Determination of cortisol in hair using UHPLC–MS/MS: application to patients admitted for ethanol dependence treatment

Aim: Cortisol hair levels can be used to evaluate chronic stress status. In this context, an improved UHPLC–MS/MS assay for the determination of cortisol in hair was developed and validated. Materials & methods: Hair was extracted with methanol for 4 h at 25°C. Chromatographic run time was 5.5 min. The assay was linear in the range of 1–250 pg mg-1. Precision was 3.6–12.2% and accuracy 97.1–103.8%. The method was applied in hair from 19 volunteers admitted at a rehabilitation clinic, with ethanol consumption classified using ethyl glucuronide hair levels. Conclusion: Abstinent/chronic moderate ethanol consumers had significantly lower cortisol hair levels than chronic excessive consumers. This is the first study evaluating cortisol hair levels in ethanol abuse patients using an objective marker for ethanol consumption.

Essential Thrombocytosis and Hepatocellular Carcinoma: A Mechanistic Challenge

Introduction/Objective Thrombocytosis has been reported in 8.2% of patients with Hepatocellular carcinoma [HCC] and has been attributed to increased Thrombopoietin [TPO] production by the tumor. Methods/Case Report We present a case of a 68-year-old male with a history of Hepatitis C and ethanol abuse presented with complaints of headache and chills. Imaging studies revealed a large mass in the liver [right lobe] suggesting an HCC of fibrolamellar type. His platelet count [PC] was 965 K/mcL. The serum AFP [Alpha feto protein] concentration was >51,800 ng/ml. There was no evidence of metastatic disease and the patient was started on Capecitabine and radiation therapy targeting the liver mass. Lung metastasis became evident and Sorafenib was added. His PCs and AFP concentrations however continued decreasing to 306 K/mcL and 460.7 ng/ml respectively. His PC then increased to a peak of 1.32 million/mcL 10 months later. The molecular workup done at our reference lab was positive for CALR [Calreticulin] mutation. The patient was started on Hydroxyurea and his PC decreased to 323 K/mcL at the time of writing. His AFP has remained stable. This case illuminates the complicated relationship between HCC and thrombocytosis. The therapy directed against the HCC did decrease PC and AFP concentration in our case. The second wave of thrombocytosis led to the discovery of CALR mutation, and the patient was diagnosed with Essential Thrombocytosis [ET]. Treatment with Hydroxyurea again decreased the PC while AFP concentration was increased but stable. Thrombopoietin levels have remained stable for the patient after treatment with Hydroxyurea. Results (if a Case Study enter NA) NA Conclusion Although the question of which mechanism(or possibly both) was at work in this case cannot be resolved definitely, two points are worth noting. The first is that- the assumption that the thrombocytosis was related to the HCC delayed testing for mutations associated with Essential Thrombocytosis. The second is that the TPO assay that may have yielded useful information in sorting out the alternatives was not ordered early enough before starting hydroxyurea.

Injury caused by alcoholic cardiomyopathy in spontaneous ethanol drinking rats

ABSTRACTWhen speaking of pathologies caused or aggravated by the constant ingestion of ethanol, people with liver and central nervous system diseases soon come to mind, however, the acute intake of large amounts of ethanol and chronic abuse induce toxic effects in the majority of tissues. The heart is highlighted, since alcoholic cardiomyopathy (AC) has prevalence among alcoholics of 23 to 40% and occurs more frequently in men than in women. AC is characterized by dilation and poor contraction of one or both ventricles in the presence of increased ventricular wall thickness, along with a long history of ethanol abuse and no other cause identified. Our aim is to quantify the rate of cardiac tissue replacement, collagen fiber deposition and pro inflammatory cytokines in the left ventricle myocardium of volunteer ethanol drinking rats.

Increased dopaminergic neurotransmission results in ethanol dependent sedative behaviors in Caenorhabditis elegans

Ethanol is a widely used drug, excessive consumption of which could lead to medical conditions with diverse symptoms. Ethanol abuse causes dysfunction of memory, attention, speech and locomotion across species. Dopamine signaling plays an essential role in ethanol dependent behaviors in animals ranging from C. elegans to humans. We devised an ethanol dependent assay in which mutants in the dopamine autoreceptor, dop-2, displayed a unique sedative locomotory behavior causing the animals to move in circles while dragging the posterior half of their body. Here, we identify the posterior dopaminergic sensory neuron as being essential to modulate this behavior. We further demonstrate that in dop-2 mutants, ethanol exposure increases dopamine secretion and functions in a DVA interneuron dependent manner. DVA releases the neuropeptide NLP-12 that is known to function through cholinergic motor neurons and affect movement. Thus, DOP-2 modulates dopamine levels at the synapse and regulates alcohol induced movement through NLP-12.

Biochemical and Behavioral Consequences of Ethanol Intake in a Mouse Model of Metabolic Syndrome

Ethanol abuse is a common issue in individuals with sedentary lifestyles, unbalanced diets, and metabolic syndrome. Both ethanol abuse and metabolic syndrome have negative impacts on the central nervous system, with effects including cognitive impairment and brain oxidative status deterioration. The combined effects of ethanol abuse and metabolic syndrome at a central level have not yet been elucidated in detail. Thus, this work aims to determine the effects of ethanol intake on a mouse model of metabolic syndrome at the behavioral and biochemical levels. Seven-week-old male control (B6.V-Lep ob/+JRj) and leptin-deficient (metabolic syndrome) (B6.V-Lep ob/obJRj) mice were used in the study. Animals were divided into four groups: control, ethanol, obese, and obese–ethanol. Ethanol consumption was monitored for 6 weeks. Basal glycemia, insulin, and glucose overload tests were performed. To assess short- and long-term memory, an object recognition test was used. In order to assess oxidative status in mouse brain samples, antioxidant enzyme activity was analyzed with regard to glutathione peroxidase, glutathione reductase, glutathione, glutathione disulfide, lipid peroxidation products, and malondialdehyde. Ethanol intake modulated the insulin response and impaired the oxidative status in the ob mouse brain.

Deleterious Effects of Ethanol, Δ(9)-Tetrahydrocannabinol (THC), and Their Combination on the Spatial Memory and Cognitive Flexibility in Adolescent and Adult Male Rats in the Barnes Maze Task

Research demonstrates that adolescents differ from adults in their response to drugs of abuse. The aim of the present study was to examine the influence of ethanol, Δ9-tetrahydrocannabinol hydrochloride (THC), and a combination of these drugs given during adolescence on spatial memory in adolescent and adult rats. Thus, adolescent rats (postnatal day (PND) 30) were subjected to the following groups: 0.9% NaCl; 1.5 g/kg ethanol; 1.0 mg/kg THC; 1.5 g/kg ethanol + 1.0 mg/kg THC. Rats received drug injection four times at three-day intervals. One day after the last injection, half of the treated animals were tested in the Barnes maze task, whereas the remaining animals were tested on PND 70. Results show that there was a significant age effect on spatial memory in the Barnes maze task after these drug administrations. Adolescent animals demonstrated more potent deficits in the spatial learning and memory (probe trial) and in cognitive flexibility (reversal learning) than did adults. However, in adult rats that received these drugs in adolescence, memory decline was observed only after ethanol and ethanol + THC administration. Thus, our results are important in understanding the deleterious impact of THC and/or ethanol abuse during adolescence on memory function across the lifespan (adolescent versus adult).

Trends of Hospitalization for Acute Alcohol Intoxication in Slovenian Children and Adolescents with and without Dual Disorder. Implications for a Correct Intervention

Background: Binge drinking and other forms of ethanol abuse are, when present, a serious problem in preteens and adolescents worldwide. Aim: The present study has analyzed the trend in alcohol-related intoxications requiring the hospitalization of children, adolescents and young adults aged less than 21 years in Slovenia in the 1999–2018 period. Methods: We performed a retrospective study on patients discharged after hospitalizations due to mental and behavioral disorders due to acute alcohol intoxication (MBDAAI) or hospitalizations due to the toxic effects of alcohol (TEA We considered three groups: children (aged 10–14), adolescents (aged 15–19) and young adults (20–21 years old). Hospitalization rates and time trends were analyzed using joinpoint regression to obtain the annually calculated age- and sex-specific rates and the annual percentage of change (APC). Results: Considering a total of 2912 MBDAAI-hospitalizations, 15–19-year-old subjects showed a significantly higher hospitalization rate compared to the immediately younger and older age groups and a significant increase in hospitalization rates in the period 1999–2011, followed by a significant decrease. Considering 1143 TEA-hospitalizations, we observed a continuous decrease in the hospitalization rates for children and young adults and, conversely, a continuous even if less than significant increase for adolescents aged 15–19. Conclusions: Alcohol consumption in Slovenian children and adolescents is a highly important health concern. Special attention to public health problem of severe alcohol abuse requiring hospitalization in children and adolescents is needed, especially with possible crisis of SARS-CoV-2/Covid-19 situation.

Extensive Superior Vena Caval Territory Thrombosis and Pulmonary Embolism: A Rare Clinical Entity of Haemorrhagic Pancreatitis

Acute haemorrhagic pancreatitis is a severe form of pancreatitis often encountered in ethanol abuse. Extensive venous thrombosis resulting in pulmonary embolism is a rare presenting clinical entity of acute haemorrhagic pancreatitis. Here, we report a young male with an extensive deep vein thrombosis involving superior vena caval territory associated with haemorrhagic pancreatitis presented with pulmonary embolism managed supportively. Prompt recognition and appropriate intervention of this rare complication would improve the outcome in patients with acute pancreatitis.

NOS-2 Participates in the Behavioral Effects of Ethanol Withdrawal in Zebrafish

Nitric oxide has been implicated in symptoms of ethanol withdrawal in animal models. Zebrafish have been used as models to study neurobehavioral effects of ethanol (EtOH) withdrawal, but the mechanisms associated with these effects are not yet clear. Adult zebrafish were treated with 1% EtOH for 20 min per day for 8 days, injected with the nitric oxide synthase 2 (NOS-2) inhibitor aminoguanidine (50 mg/kg), and allowed to experience withdrawal (WD) in their hometanks for 7 days. EtOH WD increased anxiety-like behavior in the novel tank test, an effect that was blocked by aminoguanidine. EtOH WD also increased brain levels of nitrite, an effect that was partially blocked by aminoguanidine. These results underline a novel mechanism by which NOS-2 controls anxiety-like responses to ethanol withdrawal, with implications for the mechanistic study of symptoms associated with chronic ethanol abuse.