Effects of alcohol mixed with energy drink and alcohol alone on subjective intoxication

Recent studies suggest that the combination of caffeine-containing drinks together with alcohol might reduce the subjective feelings of alcohol intoxication—the so-called “masking effect”. In this study, we aimed to review the effects of alcohol in combination with caffeine or energy drink with special focus on the “masking effect”. Fifty-two healthy male volunteers were analysed concerning breath alcohol concentration and subjective sensations of intoxication using a 18 item Visual Analogue Scale in a randomised, double-blinded, controlled, four treatments cross-over trial after consumption of (A) placebo, (B) alcohol (vodka 37.5 % at a dose of 46.5 g ethanol), (C) alcohol in combination with caffeine at a dose of 80 mg (equivalent to one 250 ml can of energy drink) and (D) alcohol in combination with energy drink at a dose of 250 ml (one can). Primary variables were headache, weakness, salivation and motor coordination. Out of four primary variables, weakness and motor coordination showed a statistically significant difference between alcohol and non-alcohol group, out of 14 secondary variables, five more variables (dizziness, alterations in sight, alterations in walking, agitation and alterations in speech) also showed significant differences due mainly to contrasts with the non-alcohol group. In none of these end points, could a statistically significant effect be found for the additional ingestion of energy drink or caffeine on the subjective feelings of alcohol intoxication. This within-subjects study does not confirm the presence of a “masking effect” when combining caffeine or energy drink with alcohol.     

Alcohol-induced liver cirrhosis as a cause of portal hypertension

Basic data on pathomorphology and symptomatology of the alcohol-induced liver cirrhosis accompanied by portal hypertension are discussed. Respective data were compared with the group of cirrhotic patients not abusing alcohol. A high percentage of encephalopathic disorders and nearly 50% of the patients suffering from the hemorrhage from esophageal varices were the first sign of the cirrhosis in both groups. Despite hemorrhage from esophageal varices a few patients obtained surgical help preventing recurrence of the hemorrhage. Liver functional reserve, incidence of encephalopathies and the degree of liver involvement are in favour for non-alcohol cirrhosis. Inflammatory process in the liver, splenomegaly and hypersplenism were more frequent in the liver cirrhosis of non-alcohol origin.     

Isotopic probes into pathways of ethanol metabolism

The relative extent of tritium labeling in glucose and water was determined when l-[2-³H]lactate or [(1R)1-³H]ethanol were the labeled substrates for rat liver parenchymal cells, incubated with 20 mm ethanol and 10 mml-lactate. From the relatively lower specific yield in glucose from the tritiated ethanol one can calculate a percentage contribution of a non-alcohol dehydrogenase-mediated pathway to total ethanol metabolism. This calculated value (about 35%) is somewhat higher than that determined by the use of pyrazole at 5 mm to inhibit alcohol dehydrogenase. The utilization of [(1R)1-³H]ethanol is slower than that of unlabeled ethanol, both in the absence and presence of 5 mm pyrazole, indicating isotope discrimination against tritium in both the alcohol dehydrogenase and non-alcohol dehydrogenase pathways.There was only a slight difference in the rate of utilization of normal and fully deuterated ethanol by rat liver cells in the absence of pyrazole. However, in the presence of 5 mm pyrazole, where essentially only the non-alcohol dehydrogenase pathway operates, deuterated ethanol was utilized at only about half the rate of nondeuterated ethanol. These findings are difficult to reconcile with a catalase-mediated pathway of ethanol metabolism in which the rate-limiting factor is the rate of H₂O₂ generation.     

Effect of chronic alcohol consumption on rat brain microsome lipid composition, membrane fluidity and Na+-K+-ATPase activity

The present study reports differences in phospholipid classes, fatty acids of individual phospholipids, and changes in membrane fluidity and Na+-K+-ATPase activity in brain microsomes of rats maintained on an alcohol diet for 35 days compared to sex, age and weight-matched control rats maintained on a calorically-equivalent, non-alcohol diet. Although no difference in Na+-K+-ATPase activity was found in microsomes from alcohol vs control rats when measured in the absence of added alcohol, the presence of low concentrations of ethanol (less than 100 mM) stimulated, while high concentrations (greater than 100 mM) inhibited enzyme activity. The stimulation was differentially expressed in that the microsomal enzyme from alcohol rats was stimulated to a lesser extent than the enzyme from control rats. However, the inhibiting effect of high concentrations of alcohol was similar in microsomes from both alcohol and control rats. Also in membranes from alcohol rats, there was a lower quantity of phosphatidylethanolamine (PE) and higher quantities of phosphatidylserine (PS) and phosphatidylinositol (PI) compared to membranes from control rats. The major change in fatty acid composition was a reduction in the level of polyunsaturated fatty acids, which was particularly evident in PI and PS. The linoleic acid: arachidonic acid ratio (18:2/20:4) and the saturation:unsaturation ratio were also increased in PI and PS in membranes from alcohol animals. However, the ratio of n-6/n-3 fatty acids remained the same or was reduced in membranes from alcoholic animals. Although no difference in the inherent "fluidity" of membranes from alcohol vs control rats could be demonstrated by electron paramagnetic resonance, molecular tolerance to ethanol was demonstrated in the membranes from alcohol rats by the resistance to the disordering effects of added ethanol.     

Synthesis and degradation of alcohol dehydrogenase in wild-type and Adh-null activity mutants of Drosophila melanogaster

Both the amount and the size of alcohol dehydrogenase-like cross-reacting material was determined in 14 ethyl methanesulfonate (EMS)-induced alcohol dehydrogenase-null activity mutants. In 11 mutants cross-reacting material of the same apparent molecular weight as alcohol dehydrogenase was detected, while in 3 mutants no cross-reacting material was found. In all cases, the amount of cross-reacting material found in the mutants was lower than that in wild-type flies. High, intermediate, and low cross-reacting material-producing mutants showed similar initial rates of incorporation of labeled amino acid into alcohol dehydrogenase-like protein, presumably reflecting similar rates of synthesis. If the rate of synthesis of cross-reacting material is the same in the mutants as in the wild type, then the different levels of cross-reacting material must be due to different rates of degradation.Supported by NIH Grants GM-18254 and ES-01527 and DOE Contract EY-76-S-2965.     

Alcohol teratogenesis: mechanisms of damage and strategies for intervention

There are multiple mechanisms by which alcohol can damage the developing brain, but the type of damage induced will depend on the amount and developmental timing of exposure, along with other maternal and genetic factors. This article reviews current perspectives on how ethanol can produce neuroteratogenic effects by its interactions with molecular regulators of brain development. The current evidence suggests that alcohol produces many of its damaging effects by exerting specific actions on molecules that regulate key developmental processes (e.g., L1 cell adhesion molecule, alcohol dehydrogenase, catalase), interfering with the early development of midline serotonergic neurons and disrupting their regulatory-signaling function for other target brain structures, interfering with trophic factors that regulate neurogenesis and cell survival, or inducing excessive cell death via oxidative stress or activation of caspase-3 proteases. The current understanding of pathogenesis mechanisms suggests several strategic approaches to develop rational molecular prevention. However, the development of behavioral and biologic treatments for alcohol-affected children is crucial because it is unlikely that effective delivery of preventative interventions can realistically be achieved in ways to prevent prenatal damage in at-risk pregnancies. Toward that end, behavioral training that promotes experience-dependent neuroplasticity has been effective in a rat model of cerebellar damage induced by alcohol exposure during the period of brain development that is comparable to that of the human third trimester.     

Oxidation product(s) in acetaldehyde reacts with NAD(P)H and interferes with assay of alcohol dehydrogenase

A decrease in absorbance at 340 nm, at rates similar to those obtained with alcohol dehydrogenases in routine assays, occurred when NADH or NADPH was mixed with acetaldehyde that had been exposed to air for various durations. NAD(P)H was apparently oxidized by interfering substance(s) present in acetaldehyde. Reagent-grade acetaldehyde from newly opened bottles as well as acetaldehyde redistilled under strictly O2-free conditions contained minimal amounts of NAD(P)H-reacting substance(s). Redistillation under poor anaerobic conditions or in air increased the amount of NAD(P)H-reacting substance(s) in redistilled acetaldehyde. NADPH reacted at a higher rate than NADH with the interfering substance(s) in Tris-Cl buffer at pH 7.5. Also, the reaction was faster in Tris buffer than in phosphate buffer at pH 7.5. The NAD(P)H-oxidizing reaction may not be apparent when the nominal concentration of acetaldehyde used was below 5 mM, but the measured ethanol dehydrogenase activity could be significantly lower with acetaldehyde containing a measurable level of interfering substance(s). This study suggests that acetaldehyde is most easily tested with NADPH for the presence of a significant level of interfering substance(s) and that redistillation, if necessary, must be performed under strictly O2-free conditions.     

Beer and health: preventive effects of beer components on lifestyle-related diseases

It has been demonstrated that the light-to-moderate consumption of alcoholic beverages is associated with significant reductions in all-cause and particularly cardiovascular mortality. While the inverse association between red-wine consumption and cardiovascular risk is globally recognized as the French paradox, many epidemiological studies have concluded that beer and red wine are equally beneficial. Moderate alcohol intake improves lipoprotein metabolism and lowers cardiovascular mortality risk. The question now is whether additional health benefits associated with the non-alcohol components in beer may be expected. This article summarizes the results of the latest studies on the health benefits of beer while referring to our recent results, which demonstrate the preventive effects of beer and its components on lifestyle-related diseases. A series of studies using animal models have shown that beer may prevent carcinogenesis and osteoporosis; beer provides plasma with significant protection from oxidative stress; and isohumulones, the bitter substances derived from hops, may prevent and improve obesity and type-2 diabetes, improve lipid metabolism, and suppress atherosclerosis. Further studies are needed to clarify the components in addition to isohumulones that are responsible for these beneficial effects of beer, and the underlying mechanisms must be addressed.     

Rapid Dehydration for Celloidin Embedding by Means of Azeotropic Distiliation

By including plant material in a fractional distillation apparatus containing 95% alcohol and benzene it is possible to eliminate water completely in a constant boiling mixture. The dehydrated material can then be embedded in celloidin by orthodox techniques. In spite of the rapidity of the dehydration quite soft tissue as well as woody material can be processed. The method is cheap because it does not require absolute alcohol and is also suited to the humid climate in which it was developed.     

A Method for Injecting Insect Tracheae Permanently

By the use of an alcohol insoluble dye (trypan blue), acetic acid, and a detergent (“Santomerse No. 3”), a resulting dye solution is obtained which will completely penetrate the tracheal system of an insect. The dye is injected by the use of a vacuum and by the pressure produced when the air is allowed to re-enter the dye vessel. The dye itself is permanently fixed in the tracheae by means of a fixing solution containing alcohol, acetic acid and barium chloride as its components. The material must be properly preserved after staining. It may be stored indefinitely in 70% alcohol, xylol, cedar oil or clove oil, depending on whether the material is to be used for sectioning or for whole mounts. Injected material may be sectioned in either celloidin or paraffin, or may be cleared and mounted in toto.     

A Technic for Differential Staining of Nucleoli and Chromosomes

A procedure is described which enables a stain to be definitely located in the substance of the nucleolus. Material is fixed in either Navashin or Levitsky; the chromatin is stained by means of the improved Feulgen technic introduced by de Tomasi, and preparations brought thru the washing solutions down to distilled water. From distilled water the material is transferred to a mordant solution, 5% sodium carbonate in water, in which it is left for at least one hour. After mordanting wash well with water then stain for ten minutes in light green solution (90% alcohol, 100 cc, light green SFY, 0.5 g, aniline oil, 2 drops, well shaken); differentiate in alcoholic sodium carbonate solution, (70% alcohol saturated with carbonate); treat with 95% alcohol, absolute alcohol, equal parts xylene and absolute alcohol, clear in pure dry xylene and mount in neutral balsam. Cytoplasm and karyolymph should be quite clear, with magenta chromatin and well defined green nucleoli. The light green does not behave like a simple counterstain as in previous technics but as a definite stain for nucleolar material.     

Removal of Yolk from Oyster Eggs by Soxhlet Extraction for Clear Chromosome Preparations

The mature, spawned eggs of the American Eastern oyster, Crassostrea virginica Gmelin, contain yolk which interferes with the preparation of chromosomes and nuclear groups for the study of meiosis, fertilization, and karyology. Most samples of eggs cannot be studied cytogenetically until the yolk is extracted. Simple and complete removal of all interfering yolky material can be efficiently accomplished, after Carnoy fixation in 3:1 alcohol-acetic acid, by extraction for 2 hr in a micro-Soxhlet apparatus with 1:1 mixture of chloroform and methyl alcohol. Standard orcein squashes can then be routinely made of these eggs hitherto considered refractory to clear staining of chromosomes. A thimble with a fritted glass end, pore size 40 μ, is used as a receptacle for the eggs in the Soxhlet apparatus. After extraction the eggs can readily be washed off the fritted glass with the aceto-orcein staining solution. If the eggs are to be stored for some time prior to squashing and staining, 45-60% acetic acid or Carnoy's alcohol-acetic acid 3:1 can be used for the storage fluid.     

Ethanol induces oxidative stress in alveolar macrophages via upregulation of NADPH oxidases

Chronic alcohol abuse is a comorbid variable of acute respiratory distress syndrome. Previous studies showed that, in the lung, chronic alcohol consumption increased oxidative stress and impaired alveolar macrophage (AM) function. NADPH oxidases (Noxes) are the main source of reactive oxygen species in AMs. Therefore, we hypothesized that chronic alcohol consumption increases AM oxidant stress through modulation of Nox1, Nox2, and Nox4 expression. AMs were isolated from male C57BL/6J mice, aged 8-10 wk, which were treated with or without ethanol in drinking water (20% w/v, 12 wk). MH-S cells, a mouse AM cell line, were treated with or without ethanol (0.08%, 3 d) for in vitro studies. Selected cells were treated with apocynin (300 μM), a Nox1 and Nox2 complex formation inhibitor, or were transfected with Nox small interfering RNAs (20-35 nM), before ethanol exposure. Human AMs were isolated from alcoholic and control patients' bronchoalveolar lavage fluid. Nox mRNA levels (quantitative RT-PCR), protein levels (Western blot and immunostaining), oxidative stress (2',7'-dichlorofluorescein-diacetate and Amplex Red analysis), and phagocytosis (Staphylococcus aureus internalization) were measured. Chronic alcohol increased Nox expression and oxidative stress in mouse AMs in vivo and in vitro. Experiments using apocynin and Nox small interfering RNAs demonstrated that ethanol-induced Nox4 expression, oxidative stress, and AM dysfunction were modulated through Nox1 and Nox2 upregulation. Further, Nox1, Nox2, and Nox4 protein levels were augmented in human AMs from alcoholic patients compared with control subjects. Ethanol induces AM oxidative stress initially through upregulation of Nox1 and Nox2 with downstream Nox4 upregulation and subsequent impairment of AM function.     

A Paraffin Method for Refractory Plant Materials

A modification of Zirkle's n-butyl alcohol method for dehydrating refractory plant material and embedding it in paraffin is described. The material is cut into small pieces and killed and fixed in CRAF III. It is dehydrated in Zirkle's n-butyl alcohol series, slightly modified. Infiltration is accomplished by adding chips of paraffin of low melting point (52°C.) to the bottle containing butyl alcohol and the tissues at a gradually increasing series of temperatures. The butyl-alcohol-paraffin mixture is gradually replaced by pure paraffin (melting point 56-58°C). The material is embedded in Fisher Tissuemat (56-58°C). Before microtoming, the block of embedded tissue is trimmed so that part of the specimen is exposed, and it is soaked in water until it cuts easily.     

Comparison of two assay procedures for lignin peroxidase

The most widely accepted assay for detecting lignin peroxidase, based on the oxidation of veratryl alcohol to veratraldehyde, suffers from some drawbacks. At 310 nm, the wavelength at which the assay is performed, some other materials like lignins, quinonic compounds and aromatics also exhibit strong absorbance thus interfering with the estimation when present in the media. The present study reports the lignin peroxidase production by some white rot fungi under different nutritional conditions. The veratryl alcohol oxidation assay procedure for lignin peroxidase has been compared with another method based on the oxidation of the dye azure B involving absorbance measurements in the visible range. The latter method proved to be much more advantageous over the veratryl alcohol oxidation method, in media supplemented with malt extract, lignin preparations and agricultural residues. The enzyme production by veratryl alcohol assay could be detected only in mineral salts broth. By the azure B assay the enzyme activity was detected in all the media tested. The supplements gave varied response in different media. Veratryl alcohol enhanced the enzyme production in malt extract broth and mineral salts malt extract broth. Among the lignin preparations Indulin AT increased the lignin peroxidase titres from 2 to 20 fold in different fungi. Similarly, wheat straw supplemented in mineral salts broth and malt extract broth, separately, strongly stimulated the lignin peroxidase production. The above studies revealed that azure B assay may act as a substitute or equivalent method.     

Solvent comparison in the isolation, solubilization, and toxicity of Stachybotrys chartarum spore trichothecene mycotoxins in an established in vitro luminescence protein translation inhibition assay

It is well known that non-viable mold contaminants such as macrocyclic trichothecene mycotoxins of Stachybotrys chartarum are highly toxinigenic to humans. However, the method of recovering native mycotoxin has been without consensus. Inconsistencies occur in the methods of isolation, suspension, preparation, and quantitation of the mycotoxin from the spores. The purpose of this study was to provide quantitatively comparative data on three concurrent preparations of 10(6)S. chartarum spores. The experiments were designed to specifically evaluate a novel method of mycotoxin extraction, solubilization, and the subsequent inhibitory effect in an established in vitro luminescence protein translation assay from 30 day-old spores. The mycotoxin-containing spores swabbed from wallboard cultures were milled with and without glass beads in 100% methanol, 95% ethanol, or water. Milled spore lysates were cleared of cell debris by filter centrifugation followed by a second centrifugation through a 5000 MWCO filter to remove interfering proteins and RNases. Cleared lysate was concentrated by centrivap and suspended in either alcohol or water as described. The suspensions were used immediately in the in vitro luminescence protein translation assay with the trichothecene, T-2 toxin, as a control. Although, mycotoxin is reported to be alcohol soluble, the level of translation inhibition was not reliably satisfactory for either the methanol or ethanol preparations. In fact, the methanol and ethanol control reactions were not significantly different than the alcohol prepared spore samples. In addition, we observed that increasing amounts of either alcohol inhibited the reaction in a dose dependent manner. This suggests that although alcohol isolation of mycotoxin is desirable in terms of time and labor, the presence of alcohol in the luminescence protein translation reaction was not acceptable. Conversely, water extraction of mycotoxin demonstrated a dose dependent response, and there was significant difference between the water controls and the water extracted mycotoxin reactions. In our hands, water was the best extraction agent for mycotoxin when using this specific luminescence protein translation assay kit.     

Labeling methods for the study of poly- and mono(ADP-ribose) metabolism in cultured cells

Methods are described for the radiolabeling and determination of NAD+, poly(ADP-ribose), and protein-bound monomers of ADP-ribose in cultured mammalian cells. The adenine nucleotide pools of confluent monolayer cell cultures are radiolabeled using high-specific-activity [3H]adenine. Following any desired experimental manipulation, cultures are treated with trichloroacetic acid. Radiolabel in NAD+ can be rapidly determined from the acid-soluble fraction using dihydroxyboronyl Sepharose (DHB-Sepharose). The acid-insoluble material can be analyzed for radiolabeled polymers of ADP-ribose and protein-bound monomers of ADP-ribose. Polymers are separated from interfering material using dihydroxyboronyl-Bio-Rex 70 (DHB-Bio-Rex). Protein-bound monomers are separated from noncovalently bound ADP-ribose and different classes of (ADP-ribosyl) protein linkages are released by specific chemical treatments. The released ADP-ribose is then separated from interfering materials using DHB-Bio-Rex and DHB-Sepharose. Control experiments have demonstrated the sensitivity, selectivity, and precision of the methods. Major advantages of the methods are that they allow many simultaneous determinations and all components can be determined from material derived from a single dish of cultured cells. The methods should prove useful for detailed studies of the metabolism of both protein-bound monomers and polymers of ADP-ribose in cultured mammalian cells.     

Chong木总皂甙的提取研究及毒理学实验

以秦岭五加科植物Chong木之茎皮为原料,用乙醇提取活性物质Chong木总皂甙,探讨了原料粒度大小,提取剂浓度,提取温度,提取时间与次数,料液比等对Chong木总皂甙提取率的影响,获得了最佳工艺条件;对Chong木总皂甙的急性毒性的毒理学试验表明,该提取物基本无毒,初步认为以其作为功能性食品基料资源开发,在一定程度上是安全的。     

Laboratory hints from the Literature: Department Devoted to Abstracts of books and papers from other Journals Dealing with stains and Microscopic Technic in General

A modification of Zirkle's n-butyl alcohol method for dehydrating refractory plant material and embedding it in paraffin is described. The material is cut into small pieces and killed and fixed in CRAF III. It is dehydrated in Zirkle's n-butyl alcohol series, slightly modified. Infiltration is accomplished by adding chips of paraffin of low melting point (52°C.) to the bottle containing butyl alcohol and the tissues at a gradually increasing series of temperatures. The butyl-alcohol-paraffin mixture is gradually replaced by pure paraffin (melting point 56-58°C). The material is embedded in Fisher Tissuemat (56-58°C). Before microtoming, the block of embedded tissue is trimmed so that part of the specimen is exposed, and it is soaked in water until it cuts easily.