioral dysfunction following chronic heavy alcohol consumption in humans.

Chronic Alcohol Consumption and its Impact on Bone and Metabolic Health - A Narrative Review.

Chronic consumption of a large quantity of alcohol often results in the disruption of the communication between the nervous, endocrine and immune systems leading to profound and serious consequences at the physiological and behavioral levels. The overall impact of excessive alcohol consumption on bone health, metabolic profile and body composition, especially at moderate levels, is not well understood. Chronic excessive alcohol consumption adversely affects bone health through multiple mechanisms leading to low bone mass. It may also be significantly associated with various components on the metabolic syndrome. This review summarizes the findings from published studies that provide consistent evidence on the various effects of alcohol abuse on the bone health and metabolism.

Investigating the use of PEth, CDT and MCV to evaluate alcohol consumption in a cohort of homeless individuals– A comparison of different alcohol biomarkers

In a cohort including individuals with suspected high alcohol consumption, the concentrations of the indirect alcohol biomarkers carbohydrate-deficient transferrin (CDT) and mean corpuscular volume (MCV) and the direct alcohol biomarker phosphatidylethanol (PEth) were investigated. Blood alcohol concentration (BAC) was analysed as a marker for acute alcohol ingestion.In addition to questions about subjective alcohol consumption behaviour, 147 homeless persons underwent a physical examination with blood sampling. BAC, PEth, CDT and MCV were determined in the blood samples. Special focus was on the comparison of PEth and CDT for indicating excessive alcohol consumption.BAC was measured above 0.1‰ in 39 blood samples (0.1–2.5‰, median 0.75‰). PEth was detected in all of them. Overall, PEth was positive (≥10 ng/ml) in 104 samples (71%) (11–5687 ng/ml, median 650 ng/ml) with 68 (46%) being above the cut-off for excessive alcohol consumption (210 ng/ml). In 26 subjects PEth was the only positive alcohol biomarker. CDT was ≥ 1.7% in 66 cases (47%) (1.8–22.2%, median 4.4%) and ≥ 2.5% in 52 (35%) cases. MCV was elevated (≥95 fl) in 58 subjects (39%). CDT and PEth concentrations showed a significant positive correlation (spearman's correlation coefficient ρ = 0.77, p < 0.001). PEth concentrations were significantly higher in samples that were also CDT positive than solely PEth positive (p = 0.004). PEth did not indicate excessive alcohol consumption (< 210 ng/ml) in eight and two cases in which CDT was ≥ 1.7% and ≥ 2.5%, respectively. On the other hand, CDT was< 1.7% and< 2.5% in ten and 18 cases, respectively, in which PEth was above cut-off for excessive alcohol consumption. Taking the self-reports of the participants into consideration, PEth’s sensitivity for detecting excessive alcohol consumption was 100% (10 ng/ml) and 94% (210 ng/ml) and CDT’s was 88% (1.7%) and 75% (2.5%).In individuals of the investigated cohort unusually high concentrations of the alcohol consumption markers PEth and CDT were quantified, which proves the assumption of chronic excessive alcohol consumption in parts of the cohort. PEth was the marker that was positive most often and was more sensitive for excessive alcohol consumption than CDT.

Antisense-induced downregulation of major circadian genes modulates the expression of histone deacetylase-2 (HDAC-2) and CREB-binding protein (CBP) in the medial shell region of nucleus accumbens of mice exposed to chronic excessive alcohol consumption.

Circadian genes in the medial accumbal shell (mNAcSh) region regulate binge alcohol consumption. Here, we investigated if antisense-induced knockdown of major circadian genes (Per1, Per2, and NPAS2) in the mNAcSh of mice exposed to intermittent access two-bottle choice (IA2BC) paradigm modulates the expression of histone deacetylase-2 (HDAC-2) and CREB-binding protein (CBP), key epigenetic modifiers associated with withdrawal-associated behaviors such as anxiety. Adult male C57BL/6J mice (N=28), surgically implanted with bilateral guide cannulas above the mNAcSh, were chronically (4weeks) exposed to alcohol (20% v/v) or saccharin (0.03%) via IA2BC paradigm. In the fourth week, a mixture of antisense (AS-ODNs; N=14/group) or nonsense (NS-ODNs; N=14/group) oligodeoxynucleotides against circadian genes were bilaterally infused into the mNAcSh. Subsequently, alcohol/saccharin consumption and preference were measured followed by euthanization of animals and verification of microinjection sites by visual inspection and the expression of HDAC-2 and CBP by using RT-PCR along with the verification of antisense-induced downregulation of circadian genes in the mNAcSh. As compared with NS-ODNs, AS-ODNs infusion significantly attenuated the alcohol-induced increase in HDAC-2 and reduction in CBP expression in the mNAcSh along with a significant reduction in alcohol consumption and preference. No significant effect was observed on either saccharin consumption or preference. Our results suggest that circadian genes in the mNAcSh may have a causal to play in mediating epigenetic changes observed after chronic alcohol consumption.

Clinical Significance of Gamma-Glutamyltranspeptidase Combined with Carbohydrate-Deficient Transferrin for the Assessment of Excessive Alcohol Consumption in Patients with Alcoholic Cirrhosis.

Background: This study aimed to compare the diagnostic performance of carbohydrate-deficient transferrin (CDT) and gamma-glutamyltranspeptidase (γ-GTP) to assess the single and combined benefits of these biological markers for the detection of chronic excessive alcohol consumption in patients with alcoholic cirrhosis. Methods: Biological markers were determined in blood samples from patients with alcoholic cirrhosis (drinking group, n = 35; nondrinking group, n = 81). The prediction accuracy of %CDT alone, γ-GTP alone, and their combination for the detection of excessive alcohol consumption was determined in patients with alcoholic cirrhosis. Results: Serum total bilirubin, alanine aminotransferase, aspartate aminotransferase, γ-GTP, and alkaline phosphatase levels and %CDT were significantly higher and serum albumin levels were significantly lower in the drinking group than in the nondrinking group. The combination of %CDT and γ-GTP compared with %CDT or γ-GTP alone showed a higher prediction accuracy. The combination of %CDT and γ-GTP exhibited a higher specificity than γ-GTP alone. However, in terms of sensitivity, no significant difference was found between single or combined markers. Conclusions: The combination of %CDT and γ-GTP is considered a useful biomarker of chronic excessive alcohol consumption in patients with alcoholic cirrhosis.

Coronavirus lockdown: Excessive alcohol consumption and illicit substance use in DUI subjects

Objective This study investigates the consequences of the SarS-CoV-2 outbreak and of the resulting control measures on alcohol and illicit substance use in a high-risk population for substance-related disorders, utilizing an integrated medico-legal and toxicological approach. Methods The research was structured as a retrospective case-control study of subjects found to be driving under the influence (DUI) of alcohol and/or other psychoactive substances who were examined for driver’s license regranting. Alcohol and/or drug use was assessed by comparing cases examined in the period from May to August 2020 (immediately after the lockdown in Italy) to control subjects examined in the same period in 2019. DUI subjects were examined by an integrated approach, descriptive analyses were conducted, and significance was determined by chi-square and Mann-Whitney tests. Variables linked to the pandemic outbreak and resulting lockdown were investigated as predictive factors in determining unfitness to drive. Results Cases (281) were significantly different from controls (261) concerning the judgment of unfitness to drive (p<.001) and had more subjects with chronic excessive alcohol use and/or illicit substance use. The two groups were rather homogeneous concerning the other variables, except for a difference in blood alcohol concentration (BAC) at the time of DUI (p = .027). No statistical association was found between the investigated variables linked to the lockdown and the judgment of unfitness to drive. Conclusions Chronic excessive alcohol consumption and illicit substance use were more frequently observed in cases, which suggests a possible correlation between the pandemic/lockdown restrictions and an increase in psychoactive substance misuse. While these potentially correlative factors are discussed in this article, they require further study. If confirmed, the results should be considered in forensic and clinical settings.

Metabolic Profiling Analysis Reveals the Potential Contribution of Barley Sprouts against Oxidative Stress and Related Liver Cell Damage in Habitual Alcohol Drinkers.

Chronic excessive alcohol consumption is associated with multiple liver defects, such as steatosis and cirrhosis, mainly attributable to excessive reactive oxygen species (ROS) production. Barley sprouts (Hordeum vulgare L.) contain high levels of polyphenols that may serve as potential antioxidants. This study aimed to investigate whether barley sprouts extract powder (BSE) relieves alcohol-induced oxidative stress and related hepatic damages in habitual alcohol drinkers with fatty liver. In a 12-week randomized controlled trial with two arms (placebo or 480 mg/day BSE; n = 76), we measured clinical markers and metabolites at the baseline and endpoint to understand the complex molecular mechanisms. BSE supplementation reduced the magnitude of ROS generation and lipid peroxidation and improved the glutathione antioxidant system. Subsequent metabolomic analysis identified alterations in glutathione metabolism, amino acid metabolism, and fatty acid synthesis pathways, confirming the role of BSE in glutathione-related lipid metabolism. Finally, the unsupervised machine learning algorithm indicated that subjects with lower glutathione reductase at the baseline were responders for liver fat content, and those with higher fatigue and lipid oxidation were responders for γ-glutamyl transferase. These findings suggest that BSE administration may protect against hepatic injury by reducing oxidative stress and changing the metabolism in habitual alcohol drinkers with fatty liver.