Genetic association between antipsychotic induced weight gain and FTO gene in a Sri Lankan population

Background: About 30% of patients treated with second generation antipsychotics (SGA) experience weight gain. Although there is evidence that the FTO gene is associated with obesity its role in antipsychotic induced weight gain is not so clear. Methods: A genetic association study was carried out to identify the association between FTO rs9939609 and antipsychotic induced weight gain. Sample consisted of 180 cases and 120 controls. Cases were patients diagnosed with schizophrenia or schizoaffective disorder, treated with second-generation antipsychotics for a minimum of 3 months, and had gained at least 10% of body weight. Controls were patients with schizophrenia treated with second-generation antipsychotics for a minimum of 3 months but had not gained ≥10% of body weight. Genomic DNA was extracted from whole blood. Polymerase chain reaction of the samples was done. Real-time quantitative PCR (qPCR) was carried out using BIO-RAD CFX96 Touch TM PCR detection system. Results: Females were significantly more among cases (58.3%) than controls (35%). Cases (52.4%) were significantly more likely to be overweight or obese than controls (13.8%). Genotype distribution was in Hardy-Weinberg equilibrium (p=0.43). Cochran-Armitage trend test was not significant. Risk of antipsychotic induced weight gain in the AA genotype [OR 1.69 (95% CI 0.74-3.86)] and AT genotype [OR 1.1 (95% CI 0.67-1.79)] were not significantly higher than the TT genotype. Recessive model showed that AA/AT genotypes were at significantly higher risk of being obese/overweight [OR 1.84 (95% CI 1.05-3.2)]. Conclusions: There was no significant association between FTO rs9939609 and antipsychotic induced weight gain. AA/AT genotypes had significantly higher risk of overweight/obesity.

Cytokine expression in response to root repair agents.

AIM: To evaluate the expression of TNF-α, IL-6, IFN-γ, TGF-ß, IL-4, IL-10, RANKL, RANK and OPG on mouse calvarial bone treated with MTA, Geristore® and Emdogain® . METHODOLOGY: Bone wounds were made on the heads of C57BL/6 mice, breaking the periosteum and the cortical surface of the calvaria. Each repair agent was inserted into sectioned Eppendorf microtubes and placed on the bone wound, and soft tissues were sutured. At 14 and 21 days, animals were sacrificed and the treated region was dissected. The calvaria bone was removed, and RNA was extracted. mRNA expression of the aforementioned cytokines was assessed using real-time PCR. Data were analysed by nonparametric methods, including the Mann-Whitney and Kruskal-Wallis tests (P < 0.05). RESULTS: Following treatment with Emdogain® and MTA, mRNA expression of RANKL, RANK and OPG increased significantly (P < 0.05) between days 14 to 21. Geristore® did not alter the basal expression of these mediators during the same period of evaluation. Whilst treatment with Emdogain® did cause a significant increase in TNF-α mRNA expression between days 14 and 21 (P < 0.05), treatment with MTA did not alter the basal expression of this cytokine at either experimental time point. However, TNF-α mRNA expression was down-regulated significantly at day 21 (P < 0.05) when Geristore® was applied. A significant increase in the mRNA expression of IL-6, TGF-ß, IL-10, IL-4 and IFN-γ was observed with Emdogain® and MTA treatment between days 14 to 21, whereas Geristore® reduced significantly the expression of IL-6, TGF-ß and IL-4 (P < 0.05). CONCLUSION: The clinical indication of these repair agents depends on the root resorption diagnosis. Whilst MTA and Emdogain® induce a pro- and anti-inflammatory response early and late, respectively, Geristore® was not associated with an inflammatory reaction when compared with both repair agents.

Synthetic Organotellurium Compounds Sensitize Drug-Resistant Candida albicans Clinical Isolates to Fluconazole.

Invasive Candida albicans infections are a serious health threat for immunocompromised individuals. Fluconazole is most commonly used to treat these infections, but resistance due to the overexpression of multidrug efflux pumps is of grave concern. This study evaluated the ability of five synthetic organotellurium compounds to reverse the fluconazole resistance of C. albicans clinical isolates. Compounds 1 to 4, at <10 µg/ml, ameliorated the fluconazole resistance of Saccharomyces cerevisiae strains overexpressing the major C. albicans multidrug efflux pumps Cdr1p and Mdr1p, whereas compound 5 only sensitized Mdr1p-overexpressing strains to fluconazole. Compounds 1 to 4 also inhibited efflux of the fluorescent substrate rhodamine 6G and the ATPase activity of Cdr1p, whereas all five of compounds 1 to 5 inhibited Nile red efflux by Mdr1p. Interestingly, all five compounds demonstrated synergy with fluconazole against efflux pump-overexpressing fluconazole-resistant C. albicans clinical isolates, isolate 95-142 overexpressing CDR1 and CDR2, isolate 96-25 overexpressing MDR1 and ERG11, and isolate 12-99 overexpressing CDR1, CDR2, MDR1, and ERG11 Overall, organotellurium compounds 1 and 2 were the most promising fluconazole chemosensitizers of fluconazole-resistant C. albicans isolates. Our data suggest that these novel organotellurium compounds inhibit pump efflux by two very important and distinct families of fungal multidrug efflux pumps: the ATP-binding cassette transporter Cdr1p and the major facilitator superfamily transporter Mdr1p.

HydroCalc Proteome: a tool to identify distinct characteristics of effector proteins.

Bacterial pathogenicity is associated with secretion of effector proteins into intra- and extracellular spaces. These proteins interfere with cellular processes such as inhibition of phagosome-lysosome fusion, induction of apoptosis and autophagy, activation and suppression of kinases, regulation of receptor activity, and modulation of transcription factors. Knowledge regarding the characteristics of these proteins would assist in pathogenicity studies, and help to identify possible and novel targets for antibacterial drugs. Amino acid hydropathy is a property that can affect behavior patterns in effector proteins. The HydroCalc Proteome tool analyzes total hydropathy, average hydropathy, C-terminal hydropathy, C-terminal load, and basic polar amino acids at the C-terminus. These five properties could contribute to the identification of proteins with an effector potential. HydroCalc Proteome is a web tool that provides a simple interface for the analysis of hydropathy properties in proteins. This tool permits the analysis of a single protein or even the complete proteome, which cannot be achieved by using other hydropathy tools. The tool displays the result of five properties related to effector proteins in a single table. The HydroCalc Proteome (www.gmb.bio.br/hydrocalc) is a powerful tool for protein analysis, and can contribute to the study of effector proteins.

Validity of the Sinhala version of the centre for epidemiological studies depression scale (CES-D) in out-patients.

OBJECTIVES: To translate and validate the Sinhala version of the Centre for Epidemiological Studies Depression scale (CES-D) for diagnosing depression in out-patients. DESIGN: A combined qualitative and quantitative approach was used for the translation of the CES-D. Sample size was calculated to detect a targeted sensitivity and specificity of 85%. The sample consisted of 75 participants diagnosed with major depressive disorder according to DSM IV criteria and 75 gender matched controls. Criterion validity was assessed using receiver operating charact-eristic (ROC) analysis. The Structured Clinical Interview for DSM-IV (SCID-II) conducted by a psychiatrist was used as the gold standard. RESULTS: Mean age of the sample was 33 years. There were 91 females (60.7%). There was significant difference in the mean CES-D scores between cases (13.94) and controls (6.58) [t=14.50, df=148, p<0.001]. A score of ≥ 16 gave a sensitivity of 84% and specificity of 92%. A score of ≥ 21 gave a sensitivity of 73.3% and specificity of 96%. The Cronbach's alpha was 0.93. The four items that were reverse coded had poor correlation with total scores. The average correlation coefficient for the reverse-scored items was 0.35 and for the rest of the items 0.63. Principal component analysis with oblique rotation identified four factors. Factor 1 corresponds to the "depressed affect" and "somatic complaints" in the original model proposed by Radloff. Factor 2 corresponds to the interpersonal concerns. Factors 3 and 4 loaded the reversed coded items. CONCLUSIONS: The Sinhala version of the CES-D is a valid and reliable instrument for diagnosing major depressive disorder.

Lack of genotoxicity in Astyanax bimaculatus and Oreochromis niloticus of 17α-methyltestosterone used in fish hatcheries to produce male monosex populations.

17α-Methyltestosterone (MT) is widely used in fish hatcheries of many countries to produce male monosex populations. Its genotoxic risk to fish species is not well known and studies in other in vivo models are still inconclusive. MT was tested for genotoxicity in the fish species Oreochromis niloticus (tilapia), a target species, and Astyanax bimaculatus (lambari), a native non-target species. Genotoxicity was evaluated by the micronucleus test (MN), nuclear abnormalities (NA), and comet assay using peripheral erythrocytes of both species after a 96-h exposure to MT at concentrations of 0.01, 0.1, and 1.0 mg/L in the water. At the lowest exposure level of 0.01 mg/L, MT induced MN in both species and NA only in O. niloticus. These effects were not observed in the comet assay. Chromatographic analysis of water samples collected from aquariums at the beginning and end of each experiment showed that MT was consumed during the 96-h exposure. At the highest level of exposure (1.0 mg/L), 81.69% of the hormone was consumed during the exposure period. The chromatogram showed that at the lowest concentration level of 0.01 mg/L, 99.56% MT was consumed by the end of the exposure period. Thus, exposure to MT did not cause genotoxicity in either fish species.

Geographical variation in admissions due to poisoning in Sri Lanka: a time series analysis.

OBJECTIVES: To carry out time series analyses of hospital admissions for poisoning between 1995-2008 in all districts in Sri Lanka to identify trends and geographical variations in the substances used in poisoning. METHODS: Data of hospital admissions from 1995-2008 due to poisoning were obtained from the Annual Health Bulletins published by the Ministry of Health. Data were converted to annual rates per 100,000 population. Time trends in the rates of suicide and self-poisoning were calculated using univariate time series analysis. RESULTS: All districts except Kilinochchi and Mullaitivu showed an increase in the rates of admissions due to poisoning with drugs, medicaments and biological substances. Colombo, Hambantota, Kalutara and Anuradhapura showed an exponential increase. Hambantota, Monaragala, Nuwara Eliya and Colombo show an increase in the rate of admissions after pesticide poisoning. All other districts showed a linear decrease. Admissions due to all types of poisoning showed a negative trend in Anuradhapura, Polonnaruwa, Ampara, Matale and Batticoloa districts. Other districts show a positive trend in the rate of admissions for all types of poisoning. CONCLUSIONS: Results should be viewed with caution because they are based on analysis of secondary data. Although the rate of suicides has reduced since 1995, admissions due to self poisoning have increased in almost all districts. While pesticide poisoning is becoming less, there is a gradual shift to the use of drugs and medicaments in self poisoning. Poisoning with drugs, medicaments and biological substances are increasing both in urban and rural areas.

Early malnutrition favours heavy ethanol intake in weaning rats without long-lasting effects.

INTRODUCTION: Although food restriction is well known to increase ethanol intake, the subject has not been extensively studied in developing animals which could be more vulnerable to long-lasting effects. Therefore, the aim of this study was to show some findings concerning this subject. MATERIALS AND METHODS: Food restriction was used to produce malnutrition either during lactation (lactating dams food restricted 60%) or after weaning (pups food restricted 60%). At weaning, day 25, males were assigned to one of the following groups: C, food ad libitum throughout the experiment - control group; MW, malnourished only after weaning; ML, malnourished only during lactation period; and MLW, malnourished throughout the experiment, during lactation and after weaning. All rats were kept isolated in cages in which they could choose to drink either a 10% ethanol solution or tap water (from days 25 to 45). Re-exposure to this model was performed on day 49. Between exposure and re-exposure, rats drank tap water for 4 days. RESULTS: There was a significant effect of malnutrition during lactation, up to day 35, with heavy drinking patterns (ethanol intake day 2: C, 8 g/kg; MW, 9 g/kg; ML, 19 g/kg; and MLW, 22 g/kg). This heavy drinking pattern lasted until the recovery of body weight. Food restriction after weaning had significant effects after 14 days, when a statistically significant decrease in body weight occurred (body weight day 39: C, 147.8 g; MW, 98.5 g). Only rats which were persistently malnourished (MLW and MW) drank ethanol more significantly than their ad libitum-fed counterparts during the re-exposure period (ethanol intake: malnourished, 5 g/kg; and well-nourished, 2.5 g/kg). Adulteration of the ethanol solution with quinine (0.1 g/l) precluded the effect of malnutrition. CONCLUSIONS: Malnutrition during early development had no long-lasting effects on ethanol consumption. In addition, malnutrition increased ethanol consumption as long as it kept body weight low, which was apparently more significant in young animals.

Phytotherapy in reducing glycemic index and testicular oxidative stress resulting from induced diabetes: a review.

Oxidative stress plays a main role in the development of diabetes complications. The impairment of gonadal antioxidant potential and endocrine disturbance in diabetic males causes testicular damage and failure in sperm production. Plants have been widely used to control diabetes due their hypoglycemic and antioxidant potential, contributing towards the recovery of testicular function. Current study comprises a review of the literature on the main medicinal plants used in the recovery of testicular oxidative damage in animals with experimental diabetes. Eighteen plant species in the nineteen studies selected from the search strategy were evaluated. Plant extracts were evaluated according to their effects on blood glucose and insulin levels, antioxidant enzymes and oxidant levels, lipid peroxidation, total protein, testosterone levels, gonadosomatic index, diameter of seminiferous tubules, seminiferous epithelium height and integrity, number of germ cells at stage VII and apoptosis in the seminiferous epithelium, sperm production, motility, viability and morphology. After the analysis of the studies, it was observed that plant species, used alone or in combination, may control testicular oxidative damage triggered by diabetes. The antioxidant potential varies among species, with some plants proving to have a better performance in the recovery of reproduction parameters than others.

Correção de fratura Salter-Harris tipo I em porquinho-da-índia (Cavia porcellus) - relato de caso / [Correction of Salter-Harris type 1 fracture in guinea pig (Cavia porcellus) - case report]

Um filhote de porquinho-da-índia (Cavia porcellus) foi recebido para atendimento após histórico de ataque por cão. Na avaliação física, observou-se edema, dor e crepitação em membro pélvico direito, sugestivo de fratura. Na avaliação radiográfica, confirmou-se fratura Salter-Harris tipo I em epífise distal da tíbia. A resolução cirúrgica escolhida foi a associação de pino transarticular e coaptação externa com tala de Altman. O paciente teve acompanhamento radiográfico semanal e obteve alta médica no 35o dia de pós-cirúrgico, quando se observou consolidação com completo remodelamento ósseo.(AU)

A guinea pig (Cavia porcellus) cub presented edema, pain, and crepitus in the right pelvic limb after being attacked by a dog. Radiographic examination revealed Salter-Harris type 1 fracture on the distal region of the tibia. The surgery technique to correct the fracture involved an association of transarticular pinning and external coaptation with Altman splint. After surgery, radiographs of the patient were performed weekly and on the 35th post-surgery day, the bone was completely remodeled and healed, and the animal was dismissed.(AU)

Effect of two different types of malnutrition on the rate of elimination of ethanol in rats.

Rats were fed "3% casein" or a "calorie deficient" diet, in the form of commercial pellet diet (SDS) at 50% of the amount consumed by the control group, which was fed SDS pellets ad libitum. Both of the deficient groups showed failure of weight gain in comparison with the control group. Blood levels of ethanol were measured for 3 hr after intraperitoneal injection of 1 or 1.5 g/kg at 15, 29 and 36 days after commencement of the diet. In addition the calorie deficient group was studied immediately after feeding as well as in the fasting state. Blood levels of ethanol were measured and the apparent volume of distribution and rate of removal of ethanol from the blood were calculated. A rate of ethanol metabolism/g of liver was derived. The rate of removal of ethanol was markedly decreased in the 3% casein group to less than half of control values. Three hours after injection of ethanol circulating levels were less than 50 mg/100 ml in the control and calorie deficient groups but over 200 mg/100 ml in the group fed protein deficient diets. There were no major changes in volume of distribution and the only explanation for the finding is that there is a failure of ethanol metabolism in the rats fed the low protein diet. The implication is that protein deficient human populations who often consume considerable quantities of ethanol may have a high level of tissue exposure to ethanol though the rate of metabolite formation may be low.

Developmental toxicity of in utero exposure to toluene on malnourished and well nourished rats.

The aim of the present study was to investigate the effects of toluene on fetal development in well nourished and malnourished rats. Long-term behavioural consequences after in utero exposure were also studied. Toluene (1.2 g/kg s.c.) was administered daily to well nourished and to malnourished (food restricted to 50% of ad libitum intake) pregnant rats, during the second (8-15 days) or the third week of pregnancy (14-20 days). Offspring were evaluated for malformations, development of the skeleton, prenatal growth of the brain and liver, postnatal growth and long lasting behavioural effects. In utero exposure to toluene during the third week of pregnancy resulted in low body weight at birth, which persisted in the male offspring into adulthood. Malnutrition increased fetal susceptibility to the effects of toluene as indicated by evaluation of the development of the skeleton. Behavioral tests performed when the pups were 30 and 90 days old showed effects of in utero malnutrition (increased ambulation and worse performance in a shuttle box), but no behavioural effects related to toluene exposure were detected. These data indicate that in utero exposure to toluene can have long lasting effects on body growth and that maternal malnutrition increases the risk for toluene fetotoxicity.

Neurobehavioural development of the golden hamster.

The aim of this study was to establish a developmental profile for the golden hamster by using a systematic sequence of test procedures. One experimentally naive litter was tested each day from 0 to 25 days of postnatal age. The appearance of developmental landmarks (physical features and reflexes), spontaneous behaviour in an open field, homing behaviour and rota rod performance were studied. Infant mortality through infanticide was recorded in undisturbed and tested hamsters. The results indicated that most of the tests employed in the present study can be applied usefully in the evaluation of the neurobehavioural development of the golden hamster. The developmental profile for this species is described in detail. In comparison to rats and mice, hamsters display accelerated development of a number of characteristics, most notably incisor eruption and vaginal opening. Infanticide, the most troublesome problem in studies in which hamster litters must be disturbed, did not occur after day 3. As most reflexes and sensory abilities develop after this age, hamster pups can be used successfully in behavioural teratology evaluation.

A Portuguese version of the Christo Inventory for Substance-Misuse Services: a simple outcome evaluation tool.

The Christo Inventory for Substance-Misuse Services (CISS) is a single page outcome evaluation tool completed by drug alcohol service workers either on the basis of direct client interviews or of personal experience of their client supplemented by existing assessment notes. It was developed to assist substance misuse services to empirically demonstrate the effectiveness of their treatments to their respective funding bodies. Its 0 to 20 unidimensional scale consists of 10 items reflecting clients' problems with social functioning, general health, sexual/injecting risk behavior, psychological functioning, occupation, criminal involvement, drug/alcohol use, ongoing support, compliance, and working relationships. Good reliability and validity has already been demonstrated for the CISS [Christo et al., Drug and Alcohol Dependence 2000; 59: 189-197] but the original was written in English and a Portuguese version is presented here. The present review explores its applicability to a Brazilian setting, summarizes its characteristics and uses, and describes the process of translation to Portuguese. A pilot study conducted in a substance misuse service for adolescents indicated it is likely to be suitable for use among a Brazilian population. The simplicity, flexibility and brevity of the CISS make it a useful tool allowing comparison of clients within and between many different service settings.

Preliminary data on the prevalence of psychiatric disorders in Brazilian male and female juvenile delinquents.

The aim of the present investigation was to study the prevalence of psychiatric disorders in a sample of delinquent adolescents of both genders and to compare the prevalence between genders. A total of 116 adolescents (99 males and 17 females) aged 12 to 19 on parole in the State of Rio de Janeiro were interviewed using the screening interview based on the Schedule for Affective Disorders and Schizophrenia for School-Age Children -- Present and Lifetime (KSADS-PL). Data were collected between May 2002 and January 2003. Of 373 male and 58 female adolescents present in May 2002 in the largest institution that gives assistance to adolescents on parole in the city of Rio de Janeiro, 119 subjects were assessed (three of them refused to participate). Their average age was 16.5 years with no difference between genders. The screening interview was positive for psychopathology for most of the sample, with the frequencies of the suggested more prevalent psychiatric disorders being 54% for attention-deficit/hyperactivity disorder, 77% for conduct disorder, 41% for oppositional defiant disorder, 57% for anxiety disorder 57, 60% for depressive disorder 60, 63% for illicit drug abuse, and 58% for regular alcohol use. Internalizing disorders (depressive disorders, anxiety disorders and phobias) were more prevalent in the female subsample. There was no significant difference in the prevalence of illicit drug abuse between genders. There were more male than female adolescents on parole and failure to comply with the sentence was significantly more frequent in females. The high prevalence of psychopathology suggested by this study indicates the need for psychiatric treatment as part of the prevention of juvenile delinquency or as part of the sentence. However, treatment had never been available for 93% of the sample in this study.

Protein-energy malnutrition increases teratogenicity of hypervitaminosis A in rats.

The present study was designed to investigate the embryo-fetotoxicity of vitamin A in protein-energy malnourished animals. Retinyl palmitate (66, 99 and 132 mg/kg) suspended in corn oil was given by gavage to well-nourished and malnourished rats from gestational days 8 to 10 and cesarean sections were performed on day 20. All fetuses were weighed and examined for malformations before being prepared for skeletal evaluation. The proportion of malformed fetuses was higher in the malnourished group at each one of the three dose levels. The data indicate that malnourished animals are more susceptible to the toxic effects of retinyl esters.

Effects of toluene exposure during gestation on neurobehavioral development of rats and hamsters.

Pregnant rats and hamsters were exposed to toluene vapor (800 mg/m3) 6 h daily from gestation days 14 to 20, and 6 to 11, respectively. Growth, neuromotor development and performance of the offspring in behavioral tasks were assessed. In rats, toluene exposure increased the number of litters with low birth weight pups. Male rat offspring exposed to toluene displayed shorter latencies than male controls to choose one side of a T maze in a spontaneous alternation test. Hamsters exposed to toluene performed worse in a rotating rod test. These results confirm toluene fetotoxicity in rats and suggest an effect on exploratory behavior which may be related to hormonal changes in early life. Neuromotor effects of exposure of hamsters to toluene in utero deserve further investigation.

Ethanol elimination by rats as a function of reproductive state, gender and nutritional status.

Alcohol elimination was studied in rats of different ages, reproductive states and nutritional deprivation, with the following results: 1) blood levels of ethanol 180 min after a single dose of 1.5 g/kg, ip were significantly higher in adult male (74 days old, N = 5) than in young male rats (34 days old, N = 5): 92.4 +/- 8.4 vs 6.8 +/- 3.4 mg/100 ml, means +/- SD, respectively; 2) when male rats were given a low protein diet for 48 h, blood ethanol levels after a single dose were significantly increased in young males (38.6 +/- 14.6 mg/100 ml) but no effect after a single dose was found in the same animals at an older age (93.2 +/- 5.0 mg/100 ml); 3) blood levels in female rats were higher than in young males both in the virgin and pregnant states, but during lactation a significant drop in blood levels of ethanol was observed. Blood levels of ethanol (mg/100 ml) 180 min after a single dose of 1.5 g/kg, ip, in females, were: virgin (N = 6): 44.9 +/- 16.1, pregnant (N = 5): 40.0 +/- 10.4, lactant (N = 5) 8.8 +/- 5.8. This difference between virgin and pregnant and lactant rats was not related to changes in ADH activity which did not differ between groups. The present study indicates that in male rats the effect of a short-term protein deprivation on ethanol elimination is dependent on the age of the animal. In females, reproductive state is an important factor in determining ethanol elimination.

Can calories from ethanol contribute to body weight preservation by malnourished rats?

Our objective was to compare the use of calories from ethanol by well-nourished and malnourished rats in terms of body weight. Female Wistar rats weighing 170-180 g at the beginning of the study were used. The animals were divided into two groups (N = 12 each): group W received water ad libitum and group E an ethanol solution ad libitum as the only source of liquid throughout the experiment. The concentration of ethanol was increased weekly from 0 to 5, 10, 20 and 40% (v/v). In the well-nourished phase (A), all rats received food ad libitum (AW and AE). Ethanol treatment (AE) was then interrupted and water was offered to both groups. After 2 weeks both AW and AE rats were submitted to food restriction (50% of group AW food consumption), thus initiating the malnutrition phase (M). Liquid was offered as described before to the same W (MW) and E (ME) groups. The weight gain during the 1-week treatment of AE rats was similar to that of AW animals only when AE rats received the 5% (v/v) ethanol solution (9.16 vs 10.47 g). Weight loss was observed after exposure to 10% ethanol (P < 0.05) in spite of maintenance of caloric intake. Malnourished rats presented weight loss, which was attenuated by ethanol intake up to the 20% (v/v) solution and was related to an increased caloric offer. This effect was not observed with the 40% ethanol solution (-9.98 g). These data suggest that calories from ethanol were used to maintain body weight up to the concentration of 10% (v/v) (well-nourished) and 20% (v/v) (malnourished) and that ethanol has a toxic profile which depends on nutritional status.