Chronic Alcohol Treatment Research Articles

Anticancer Effects of New Disulfiram Analogs.

Disulfiram (DSF), an irreversible aldehyde dehydrogenase 2 (ALDH2) inhibitor, is an U.S. Food and Drug Administration (FDA)-approved drug for the treatment of chronic alcoholism. Recent studies have reported an interesting antitumor activity of DSF against a wide range of human malignancies, while a growing number of ongoing and completed clinical trials have provided evidence supporting the repurposing of DSF as an anticancer treatment. Nevertheless, despite its current clinical indications and potential future therapeutic applications for treating various diseases, DSF is associated with serious side effects attributed to the inhibition of ALDH2. We have recently synthesized DSF analogs (2a-v) with limited inhibition of ALDH2. Here, we report the anticancer activity of these molecules by highlighting their effects on cell signaling in Jurkat cells. DSF and two DSF analogs, 2g and 2r, all stimulated apoptotic signaling pathways, although the 2g analog activated more apoptosis-related genes and induced higher levels of apoptosis in vitro. Differential gene expression data suggested that compounds 2g and 2r specifically reprogram target cells to downregulate pathways related to cell growth and division, while upregulating pathways related to apoptosis or differentiation. Interestingly, both compounds 2g and 2r had more differentially expressed genes related to DNA damage pathways (including those related to apoptosis) when compared to DSF, which may offer insights into their mechanisms of action.

Toll-like receptor 7: A novel neuroimmune target to reduce excessive alcohol consumption

Toll-like receptors (TLRs) are a family of innate immune receptors that recognize molecular patterns in foreign pathogens and intrinsic danger/damage signals from cells. TLR7 is a nucleic acid sensing endosomal TLR that is activated by single-stranded RNAs from microbes or by small noncoding RNAs that act as endogenous ligands. TLR7 signals through the MyD88 adaptor protein and activates the transcription factor interferon regulatory factor 7 (IRF7). TLR7 is found throughout the brain and is highly expressed in microglia, the main immune cells of the brain that have also been implicated in alcohol drinking in mice. Upregulation of TLR7 mRNA and protein has been identified in postmortem hippocampus and cortex from AUD subjects that correlated positively with lifetime consumption of alcohol. Similarly, Tlr7 and downstream signaling genes were upregulated in rat hippocampal and cortical slice cultures after chronic alcohol exposure and in these regions after chronic binge-like alcohol treatment in mice. In addition, repeated administration of the synthetic TLR7 agonists imiquimod (R837) or resiquimod (R848) increased voluntary alcohol drinking in different rodent models and produced sustained upregulation of IRF7 in the brain. These findings suggest that chronic TLR7 activation may drive excessive alcohol drinking. In the brain, this could occur through increased levels of endogenous TLR7 activators, like microRNAs and Y RNAs. This review explores chronic TLR7 activation as a pathway of dysregulated neuroimmune signaling in AUD and the endogenous small RNA ligands in the brain that could perpetuate innate immune responses and escalate alcohol drinking.

Antimicrobial Activity of N,N-Diethyldithiocarbamate against Ureaplasma parvum and Ureaplasma urealyticum.

Ureaplasma species (Ureaplasma spp.) are commonly found as commensals in the human urogenital tracts, although their overgrowth can lead to infection in the urogenital tract and at distal sites. Furthermore, ureaplasmas lack a cell wall and do not synthesize folic acid, which causes all β-lactam and glycopeptide antibiotics, and sulfonamides and diaminopyrimidines, to be of no value. The antibiotics used in therapy belong to the fluoroquinolone, tetracycline, chloramphenicol and macrolide classes. However, the growing incidence of antibiotic-resistant Ureaplasma spp. in the population becomes a problem. Thus, there is a need to search for new drugs effective against these bacteria. Since 1951, the FDA-approved, well-tolerated, inexpensive, orally administered drug disulfiram (DSF) has been used in the treatment of chronic alcoholism, but recently, its antimicrobial effects have been demonstrated. The main biological metabolite of DSF, i.e., N,N-diethyldithiocarbamate (DDC), is generally believed to be responsible for most of the observed pharmacological effects of DSF. In the presented studies, the effect of DDC at concentrations of 2 µg/mL, 20 µg/mL and 200 µg/mL on the growth and survival of Ureaplasma urealyticum and Ureaplasma parvum was tested for the first time. The results indicated that all the used DDC concentrations showed both bacteriostatic and bactericidal activity against both tested strains.

Repurposing disulfiram, an alcohol-abuse drug, in neuroblastoma causes KAT2A downregulation and in vivo activity with a water/oil emulsion

Neuroblastoma, the most common type of pediatric extracranial solid tumor, causes 10% of childhood cancer deaths. Despite intensive multimodal treatment, the outcomes of high-risk neuroblastoma remain poor. We urgently need to develop new therapies with safe long-term toxicity profiles for rapid testing in clinical trials. Drug repurposing is a promising approach to meet these needs. Here, we investigated disulfiram, a safe and successful chronic alcoholism treatment with known anticancer and epigenetic effects. Disulfiram efficiently induced cell cycle arrest and decreased the viability of six human neuroblastoma cell lines at half-maximal inhibitory concentrations up to 20 times lower than its peak clinical plasma level in patients treated for chronic alcoholism. Disulfiram shifted neuroblastoma transcriptome, decreasing MYCN levels and activating neuronal differentiation. Consistently, disulfiram significantly reduced the protein level of lysine acetyltransferase 2A (KAT2A), drastically reducing acetylation of its target residues on histone H3. To investigate disulfiram’s anticancer effects in an in vivo model of high-risk neuroblastoma, we developed a disulfiram-loaded emulsion to deliver the highly liposoluble drug. Treatment with the emulsion significantly delayed neuroblastoma progression in mice. These results identify KAT2A as a novel target of disulfiram, which directly impacts neuroblastoma epigenetics and is a promising candidate for repurposing to treat pediatric neuroblastoma.

Oyster protein hydrolysates alleviated chronic alcohol-induced liver injury in mice by regulating hepatic lipid metabolism and inflammation response

The oyster protein hydrolysate (OPH) possesses various biological activities that have the potential to ameliorate alcoholic liver disease (ALD). This study aimed to assess the protective effects of OPH on liver injury in mice induced by chronic alcohol treatment and the underlying mechanism was further explored by transcriptome and proteome from a global view. Compared with the Model group, OPH treatment significantly decreased the liver weight (p < 0.01) and reduced the content of liver injury markers alanine aminotransferase (ALT, by 34.14%, p < 0.01), aspartate aminotransferase (AST, by 35.31%, p < 0.01) and alkaline phosphatase (ALP, by 17.18%, p < 0.05) while increased the content of hepatic function marker total protein (TP, by 17.30%, p < 0.05) in serum. Meanwhile, only mild hepatocyte injury accompanied by less lipid droplet accumulation was observed in OPH treated ALD mice. The transcriptome and proteome results indicated that 482 target genes and 111 target proteins were involved in the ameliorative effect of OPH on ALD. After data integration, 43 co-regulated targets were identified, which were mainly related to lipid metabolism (reduction of cholesterol and triglyceride accumulation) and inflammatory response [inhibition of inflammatory responses through toll-like receptors (TLRs), nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs), and tumor necrosis factor (TNF) signaling pathways]. Consistent with omics data, the hepatic levels of total lipid, total cholesterol, triglyceride, interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), and transforming growth factor β (TGF-β) were declined by OPH treatment in ALD mice. Collectively, our results prove that OPH possesses potent hepatoprotective activities and has the potential to be used as a novel functional ingredient for the management of ALD.

Efficiency of the original flavonoids composition based on Curcumin in chronic alcohol intoxication

IntroductionMultiple tissue and organ damage induced by ethanol toxic effects requires prescription of a wide range of drugs that have a positive symptomatic effect, while simultaneously increasing the toxic load on the body, which necessitates the search for new approaches to the therapy of alcoholism, possibly with the use of parapharmaceuticals.ObjectivesThe anti-inflammatory, antioxidant and neuroprotective properties of curcuminoids, as well as their ability to influence the epigenetic mechanisms of gene expression regulation, can provide a positive therapeutic effect in alcoholism by influencing the pathogenetic mechanisms of this pathology, which determines the relevance and prospects of studying their effects during chronic ethanol intoxication.Methods(CBAxC57Bl/6) F1 male mice with 6-month 10% ethanol exposure were undergoing administration of original composition of flavonoids based on Curcumin for 40 days. Animal’s alcohol consumption, behavior and immune parameters were estimated.Results As a result of flavonoids original composition administration, correction of the changes in the nervous system functional activity, caused by the chronic toxic effect of ethanol has been achieved (decrease in alcoholic motivation, exploratory behavior stimulation, modulation of the cytokines level in brain, indicating a neuroinflammation decrease). We also registered the immune system functional activity modulation (stimulation of immune response and lymphocytes proliferative activity) in mice with alcohol dependence.ConclusionsThe original composition of flavonoids against the background of long-term alcohol consumption has a positive psychoneuroimmunomodulatory effect, which serves as an experimental substantiation of the prospects of using the composition as an adjuvant in the treatment of chronic alcoholism.DisclosureNo significant relationships.

Modern approaches in the treatment of chronic alcoholism

The article presents an analysis of information from literary sources devoted to the study of modern approaches in the treatment of chronic alcoholism. Chronic alcohol intoxication is characterized by a high degree of complications from the cardiovascular and nervous systems, as well as the liver and gastrointestinal tract. In this case, drugs are used mainly to relieve withdrawal symptoms. It is for the relief of withdrawal symptoms that pharmacotherapy is most often used. Since alcoholism causes psychiatric disorders that are extremely varied in their clinical manifestations and severity, a wide range of psychotropic drugs is used to treat them. In addition to this group, energy-correcting agents are also used for antihypoxic and detoxifying therapy. An indication for the use of drugs from the class of antioxidants, combining the properties of tranquilizers and nootropics, without exerting a toxic effect on the heart and protecting the myocardium under conditions of hypoxia, is alcohol withdrawal syndrome with a predominance of neurosis-like and vegetative-vascular disorders. However, at the moment there is a lack of information about the effectiveness of their use in alcohol withdrawal

Plate-like Alginate Microparticles with Disulfiram-SPIO-Coencapsulation: An In Vivo Study for Combined Therapy on Ovarian Cancer.

Disulfiram is a drug used to support the treatment of chronic alcoholism. Recently, it has been found to have an off-label ability to inhibit the growth of ovarian cancer cells. However, the original formulation was designed for use via oral administration, which is not suitable to be given by a direct spray on the affected area. Therefore, in this study, we designed and prepared alginate (ALG) microparticles loaded with disulfiram and superparamagnetic iron oxide (cross-linking disulfiram/SPIO/ALG MPs), which have great potential application for inhibiting the growth of ovarian cancer simultaneously via two treatments, i.e., chemotherapy and hyperthermia. The drug-encapsulating alginate microparticles were prepared using an electrospray system and then cross-linked with calcium chloride ions. The particles were observed by optical microscopy and scanning electron microscopy, and found to be approximately 200 μm in diameter. The disc-shape morphology of the microparticles could be controlled by up to 95%. The drug-encapsulation efficiency of the microparticles reached 98%, and the suppression of tumor growth for the free-form disulfiram-treated group and disulfiram/SPIO/ALG MPs-treated group were 48.2% and 55.9% of tumor volume reduction, respectively, compared with a cisplatin-treated group. A hyperthermic effect can be achieved by applying a magnetic field to oscillate SPIO. The results of this study showed that these cross-linking disulfiram/SPIO/ALG MPs are potential drug carriers for the treatment of ovarian cancer.

Alcohol-abuse drug disulfiram targets pediatric glioma via MLL degradation

Pediatric gliomas comprise a broad range of brain tumors derived from glial cells. While high-grade gliomas are often resistant to therapy and associated with a poor outcome, children with low-grade gliomas face a better prognosis. However, the treatment of low-grade gliomas is often associated with severe long-term adverse effects. This shows that there is a strong need for improved treatment approaches. Here, we highlight the potential for repurposing disulfiram to treat pediatric gliomas. Disulfiram is a drug used to support the treatment of chronic alcoholism and was found to be effective against diverse cancer types in preclinical studies. Our results show that disulfiram efficiently kills pediatric glioma cell lines as well as patient-derived glioma stem cells. We propose a novel mechanism of action to explain disulfiram’s anti-oncogenic activities by providing evidence that disulfiram induces the degradation of the oncoprotein MLL. Our results further reveal that disulfiram treatment and MLL downregulation induce similar responses at the level of histone modifications and gene expression, further strengthening that MLL is a key target of the drug and explaining its anti-oncogenic properties.

Nanomedicine Enables Drug-Potency Activation with Tumor Sensitivity and Hyperthermia Synergy in the Second Near-Infrared Biowindow

Disulfiram (DSF), a U.S. Food and Drug Administration (FDA)-approved drug for the treatment of chronic alcoholism, is also used as an antitumor drug in combination with Cu2+ ions. However, studies have shown that the endogenous Cu2+ dose in tumor tissues is still insufficient to form relatively high levels of a bis(N,N-diethyldithiocarbamate) copper(II) complex (denoted as Cu(DTC)2) to selectively eradicate cancer cells. Here, DSF-loaded hollow copper sulfide nanoparticles (DSF@PEG-HCuSNPs) were designed to achieve tumor microenvironment (TME)-activated in situ formation of cytotoxic Cu(DTC)2 for NIR-II-induced, photonic hyperthermia-enhanced, and DSF-initiated cancer chemotherapy. The acidic TME triggered the gradual degradation of DSF@PEG-HCuSNPs, promoting the rapid release of DSF and Cu2+ ions, causing the in situ formation of cytotoxic Cu(DTC)2, to achieve efficient DSF-based chemotherapy. Additionally, DSF@PEG-HCuSNPs exhibited a notably high photothermal conversion efficiency of 23.8% at the second near-infrared (NIR-II) biowindow, thus significantly inducing photonic hyperthermia to eliminate cancer cells. Both in vitro and in vivo studies confirmed the effective photonic hyperthermia-induced chemotherapeutic efficacy of DSF by integrating the in situ formation of toxic Cu(DTC)2 complexes and evident temperature elevation upon NIR-II laser irradiation. Thus, this study represents a distinctive paradigm of in situ Cu2+ chelation-initiated "nontoxicity-to-toxicity" transformation for photonic hyperthermia-augmented DSF-based cancer chemotherapy.

Possible ameliorative role of green tea on chronic alcohol mediated renal toxicity of STZ -induced diabetic rats

Summary This research was planned to assess the protective effect of aqueous green tea extract (AGTE) in alcohol treated diabetic rats. This study was examined, whether 20% v/v of 5 g/kg.b.w alcohol intake exacerbates diabetes-induced renal damage associated with an escalated nitroxidative stress and glucose homeostasis in comparison with diabetic condition alone. The present study is primarily focused to investigate the possible toxic effects of chronic alcohol intake on kidney of streptozotocin (STZ)-induced diabetic rats. Forty albino male Wistar rats were used and randomly allocated into five groups. Diabetes was induced by a single intraperitoneal injection of STZ (50 mg/kg) in rats. Alcohol was administered through orogastric tube once daily for a period of 60 days. Impact of chronic alcohol treatment on renal function and selected metabolites of catecholamines and antioxidant metabolisms have been investigated in the kidney of STZ-induced diabetic rats. Biochemical analysis of kidneys demonstrated that chronic alcohol induced alterations in vitro gluconeogenesis, significant decrease in the body weight, and activities of antioxidant enzymes in diabetic rats. A significant increase in the levels of epinephrine and sertonine was observed between alcohol treated, diabetic control and alcohol treated diabetic rat groups. Further, excessive chronic alcohol treatment caused additive deterioration of the kidney function as evidenced by increased inducible nitric oxide synthase (iNOS) protein expression and thus nitroxidative stress and histopathological aberrations leading to aggravation of diabetic nephropathy under alcohol intoxication. Treatment with aqueous green tea extract (300 mg/kg.b.w) to diabetic rats treated with alcohol renders the protection against the decreased body weight, increased hyperglycemia, hyperlipidemia, catecholamines, nitroxidative stress and further tissue damage.

The hepato-protective effect of eupatilin on an alcoholic liver disease model of rats.

ABSTRACT Eupatilin is known to possess anti-apoptotic, anti-oxidative, and anti-inflammatory properties. We report here that eupatilin has a protective effect on the ethanol-induced injury in rats. Sprague–Dawley rats were divided into 6 groups: control, vehicle, silymarin, eupatilin 10 mg/kg, eupatilin 30 mg/kg, and eupatilin 100 mg/kg. Plasma levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were analyzed to determine the extent of liver damage. Total cholesterol (TC) and triglycerides (TG) were analyzed to determine the level of liver steatosis. Malondialdehyde level, superoxide dismutase (SOD) activity, and glutathione (GSH) level were analyzed to determine the extent of oxidative stress. Tumor necrosis factor (TNF)-α and interleukin (IL)-1β were quantified to verify the degree of inflammation. Based on our findings, chronic alcohol treatment significantly changed the serum indexes and liver indicators of the model rats, which were significantly improved by eupatilin treatment. Rats in the eupatilin-treatment group showed reduced levels of AST, ALT, TG, TC, TNF-α, and IL-1β, increased SOD activity and GSH levels, and improved overall physiology compared to the alcoholic liver disease model rats. H&E staining also verified the eupatilin-mediated improvement in liver injury. In conclusion, eupatilin inhibits alcohol-induced liver injury via its antioxidant and anti-inflammatory effects.

Drugs Repurposing in High-Grade Serous Ovarian Cancer.

Ovary Carcinoma (OC) is the most lethal gynecological neoplasm due to the late diagnoses and to the common development of resistance to platinum-based chemotherapy. Thus, novel therapeutic approaches are urgently required. In this regard, the strategy of drug repurposing is becoming attractive. By this approach, the effectiveness of a drug originally developed for another indication is tested in a different pathology. The advantage is that data about pharmacokinetic properties and toxicity are already available. Thus, in principle, it is possible to reduce research costs and to speed up drug usage/marketing. Here, some noticeable examples of repurposed drugs for OC, such as amiodarone, ruxolitinib, statins, disulfiram, ormeloxifenem, and Quinacrine, are reported. Amiodarone, an antiarrhythmic agent, has shown promising anti-OC activity, although the systemic toxicity should not be neglected. The JAK inhibitor, Ruxolitinib, may be employed particularly in coadministration with standard OC therapy as it synergistically interacts with platinum-based drugs. Particularly interesting is the use of statin which represent one of the most commonly administered drugs in aged population to treat hypercholesterolemia. Disulfiram, employed in the treatment of chronic alcoholism, has shown anti-OC properties. Ormeloxifene, commonly used for contraception, seems to be promising, especially due to the negligible side effects. Finally, Quinacrine used as an antimicrobial and anti-inflammatory drug, is able to downregulate OC cell growth and promote cell death. Whereas further testing in patients are necessary to better clarify the therapeutic potential of repurposed drugs for OC, it is believed that their use, better if combined with OC targeted delivery systems, can significantly contribute to the development of novel and effective anti-OC treatments.

Cross talk between alcohol-induced oxidative stress and HCV replication.

Alcohol consumption exacerbates the pathogenesis of hepatitis C virus (HCV) infection and aggravates disease consequences in alcohol-abusing patients. Although the exact reasons by which alcohol consumption affects several cellular pathways in liver cells are not clear, they might be partially attributed to the ability of alcohol to further suppress the innate immunity, modulation of autophagy and also its relationship with reactive oxygen species (ROS) generation. To evaluate these issues, Huh7 cells harboring HCV replicon and Cytochrome p450 (CYP2E1) plasmid were exposed to ethanol and mRNA expression of Beclin-1, interferon-stimulated gene15 (ISG15) genesand HCV NS5B for two different times were relatively quantitated. ROS was determined by flow cytometry. The results showed that alcohol treatment in a short time caused an increase in HCV NS5B and Beclin-1 mRNA and decreased ISG 15 mRNA. Long-lasting alcohol treatment increased ROS production in Huh-7 cells and HCV replication was reduced. In conclusion, acute alcohol treatment might contribute to increase HCV replication by interference in innate immunity and induction of autophagy. Chronic alcohol treatment caused oxidative stress, which disrupts autophagy and thereby increased the rate of Huh7 cell injury.

Fluoxetine improves behavioural deficits induced by chronic alcohol treatment by alleviating RNA editing of 5-HT2C receptors

The alcoholism and major depressive disorder are common comorbidity, with alcohol-induced depressive symptoms being eased by selective serotonin re-uptake inhibitors (SSRIs), although the mechanisms underlying pathology and therapy are poorly understood. Chronic alcohol consumption affects the activity of serotonin 2C receptors (5-HT2CR) by regulating adenosine deaminases acting on RNA (ADARs) in neurons. Astrogliopathic changes contribute to alcohol addiction, while decreased release of ATP from astrocytes can trigger depressive-like behaviours in mice. In this study, we discovered that chronic alcohol treatment increased editing of RNA of 5-HT2CR via up-regulating the expression of ADAR2, consequently reducing the release of ATP from astrocytes induced by 5-HT2CR agonist, MK212. Moreover, SSRI antidepressant fluoxetine decreased the expression of ADAR2 through the transactivation of EGFR/PI3K/AKT/cFos signalling pathway. The increased release of astroglial ATP by MK212 which was suppressed by chronic alcohol consumption, and reduction in ADAR2 activity eliminated the RNA editing of 5-HT2CR increased by alcohol in vitro and recovered the release of ATP from astrocytes induced by MK212. Meanwhile, fluoxetine improved the behavioural and motor symptoms induced by alcohol addiction and decreased the alcohol intake. Our study suggests that the astrocytic 5-HT2CR contribute to alcohol addiction; fluoxetine thus can be used to alleviate depression, treat alcohol addiction and improve motor coordination.

Noble metal sensitized invasive porous bioelectrodes: advanced medical device for enhanced neuronal activity and chronic alcohol treatment.

Invasive bioelectrodes are widely used as an effective treatment for several acute and chronic diseases. In earlier work using high surface area invasive porous bioelectrodes evaluated in an animal model of alcoholism withdrawal, we demonstrated significantly improved electrophysiological and behavioral responses. In this study, we further modify the surface of these invasive porous bioelectrodes with noble metal (Ag, Au, Pt) nanoparticles. Compared to both conventional and porous bioelectrodes, noble metal sensitized invasive porous bioelectrodes show markedly increased low threshold (LT) and wide dynamic range (WDR) neuronal activity. In particular, Pt-sensitized invasive porous bioelectrodes show the highest WDR neuronal activity only upon insertion. In addition, Ag-sensitized invasive porous bioelectrodes, whose surface area is about 37 times greater than that of conventional bioelectrodes, show improved electrochemical properties with higher LT and WDR neuronal activity when stimulated. In an animal model of chronic alcoholism, using normal and alcohol-treated Sprague-Dawley (SD) rats evaluated with the elevated plus maze (EPM) test, the Ag-sensitized invasive porous bioelectrodes show about 20% higher open arms time. These results suggest that these noble metal-sensitized invasive bioelectrodes may offer improved therapeutic outcomes for the treatment of chronic alcoholism, and given these enhanced electrophysiological properties, for other conditions as well.

Detection of Human Pharmaceuticals in the Surface Water of East Fork Stones River

Pharmaceuticals have been detected nationwide in different environmental matrices including wastewater effluents and surface water. In recent studies, pharmaceuticals have also been found in aquatic plants, fish tissues and plasma of shark bulls. Pharmaceuticals that were detected as indicated in published literature, included steroids, prescription drugs such as antibiotics, anti-depressants, anti-inflammatory drugs, hormones and over the counter (OTC) drugs. We conducted a monitoring study to detect the presence of pharmaceuticals in East fork of the Stones River located in Rutherford County in Middle Tennessee. East Fork Stones River is a six (6) order river that drains a large portion of Rutherford County including Murfreesboro, Tennessee. Grab water samples were collected from Stones River for three seasons: (summer and fall of 2014 and 2015 and winter of 2015 and 2016) each year. Water quality parameters were also determined in situ using Eureka Water ProbesTM multi-parameter sondes. Water samples were analyzed for the presence of pharmaceutical compounds using GC-MS. Chemical Abstract Service Registry Numbers (CASRN or CAS) for detected pharmaceuticals were identified. Pharmaceuticals detected included those used for treatment of chronic alcoholism (Disulfiram: CAS # 97-77-8), a compound (Thiazolidine: CAS # 504-78-9) in the drug Thiazolidinedione which is used for the treatment of type II diabetes, a compound associated with the prevention of anti-inflammatory conditions (Methyl palmitate: CAS # 112-39-0), and emollient in skincare (Undecane: CAS # 1120-21-4). While the quantitative concentrations of these drugs were not determined in this study, their qualitative presence in surface water is noteworthy.

Ethanol induced increase of fibroblast growth factor 2 mRNA content in emotiogenic brain structures of rats

We studied the effects of acute, subacute, and chronic alcohol treatment of rats on the content of fibroblast growth factor 2 (FGF2) mRNA in various brain structures. Results suggest a possible role of FGF2 in the functioning of dopaminergic neurons in the midbrain. In our experiment, ethanol treatment of rats was accompanied by an increase in the FGF2 mRNA level in the emotiogenic structures of the brain. This effect was blocked by pretreatment of animals with chlorpromazine. This suggests FGF2 involvement in the mechanisms of alcohol dependence and can be considered as a possible diagnostic and therapeutic target in alcoholism.

Pharmacotherapeutic aspects of nootropics use in people with alcohol dependence

In the pharmacotherapy of chronic alcoholism a significant place is given to nootropic drugs. Currently, neurodegenerative processes are associated with the activity of various neurotrophic factors of the brain and neuropeptides. An important advantage of the use of neuropeptide drugs (cerebrolysin, semax, cortexin) in the treatment of chronic alcoholism is that they have an organ-specific multimodal effect on the brain: provide metabolic regulation, neuroprotection, functional neuromodulation and neurotrophic activity. They are recommended for relief of acute withdrawal syndrome, as well as in the post-abstinent period in asthenic syndrome and restoration of mnestic functions.

PDTM-06. DISULFIRAM-COPPER PROLONGS SURVIVAL AND REDUCES STEM-CELLNESS IN PEDIATRIC GROUP 3 MEDULLOBLASTOMA THROUGH NUCLEAR SEQUESTRATION OF NPL4

Abstract Medulloblastoma (MB), an aggressive pediatric brain malignancy, is commonly classified into four subtypes. Although survival has improved remarkably, Group 3 and Shh-driven tumors still experience high rates of recurrent and metastatic disease. Disulfiram (DSF), a safe and inexpensive drug, has shown an antitumor effect when administered with copper (Cu++) against other brain malignancies, and its repurposing to MB could benefit current standards of treatment. ONS76 (Shh-driven Tp53 wild-type), UW228 (Shh-driven Tp53-mutated), D425med and D341med (Group 3) lines were treated with DSF-Cu++and tested with viability, proliferation and clonogenic assays. Flow cytometry (FACS) with AnnexinV-PI, Ki67, PI/RNAse for necrosis and cell cycle, and CD133 and Aldefluor for cell-stemness were carried out. MB lines were tested with western blotting (WB) and immunofluorescence (IF) for NPL4(Nuclear protein localization protein 4 homolog), AIF (apoptosis-inducing factor), cleaved-PARP(Poly-ADP-ribose-polymerase), H2AX and other molecular targets. In vivo safety (n=8) and efficacy studies (n=16) were carried out on Nu/Nu athymic mice implanted intracranially (IC) with D425med cells, along with Hematoxylin/Eosin and immunohistochemistry analyses on brain samples. DSF-Cu++showed remarkable cytotoxic and anti-proliferative activity against all MB lines. In particular, NPL4, a protein involved in the ubiquitin signaling, appears to be the primary DSF target, as previously shown in other malignancies. Apoptosis quickly follows the Heat-Shock Response, as confirmed by FACS, WB and IF, and cell-stemness is decreased significantly by nanomolar doses of DSF-Cu++(p=0.002). Oral administration of DSF-Cu++was proven safe at doses of 100 mg/kg and significantly prolonged survival in IC xenografts (median 28.5 days; p=0.02) compared to controls (median 24 days). DSF, a compound used in the treatment of chronic alcoholism, has recently shown a remarkable effect in a number of malignancies when co-administered with Cu++. We provide proof of its potential as a chemotherapeutic for pediatric MB, and describe its molecular mechanism of action.