Absence Of Ethanol Research Articles

An Optimized Method for the Measurement of Acetaldehyde by High‐Performance Liquid Chromatography

Acetaldehyde is produced during ethanol metabolism predominantly in the liver by alcohol dehydrogenase and rapidly eliminated by oxidation to acetate via aldehyde dehydrogenase. Assessment of circulating acetaldehyde levels in biological matrices is performed by headspace gas chromatography and reverse phase high-performance liquid chromatography (RP-HPLC). We have developed an optimized method for the measurement of acetaldehyde by RP-HPLC in hepatoma cell culture medium, blood, and plasma. After sample deproteinization, acetaldehyde was derivatized with 2,4-dinitrophenylhydrazine (DNPH). The reaction was optimized for pH, amount of derivatization reagent, time, and temperature. Extraction methods of the acetaldehyde-hydrazone (AcH-DNP) stable derivative and product stability studies were carried out. Acetaldehyde was identified by its retention time in comparison with AcH-DNP standard, using a new chromatography gradient program, and quantitated based on external reference standards and standard addition calibration curves in the presence and absence of ethanol. Derivatization of acetaldehyde was performed at pH 4.0 with an 80-fold molar excess of DNPH. The reaction was completed in 40 minutes at ambient temperature, and the product was stable for 2 days. A clear separation of AcH-DNP from DNPH was obtained with a new 11-minute chromatography program. Acetaldehyde detection was linear up to 80 μM. The recovery of acetaldehyde was >88% in culture media and >78% in plasma. We quantitatively determined the ethanol-derived acetaldehyde in hepatoma cells, rat blood and plasma with a detection limit around 3 μM. The accuracy of the method was <9% for intraday and <15% for interday measurements, in small volume (70 μl) plasma sampling. An optimized method for the quantitative determination of acetaldehyde in biological systems was developed using derivatization with DNPH, followed by a short RP-HPLC separation of AcH-DNP. The method has an extended linear range, is reproducible and applicable to small-volume sampling of culture media and biological fluids.

Pharmaceutical development and preliminary clinical testing of an oral solid dispersion formulation of docetaxel (ModraDoc001)

For use in chronic oral chemotherapeutic regimens, the potent anticancer drug docetaxel needs a solid oral dosage form. Because docetaxel has a very low permeability and a very low aqueous solubility (biopharmaceutical classification system class IV), a pharmacokinetic booster was combined with a newly developed solid dispersion formulation to improve the oral bioavailability of docetaxel. The best performing solid dispersion was a 1/9/1 w/w/w ternary mixture of docetaxel, polyvinylpyrrolidone (PVP)-K30 and sodium lauryl sulphate (SLS). In a phase I clinical trial, with ritonavir as pharmacokinetic booster, the docetaxel premix solution (TAXOTERE) was pharmacokinetically evaluated against the solid dispersion formulation filled into hard gelatin capsules (ModraDoc001 15 mg capsules). There were no significant differences between the pharmacokinetic parameters of docetaxel after administration of docetaxel premix solution or ModraDoc001 15 mg capsules, although there was a trend towards a higher and more variable exposure to docetaxel after oral administration of docetaxel premix solution (513 ± 219 vs. 790 ± 669 ng h/mL). The low inter-individual variability of docetaxel exposure (44%), the dosing accuracy, and the absence of ethanol and polysorbate are major advantages of ModraDoc001 15 mg capsules over docetaxel premix solution.

Mutant selection and phenotypic and genetic characterization of ethanol-tolerant strains of Clostridium thermocellum

Clostridium thermocellum is a model microorganism for converting cellulosic biomass into fuels and chemicals via consolidated bioprocessing. One of the challenges for industrial application of this organism is its low ethanol tolerance, typically 1-2% (w/v) in wild-type strains. In this study, we report the development and characterization of mutant C. thermocellum strains that can grow in the presence of high ethanol concentrations. Starting from a single colony, wild-type C. thermocellum ATCC 27405 was sub-cultured and adapted for growth in up to 50 g/L ethanol using either cellobiose or crystalline cellulose as the growth substrate. Both the adapted strains retained their ability to grow on either substrate and displayed a higher growth rate and biomass yield than the wild-type strain in the absence of ethanol. With added ethanol in the media, the mutant strains displayed an inverse correlation between ethanol concentration and growth rate or biomass yield. Genome sequencing revealed six common mutations in the two ethanol-tolerant strains including an alcohol dehydrogenase gene and genes involved in arginine/pyrimidine biosynthetic pathway. The potential role of these mutations in ethanol tolerance phenotype is discussed.

Groundwater Remediation Using an Enricher Reactor—Permeable Reactive Biobarrier for Periodically Absent Contaminants

A combined enricher reactor (ER)-permeable reactive biobarrier (PRBB) system was developed to treat groundwater with contaminants that appear in batches. An enricher reactor is an offline reactor used to enrich contaminant degraders by supplying necessary growth materials, and the enriched degraders are used to augment PRBB to increase its performance after a period of contaminant absence. Bench-scale experiments on PRBBs with and without bacterial supply from the enricher reactor were conducted to evaluate PRBB removal performances for benzene, which was used as a model contaminant. Benzene absence periods of 10 and 25 days were tested in the presence and absence of ethanol. The PRBBs without the bioaugmentation from the enricher reactor experienced a decrease in performance from approximately 65% to 30% after benzene reappeared. The presence of ethanol accelerated the benzene removal performance recovery of PRBBs. The 25-day benzene absence period caused greater changes in the bacterial community structure, regardless of the ethanol availability.

Arouser Reveals a Role for Synapse Number in the Regulation of Ethanol Sensitivity

A reduced sensitivity to the sedating effects of alcohol is a characteristic associated with alcohol use disorders (AUDs). A genetic screen for ethanol sedation mutants in Drosophila identified arouser (aru), which functions in developing neurons to reduce ethanol sensitivity. Genetic evidence suggests that aru regulates ethanol sensitivity through its activation by Egfr/Erk signaling and its inhibition by PI3K/Akt signaling. The aru mutant also has an increased number of synaptic terminals in the larva and adult fly. Both the increased ethanol sensitivity and synapse number of the aru mutant are restored upon adult social isolation, suggesting a causal relationship between synapse number and ethanol sensitivity. We thus show that a developmental abnormality affecting synapse number and ethanol sensitivity is not permanent and can be reversed by manipulating the environment of the adult fly.

Acute ethanol exposure prevents PMA-mediated augmentation of N-methyl- d-aspartate receptor function in primary cultured cerebellar granule cells

Many intracellular proteins and signaling cascades contribute to the ethanol sensitivity of native N-methyl- d-aspartate receptors (NMDARs). One putative protein is the serine/threonine kinase, protein kinase C (PKC). The purpose of this study was to assess if PKC modulates the ethanol sensitivity of native NMDARs expressed in primary cultured cerebellar granule cells (CGCs). With the whole-cell patch-clamp technique, we assessed if ethanol inhibition of NMDA-induced currents (I NMDA) (100 μM NMDA plus 10 μM glycine) were altered in CGCs in which the novel and classical PKC isoforms were activated by phorbol-12-myristate-13-acetate (PMA). Percent inhibition by 10, 50, or 100 mM ethanol of NMDA-induced steady-state current amplitudes (I SS) or peak current amplitudes (I Pk) of NMDARs expressed in CGCs in which PKC was activated by a 12.5 min, 100 nM PMA exposure at 37°C did not differ from currents obtained from receptors contained in control cells. However, PMA-mediated augmentation of I Pk in the absence of ethanol was abolished after brief applications of 10 or 1 mM ethanol coapplied with agonists, and this suppression of enhanced receptor function was observed for up to 8 min post-ethanol exposure. Because we had previously shown that PMA-mediated augmentation of I NMDA of NMDARs expressed in these cells is by activation of PKCα, we assessed the effect of ethanol (1, 10, 50, and 100 mM) on PKCα activity. Ethanol decreased PKCα activity by 18% for 1 mM ethanol and activity decreased with increasing ethanol concentrations with a 50% inhibition observed with 100 mM ethanol. The data suggest that ethanol disruption of PMA-mediated augmentation of I NMDA may be due to a decrease in PKCα activity by ethanol. However, given the incomplete blockade of PKCα activity and the low concentration of ethanol at which this phenomenon is observed, other ethanol-sensitive signaling cascades must also be involved.

Rapid discrimination and counterfeit detection of perfumes by an electronic olfactory system

This work describes a fast, simple and low cost procedure to discriminate counterfeit perfumes by means of a commercial electronic olfactory system (EOS) based on thin film metal oxide semiconductor (MOS). As proof, the authentic Coco Mademoiselle women perfume by Chanel perfumes, two counterfeit and five inspired perfumes were used. Two different methods for the perfume sample preparation, in absence and in presence of ethanol, were compared. The best results were obtained in absence of ethanol. In this case, the fingerprint of the authentic sample obtained with the EOS was very characteristic, showing distinctive sets of radar plots for each perfume sample. To extract the available information from the resistance ( R) change of each sensor, the classical feature selection ( R/ R 0) has been employed. The principal component analysis (PCA) placed the samples of the eight perfumes into well-defined groups. The counterfeit and inspired samples were also clearly detected by comparison of their respective fingerprints, with PCA placing these samples into well separated groups. Finally, the results have been contrasted by a triangle test performed by trained panelists familiarized with perfumes.

Ethanol Interferes with Gene Regulatory Networks of Embryonic Stem Cells

Gestational exposure to ethanol may promote abnormalities in the fetus, including growth retardation, facial dysmorphology and mental disability, under the umbrella of Fetal Alcohol Spectrum Disorders (FASD). We studied the interference of ethanol with normal embryonic development in embryonic stem (ES) cells. Mouse ES cells (E14Tg2A) were cultured in feeder‐free conditions, and converted to neural lineages in adherent monolayer in the presence or absence of ethanol. High throughput BioMark microfluidic 48.48 dynamic array chips (Fluidigm) measuring 2,304 real time quantitative PCR assays were used to assess levels of transcription factors and target genes involved in cell fate commitment during cell differentiation. Our data show that during ES cell differentiation i) genes regulating pluripotency and self‐renewal (Sox2, Nanog, Oct4, Klf4, Rex1, Dax1) were downregulated, iii) neuroectoderm and neural markers (e.g., Cxcl12, Nrip1) were upregulated, and ii) ethanol exposure delayed these changes. Network analysis points to nodes and edges of the gene regulatory networks perturbed by ethanol during early differentiation that may be responsible for aberrant neuroectodermal development in FASD.Supported by TJU Pilot Research Award, Dept Funds, and Graham Foundation.

Characteristics of SBA-15 Synthesized by One-Step Method

The synthesis of SBA-15 with or without ethanol using a one-step method and its porosity were investigated. Well-ordered SBA-15 with a rod like morphology could be obtained from starting mixtures by aging at a low (60 degrees C) or high (80 degrees C) temperature in the presence of an appropriate amount of ethanol. By contrast, mesoporous materials containing aggregates of particles obtained in the absence of ethanol at high aging temperatures (70 or 80 degrees C) had complex poorly ordered structures, indicating that ethanol can affect the structure of mesoporous materials. In addition, the poorly ordered mesoporous materials had unusual desorption characteristics, namely they possessed a two-step desorption branch.

Enzyme inactivation by ethanol and development of a kinetic model for thermophilic simultaneous saccharification and fermentation at 50 °C with Thermoanaerobacterium saccharolyticum ALK2

Studies were undertaken to understand phenomena operative during simultaneous saccharification and fermentation (SSF) of a model cellulosic substrate (Avicel) at 50°C with enzymatic hydrolysis mediated by a commercial cellulase preparation (Spezyme CP) and fermentation by a thermophilic bacterium engineered to produce ethanol at high yield, Thermoanaerobacterium saccharolyticum ALK2. Thermal inactivation at 50 °C, as shown by the loss of 50% of enzyme activity over 4 days in the absence of ethanol, was more severe than at 37 °C, where only 25% of enzyme activity was lost. In addition, at 50 °C ethanol more strongly influenced enzyme stability. Enzyme activity was moderately stabilized between ethanol concentrations of 0 and 40 g/L, but ethanol concentrations above 40 g/L accelerated enzyme inactivation, leading to 75% loss of enzymatic activity in 80 g/L ethanol after 4 days. At 37 °C, ethanol did not show a strong effect on the rate of enzyme inactivation. Inhibition of cellulase activity by ethanol, measured at both temperatures, was relatively similar, with the relative rate of hydrolysis inhibited 50% at ethanol concentrations of 56.4 and 58.7 g/L at 50 and 37 °C, respectively. A mathematical model was developed to test whether the measured phenomena were sufficient to quantitatively describe system behavior and was found to have good predictive capability at initial Avicel concentrations of 20 and 50 g/L.

Drosophila lacking a homologue of mammalian ALDH2 have multiple fitness defects

Little is known about the roles of aldehyde dehydrogenases in non-vertebrate animals. We recently showed that in Drosophila melanogaster, an enzyme with ∼70% amino acid identity to mammalian ALDH2 is necessary for detoxification of dietary ethanol. To investigate other functions of this enzyme, DmALDH, encoded by the gene Aldh, we compared two strains homozygous for Aldh-null mutations to two closely related wild type strains in measures of fitness and stress resistance in the absence of ethanol. Aldh-null strains have lower total reproductive rate, pre-adult viability, resistance to starvation, and possibly longevity than wild-type strains. When maintained under hyperoxia, Aldh nulls die more quickly and accumulate higher levels of protein carbonyls than wild-types, thereby providing evidence that DmALDH is important for detoxifying reactive aldehydes generated by lipid peroxidation. However no effect of Aldh was seen on protein carbonyl levels in flies maintained under normoxia. It is possible that Aldh nulls experience elevated rates of protein carbonylation under normoxia, but this is compensated (at a fitness cost) by increased rates of degradation of the defective proteins. Alternatively, the fitness defects of Aldh nulls under normoxia may result from the absence of one or more other functions of DmALDH, unrelated to protection against protein carbonylation.

Effect of Different Enological Practices on Skin and Seed Proanthocyanidins in Three Varietal Wines

Proanthocyanidins are important for wine quality since they participate in astringency, bitterness and color. Given the localization of proanthocyanidins in the berry (skin and seeds), different methods have been developed that help to modulate the release of these phenolic compounds. In this study, the effect of two low prefermentative temperature techniques (cold soak and must freezing with dry ice) and the use of macerating enzymes has been studied during the vinification of three different varietal wines (Monastrell, Syrah and Cabernet Sauvignon) to assess their influence on wine proanthocyanidin concentration and composition. Syrah wines showed the lowest proanthocyanidin content, together with the lowest mDP and the highest percentage of galloylation in its proanthocyanidins. Monastrell and Cabernet Sauvignon wines showed similar proanthocyanidin concentration. The application of the low temperature prefermentative maceration (cold soak) was the most effective treatment, increasing the proanthocyanidin concentration in Monastrell and Cabernet Sauvignon wines although neither of the treatments had any effect on Syrah wines. As regards the effect of the different treatments on the proanthocyanidin composition, the results seem to indicate that the observed increases were mainly due to an increase in seed proanthocyanidins, even in the case of cold soak treatments, which occur in the absence of ethanol, suggesting that ethanol is not so crucial in the extraction of seed proanthocyanidins.

Atomic-force microscopy of nickel nanoparticles possessing electrocatalytic properties

Surface morphology of highly oriented pyrolytic graphite with electrodeposited nickel nanoparticles was studied by atomic-force microscopy in the presence and absence of ethanol. Voltammetric and atomic-force microscopic data and histograms of particle size distribution were used to evaluate the unit-area catalytic activity of the nanocatalyst in relation to conditions of its preparation.

Ethanol-induced small heat shock protein genes in the differentiation of mouse embryonic neural stem cells

Neural stem cells (NSCs) of the neuroepithelium differentiate into one of three central nervous system (CNS) cell lineages: neurons, astrocytes, or oligodendrocytes. In this study, the differentiation potential of NSCs from the forebrain of embryonic day 15 (E15) mouse embryos was analyzed using immunocytochemistry. NSCs were differentiated early in the presence or absence of ethanol (50mM), and gene expression patterns among NSCs, differentiated cells and ethanol-treated differentiated cells were assessed by microarray and real-time PCR analysis. Genes that were up-regulated in differentiated cells both in the presence and in the absence of ethanol when compared to NSCs were related to the Wnt signaling pathway, including Ctnna1, Wnt5a, Wnt5b, Wnt7a, Fzd3, and Fzd2; genes related to cell adhesion, including Cadm1, Ncam1, and Ncam2; and genes encoding small heat shock proteins, including HspB2, HspB7, and HspB8. In particular, the expression levels of HspB2 and HspB7 were elevated in ethanol-treated differentiated cells compared to non-treated differentiated cells. The gene expression patterns of various heat shock transcription factors (HSFs), proteins that regulate the transcription of heat shock genes, were also analyzed. The expression levels of HSF2 and HSF5 increased in differentiated cells in the presence and absence of ethanol when compared to NSCs. Of these two genes, HSF5 demonstrated an enhanced up-regulation, particularly in ethanol-treated differentiated cells compared to cells that were differentiated in the absence of ethanol. These results imply that HspB2 and HspB7, which are small heat shock proteins with tissue-restricted expression profiles, might be up-regulated by ethanol during the short-term differentiation of NSCs.

Oxygen Reduction Reaction on Pt–NiO x /C, Pt–CoO x /C, and Pt–SnO2/C Electrodes in the Presence of Ethanol

Pt–NiO x /C, Pt–CoO x /C, and Pt–SnO2/C catalysts, prepared by thermal decomposition of polymeric precursors, are characterized for the oxygen reduction reaction in the presence of ethanol, and their tolerance to the presence of alcohol is determined. The electrocatalytic activities of Pt–NiO x /C, Pt–CoO x /C, and Pt–SnO2/C in the absence of ethanol are similar to those of the Pt/C catalysts or lower, depending on electrode composition. However, these catalysts present higher catalytic activity for the oxygen reduction reaction in the presence of ethanol, compared with the Pt/C catalyst. This makes Pt–NiO x /C, Pt–CoO x /C, and Pt–SnO2/C potentially applicable as cathodes in direct ethanol fuel cells, thereby reducing the effect of ethanol crossover.

Innate Immunity and Inflammation in Sepsis: Mechanisms of Suppressed Host Resistance in Mice Treated with Ethanol in a Binge-Drinking Model

Sepsis is a major cause of mortality worldwide. Acute or chonic ethanol exposure typically suppresses innate immunity and inflammation and increases the risk of mortality in patients with sepsis. The study described here was designed to address the mechanism(s) by which acute ethanol exposure alters the course of sepsis. Ethanol administered to mice shortly before Escherichia coli (injected ip to produce sepsis) decreased production of proinflammatory cytokines and chemokines for several hours. Bacteria in the peritoneal cavity decreased over time in control mice and were mostly cleared by 21 h, but in ethanol-treated mice, bacteria increased over time to more than 2 × 10(8) at 21 h. Killing of bacteria in macrophages and neutrophils was apparently compromised by ethanol, as the percentage of these cells that had cleared phagocytosed bacteria increased over time in control mice but not in ethanol-treated mice. The roles of TLR4, MyD88, and myeloperoxidase (MPO) were evaluated using mutant or knockout mice, and these experiments indicated that mice with hyporesponsive TLR4 survived better than those with normal TLR4. Lack of MyD88 or MPO did not significantly alter survival in the presence or absence of ethanol. Ethanol decreased survival in all groups. This indicates that the antimicrobial activities induced though TLR4 are dispensable for survival but contribute to lethality late in the course of sepsis. Thus, the effects of ethanol responsible for lethal outcome in sepsis are not dependent on inhibition of TLR4 signaling, as we and others had previously suspected.

GABA(A) receptors in the dorsal raphé nucleus of mice: escalation of aggression after alcohol consumption.

The dorsal raphé nucleus (DRN), the origin for serotonin (5-HT) in forebrain areas, has been implicated in the neural control of escalated aggression. Gamma aminobutyric acid type-A (GABA(A)) and type-B (GABA(B)) receptors are expressed in the DRN and modulate 5-HT neuronal activity, and both play a role in the behavioral effect of alcohol. The purpose of this study is to examine the interaction between drugs acting on GABA receptors in the DRN and alcohol in their effects on aggressive behaviors. Male CFW mice, housed with a female, were trained to self-administer ethanol (1.0 g/kg) or water via an operant conditioning panel in their home cage. Immediately after they drank either ethanol or water, the animals were microinfused with a GABAergic drug into the DRN, and their aggressive behaviors were assessed 10 min later. Muscimol (0.006 nmol), a GABA(A) receptor agonist, escalated alcohol-heightened aggression but had no effect in the absence of ethanol. This effect of muscimol was prominent in the animals that showed alcohol-heightened aggression, but not the animals that reduced or did not change aggressive behavior after ethanol infusion compared to water. On the other hand, the GABA(B) agonist baclofen (0.06 nmol) increased aggressive behavior similarly in both water and ethanol conditions. Antagonists of the GABA(A) and GABA(B) receptors, bicuculline (0.006 nmol) and phaclofen (0.3 nmol) respectively, did not suppress heightened-aggressive behavior induced by ethanol self-administration. GABA(A) receptors in the DRN are one of the neurobiological targets of alcohol-heightened aggression. Activation of the GABA(B) receptors in the DRN also produced escalated aggression, but that is independent of the effect of alcohol.

The effect of component of microemulsion for transdermal delivery of nicardipine hydrochloride

Purpose: Nicardipine hydrochloride has been used widely for the treatment of angina pectoris and hypertension. Because of its extensive first pass metabolism after oral administration, the transdermal administration of nicardipine microemulsions was developed in this study. Methods: Microemulsions consisted of isopropyl myristate (IPM), surfactant mixture of Tween 80/Span 80 and/or Tween 80/Span 20, co-surfactant (ethanol) and aqueous phase. Pseudo-ternary phase diagrams were constructed using water titration method. The effect of component of microemulsion on the percutaneous absorption of drug was evaluated by in vitro permeation study. Results: The area of microemulsion isotropic region in the presence of ethanol was comparably larger in the absence of ethanol. The mean droplet size of nicardipine microemulsions ranged from 70 to 123 nm. With addition of ethanol, the droplet size became smaller. The permeation rate and extent of nicardipine microemulsion transport across rat skin was affected by the components of microemulsion. Nicardipine microemulsion had higher flux at surfactant mixture with lower hyrophile-lipophile balance (HLB) value and Tween content. Conclusions: The microemulsion consisted of 52% IPM, 35% surfactant mixture and 13% water had higher permeation rate through rat skin above 122.53±1.87 μg/cm2/h and was expected to develop a transdermal delivery system.

Kinetics of the simultaneous oxidation of nickel(II) and sulfur(IV) by oxygen in alkaline medium in Ni(II)–sulfur(IV)–O2 system

AbstractIn the Ni(II)–S(IV)–O2 system in the region of pH &gt; 8.4, both Ni(II) and S(IV) are simultaneously autoxidized, and when sulfur is consumed fully NiOOH precipitates. At pH &gt; 8.4, ethanol has no effect on the rate, whereas ammonia strongly inhibits the reaction when pH &gt; 7.0. The kinetics of the reaction, in both the presence and the absence of ethanol, is defined by the rate law equation image where k is the rate constant, KO is the equilibrium constant for the adsorption of O2 on Ni(OH)2 particle surface. In ammonia buffer, the factor F is defined by equation image where K, KOH, K1, K2, K3, and K4 are the stability constants of NiSO3, NiOH+, Ni(NH3)2+, Ni(NH3), Ni(NH3), and Ni(NH3), respectively. In unbuffered medium, the factor F reduces to equation image The values of k and Ksp were found to be (1.3 ± 0.08) × 10−1 s−1 and (4.2 ± 3.5) × 10−16, respectively, at 30°C. A nonradical mechanism that assumes the adsorption of both SO32− and O2 on the Ni(OH)2 particle surface has been proposed. At pH ≤ 8.2, Ni(II) displays no catalytic activity for sulfur(IV)‐autoxidation and it is also not oxidized to NiOOH. © 2010 Wiley Periodicals, Inc. Int J Chem Kinet 42: 464–478, 2010

Molecular Mechanisms of Ethanol-Induced Pathogenesis Revealed by RNA-Sequencing

Acinetobacter baumannii is a common pathogen whose recent resistance to drugs has emerged as a major health problem. Ethanol has been found to increase the virulence of A. baumannii in Dictyostelium discoideum and Caenorhabditis elegans models of infection. To better understand the causes of this effect, we examined the transcriptional profile of A. baumannii grown in the presence or absence of ethanol using RNA-Seq. Using the Illumina/Solexa platform, a total of 43,453,960 reads (35 nt) were obtained, of which 3,596,474 mapped uniquely to the genome. Our analysis revealed that ethanol induces the expression of 49 genes that belong to different functional categories. A strong induction was observed for genes encoding metabolic enzymes, indicating that ethanol is efficiently assimilated. In addition, we detected the induction of genes encoding stress proteins, including upsA, hsp90, groEL and lon as well as permeases, efflux pumps and a secreted phospholipase C. In stationary phase, ethanol strongly induced several genes involved with iron assimilation and a high-affinity phosphate transport system, indicating that A. baumannii makes a better use of the iron and phosphate resources in the medium when ethanol is used as a carbon source. To evaluate the role of phospholipase C (Plc1) in virulence, we generated and analyzed a deletion mutant for plc1. This strain exhibits a modest, but reproducible, reduction in the cytotoxic effect caused by A. baumannii on epithelial cells, suggesting that phospholipase C is important for virulence. Overall, our results indicate the power of applying RNA-Seq to identify key modulators of bacterial pathogenesis. We suggest that the effect of ethanol on the virulence of A. baumannii is multifactorial and includes a general stress response and other specific components such as phospholipase C.