• Energy Research

Those who work with cell cultures are certainly familiar-ized with the ritual of sanitizing everything that goesinto the cabinet or into the incubators with 70% ethanol,as is recommended by all the rules of aseptic techniquesand good cell culture practices. In spite of its eVective-ness in preventing microbiological contamination of thecultures, this gesture may have some undesirableconsequences.Probably, the research groups that study the eVects ofethanol on cell cultures are more conscious about therisks of sanitizing all the material with 70% ethanolbecause they need to guarantee that they have ethanol-free control samples. Thus, these groups routinely quan-tify ethanol in the culture medium to con Wrm the ethanolconcentrations used in their studies. However, the scien-tiWc community must be aware that when 70% ethanol isused on cell culture as a surface sanitization process, ifcautions are not taken, the cells will be exposed to highethanol concentrations. These high concentrations areachieved because, under normal cell culture conditions,the cell culture plates have appropriate venting systems(as they should have, otherwise the cells will not be oxy-genated) and generally, both cabinets and incubatorshave an ethanol-saturated atmosphere due to thefrequent sanitization procedures. Without additionalprecautions, ethanol concentrations of 0.5 mM are easilyattained, and depending on the ethanol-saturation of thecabinets or incubators, these concentrations can reachvalues of 5 mM as illustrated in Fig.1, which representsthe ethanol concentrations detected on 14 independentexperiments where no precautions were taken to avoidthe exposure of the medium to ethanol. Both theseconcentrations have proven eVects on cell function(Charness et al. 1994; Smith and Gong 2007). In fact, itis well described that ethanol, as a microsomal and mito-chondrial enzymatic inducer (CYP2E1, ADH-I), caninXuence the metabolism of several endogenous andexogenous compounds causing eventual antagonisms,synergisms or potentiations that will necessarily aVectthe Wnal results of the experiments (Jang and Harris2007).In addition, by itself, ethanol can directly or indirectlyalter the normal cell function, inXuencing cell survival,redox status, receptor functions, mitochondrial homeosta-sis, cell signaling, among others (Wu et al. 2006), and alsoaVecting the activity of infectious agents such as hepatitic Cvirus (Trujillo-Murillo et al. 2007).Thus, some precautions must be taken to minimize this“contamination” with ethanol: 70% ethanol used to sanitizesurface and objects should be completely dry before theXasks or plates are opened and these should not be sprayedwith 70% ethanol between coming out of the cabinet andgoing into the incubator (but obviously, in between, contactwith other surfaces is forbidden).These concerns are crucial for every lab working withcell culture and must be taken into account during the data

Quercus suber L. bark (cork) is a sustainable material due to its ability to regenerate. The aim of this work was to explore cork powders, by-products of the cork industry, as sustainable sources of value-added compounds. Two types of cork powder were studied: coarse (P0) and fine (P1). A broad physicochemical characterization was carried out, regarding particle size, color, moisture content, hygroscopicity, pH, heavy metal content, NIR spectra, and volatile compounds. DPPH scavenging activity and total phenolic content were also evaluated for an ethanolic P1 extract. For both powders, Hg, As, Cd and Pb contents were below the acceptable limits and volatile compounds commonly used as fragrances were found in their composition. P1 had a smaller and more homogenous particle size, lighter brownish color, lower pH value, and lower moisture content than P0, and therefore appears to be more suitable for industrial application. P1 ethanolic extract also showed a high scavenging activity and a content of phenolic compounds of 50.46 ± 0.63 mg (as gallic acid equivalents) per g of extract. In conclusion, P1 cork powder seems to be a promising source of upcycled ingredients, such as fragrances and antioxidants, for the pharmaceutical, nutraceutical, and cosmetic industries.

beta-Carotene and six xanthophylls (lutein, neoxanthin, violaxanthin, luteoxanthin, cryptoxanthin, and echinenone) have been identified and semiquantitatively or quantitatively determined in musts and port wines for the first time. An HPLC method was developed and compared with that of one based on thin layer cromatography with scanning densitometry. The most abundant carotenoids present in red grape varieties are beta-carotene and lutein. In wines, significant quantities of violaxanthin, luteoxanthin, and neoxanthin were found. This study was done with berries (skin and pulp), musts, and fortified wines. Some experiments were performed to follow carotenoid content from grapes to wines. Although the levels of beta-carotene and lutein found in fortified wines were lower than those found in musts, other xanthophylls, such as neoxanthin, violaxanthin, and luteoxanthin, exist in appreciable amounts in young ports.

It is becoming increasingly evident that ingested products, such as wine, may have profound effects on the therapeutic efficacy of certain drugs. As various xeno- and endobiotics are organic cations, the purpose of our study was to examine the modulation of organic cations intestinal apical uptake by red (RW) and white wine (WW). For this purpose, we used RW, WW, the same alcohol-free wines, phenolic compounds and ethanol. The uptake of the organic cation 1-methyl-4-phenylpyridinium (MPP+) was evaluated in Caco-2 cells, an intestinal epithelial cell model. RW and alcohol-free RW increased 3H-MPP+ apical uptake, although the effect of alcohol-free RW was less pronounced. On the other hand, WW and alcohol-free WW decreased the organic cation uptake but the effect of alcohol-free WW was more pronounced. Our results show that the total content in phenolic compounds was 7 times higher, and the dialysis index was about 4 times higher in RW compared to WW. Ethanol, in the same concentration found in wine, caused a significant decrease in 3H-MPP+ apical uptake. The solution containing high molecular weight compounds from dialyzed RW increased 3H-MPP+ apical uptake. In conclusion, the results suggest that RW may increase and WW may reduce the intestinal absorption of organic cations present in the diet, such as drugs or vitamins (e.g. thiamine and riboflavin). As ethanol alone decreased the uptake of MPP+, and alcohol-free RW and WW had a lower potency than intact wine upon the transport, the presence of ethanol is probably important for the solubilisation/bioavailability of the components endowed with the transport modulating activity.

3,4-Methylenedioxymethamphetamine (MDMA; ecstasy) is an amphetamine derivative drug with entactogenic, empathogenic and hallucinogenic properties, commonly consumed at rave parties in a polydrug abuse pattern, especially with cannabis, tobacco and ethanol. Since both MDMA and ethanol may cause deleterious effects to the liver, the evaluation of their putative hepatotoxic interaction is of great interest, especially considering that most of the MDMA users are regular ethanol consumers. Thus, the aim of the present study was to evaluate, in vivo, the acute hepatotoxic effects of MDMA (10mg/kg i.p.) in CD-1 mice previously exposed to 12% ethanol as drinking fluid (for 8 weeks). Body temperature was continuously measured for 12h after MDMA administration and, after 24h, hepatic damage was evaluated. The administration of MDMA to non pre-treated mice resulted in sustained hyperthermia, which was significantly increased in ethanol pre-exposed mice. A correspondent higher increase of hepatic heat shock transcription factor (HSF-1) activation was also observed in the latter group. Furthermore, MDMA administration resulted in liver damage as confirmed by histological analysis, slight decrease in liver weight and increased plasma transaminases levels. These hepatotoxic effects were also exacerbated when mice were pre-treated with ethanol. The activities of some antioxidant enzymes (such as SOD, GPx and Catalase) were modified by ethanol, MDMA and their joint action. The hepatotoxicity resulting from the simultaneous exposure to MDMA and ethanol was associated with a higher activation of NF-kappaB, indicating a pro-inflammatory effect in this organ. In conclusion, the obtained results strongly suggest that the consumption of ethanol increases the hyperthermic and hepatotoxic effects associated with MDMA abuse.

3,4-Methylenedioxymethamphetamine (MDMA; ecstasy), a drug of abuse commonly consumed at rave parties, is often taken in a polydrug abuse scenario, ethanol being one of the most associated drugs. Both MDMA and ethanol are mainly metabolized in the liver with formation of toxic metabolites. Our working hypothesis is that ethanol can modify the metabolism of MDMA through the cytochrome P450 system, and that this effect may be further potentiated by hyperthermia, a well-known consequence of MDMA abuse. To investigate these putative interactions we used primary rat hepatocyte cultures, which were exposed to 300 mM ethanol, 1.6 mM MDMA and the combination of both, at normothermic (36.5 degrees C) and hyperthermic (40.5 degrees C) conditions. After 24 h, the levels of MDA, HMA and HMMA in the cell culture medium were quantified by GC/MS. In addition, we repeated the same experimental design preceded by 1h incubation with 0.18 microM ketoconazole or 150 microM diallyl sulphide (CYP3A and CYP2E1 inhibitors, respectively), to evaluate the putative role of these isoenzymes in the observed effects. The results obtained showed that ethanol exposure increases the formation of some MDMA metabolites such as HMA (1.8 times increase) and MDA (1.5 times increase). This effect was markedly increased under hyperthermic conditions (HMA, MDA and HMMA formation increased 10, 6 and 16 times, respectively) and is mediated, at least partially, by CYP3A and CYP2E1.