Ethanol-derived Calories Diet Research Articles

Prevention by a Silymarin/Phospholipid Compound of Ethanol-Induced Social Learning Deficits in Rats

We explored the possibility that silymarin (SY), a fraction from Silybum marianum, might protect against the effects of in utero exposure to ethanol upon subsequent social memory function. Three groups of 8 pregnant female Sprague-Dawley rats each were provided with a liquid diet containing 35% ethanol derived calories (EDC). One experimental group received a daily subcutaneous injection of 400 mg/kg SY, the second, a 400 mg/kg oral dose of SY, a third group was maintained on the 35% EDC diet. A fourth group served as the pair-fed control group. The liquid diet regimen was maintained throughout pregnancy. Rats pups were fostered by dams in a fifth group that had been maintained on rat chow. At 90 days the pups were tested for social memory. Social recognition scores recorded for the ethanol pups were significantly poorer than those observed in both SY/ethanol groups and the chow group.

A Morphometric Analysis of Early Effects of Ethanol on Hepatocyte Organelles from Peromyscusmaniculatus

Abstract Pathological changes of hepatocytes from rats fed a 30% ethanol-derived calories diet for three weeks include noticeable ultrastructural changes including steatosis and hypertrophy of the smooth endoplasmic reticulum. We sought to examine hepatocytes of deer mice administered ethanol in an inhalation chamber for two weeks to determine if subtle changes occur in hepatocyte organelles prior to steatosis. Two strains of Peromyscus maniculatus, ADH-positive possessing hepatic cytosolic alcohol dehydrogenase and ADH-negative deer mice lacking this enzyme were purchased from the Peromyscus Genetic Stock Center (Univ. of South Carolina). They tested negatively for Hanta viruses. A morphometric analysis of the ultrastructure of ADH+(n=14) and ADH- (n=14) controls as well as experimentals exposed to chronic, intoxicating levels of ethanol was conducted. Blood ethanol levels were maintained between 1.25-1.75 mg/ml for two weeks in the experimentals.

Comparison of manual and morphometric analysis techniques for rat hepatocyte peroxisomes

It has been well documented that ethanol is toxic to hepatocytes and that peroxisomes are key organelles in the metabolism of ethanol. Lieber (1985) has confirmed that when rats are fed a 30% ethanol-derived calories diet for three weeks there are noticeable ultrastructural changes such as the accumulation of lipid (steatosis) and the hypertrophy of the smooth endoplasmic reticulum. We sought an efficient costeffective technique for obtaining reliable quantitative documentation of subtle changes within organelles of hepatocytes after rats were fed a 30% ethanol-derived calories diet for two weeks. One hundred micrographs of match paired ethanol-fed rats (n = 5) were compared with 100 micrographs of controls (n = 5) and analyzed by the point counting method; with a digital planimeter using SigmaScan/Image software (Jandel Inc.) and calculations were made using Excel v. 4.0 (Microsoft Corp.); as well as with Optimas image analysis software (Bioscan). Micrographs were taken with a Zeiss EM 10 transmission electron microscope.

Paternal alcohol exposure: paradoxical effect in mice and rats

Male mice consumed liquid alcohol diets containing 25%, 10% or 0% ethanol-derived calories (EDC). Animals receiving the 10 and 0% EDC diets were pair fed to those consuming the 25% EDC diet. After 7 or 14 weeks of consumption, males were bred to non-treated females. Offspring were tested for swimming behavior at 75 days of age. Offspring sired by alcohol-consuming males were more immobile regardless of duration of paternal alcohol consumption or housing conditions (group or isolated). Imipramine (5, 15 mg/kg) reversed this effect such that after drug treatment, alcohol-sired offspring were less immobile than controls. Propranolol (1, 3 mg/kg) eliminated the group differences. Yohimbine (1, 10 mg/kg) significantly increased immobility in all groups but did not reverse the effects of paternal alcohol consumption. Metergoline (1 mg/kg) increased immobility in all groups but did not reverse the effects of paternal alcohol exposure. When rat offspring of alcohol-consuming fathers were tested, their swimming behavior was opposite to that of mice (i.e., rats were less immobile). However, as observed in mice, the effect was reversed by imipramine. These results extend the evidence for paternally mediated behavioral mutagenesis. They also indicate that the direction of induced changes are species dependent and that some of these effects, regardless of direction of change, are mediated by neurochemical changes sensitive to imipramine.

Paternal alcohol consumption: effects on ocular response and serum antibody response to Pseudomonas aeruginosa infection in offspring.

Male Swiss-Webster mice that consumed liquid alcohol diets containing 25, 20, 15, 10, 5, or 0% ethanol-derived calories (EDC) for 7 weeks were bred to untreated females that were not exposed to alcohol diets. Males receiving the 20-0% EDC diets were pair-fed to those consuming the 25% EDC diet. At approximately 60 days of age, male offspring were challenged with 10(8) colony forming units of Pseudomonas aeruginosa onto the scarified cornea of one eye. The ocular response was then evaluated macroscopically for 4 weeks. Male offspring sired by alcohol-consuming fathers exhibited more severe ocular infection at 24 hr postinfection compared with mice sired by control fathers and, after 8 days postinfection, more corneas of the 25% EDC-sired progeny perforated than did the other groups. At 30 days after infection, serum immunoglobulin titers (IgM, IgG, IgA) specific to P. aeruginosa were determined by ELISA. Although the majority of the mice were unable to restore corneal clarity within 30 days postinfection, a strong serum IgG response was detected in pooled sera from those animals tested. Offspring were reinfected in the contralateral control eye at 30 days postprimary infection. Most control animals were able to restore corneal clarity in the contralateral control eye within 30 days postinfection, but less than a third of the alcohol-sired offspring did so. Again, the pooled sera that was tested indicated a strong humoral response, despite differences in corneal clarity. These studies indicate an increased susceptibility to infection and an impairment in restoration of corneal clarity in offspring sired by alcohol-consuming males, which does not appear to be mediated by serum antibody mechanisms.

Paternal alcohol consumption: effects of age of testing and duration of paternal drinking in mice.

Male mice consumed liquid alcohol diets containing 25, 20, 15, 10, 5, or 0% ethanol-derived calories (EDC). Animals receiving the 20-0% EDC diets were pair-fed to those consuming the 25% EDC diet. After 7 weeks of consumption males were bred to nontreated females. Offspring were tested for activity at 16-20 and 75 days of age. Offspring sired by alcohol-consuming males did not differ from controls in litter size, birth weight, or weight at weaning, but were less active than controls on several measures of activity. Many of these decreases were best defined in terms of linear trends. However, these differences were evident only for animals tested prior to 20 days of age for most activity measures. In a second experiment adult males continued to consume alcohol for another 7 weeks and were bred again. Offspring of this second breeding were tested for activity at 16 days of age and were compared with offspring sired by the same father from the previous breeding. Offspring sired after this longer duration of paternal alcohol consumption did not differ significantly from controls in any of the above-mentioned variables.

T lymphocyte populations in fetal alcohol syndrome.

There are reports that fetal alcohol syndrome (FAS) is associated with immune deficiency or DiGeorge syndrome. To investigate the effect of prenatal alcohol exposure on the immune system, we used a mouse model of FAS in which C57BL/6J female mice were fed a complete liquid diet containing 25% ethanol-derived calories (EDC) from gestational day (g.d.) 1 to 18. Thymus cell numbers were markedly reduced in 18-day fetuses exposed to ethanol. Thymocytes from fetuses from the 25% EDC diet group and from pair-fed and ad-libitum control diet groups were compared by flow cytometry for expression of T cell differentiation antigens. The proportions of L3T4- and Lyt-2 positive thymus cells were significantly reduced in alcohol-exposed fetuses compared to controls; however, the number of Thy-1-positive cells did not differ among any of the groups. Six-day old neonates exposed prenatally to ethanol from g.d. 1 to 13 had thymus and spleen T cell populations similar to those of controls in almost all cases, indicating a "catch-up" of T cell numbers in most animals. Spleen T cell function, assessed by response to Concanavalin A (Con A), or Con A plus T cell growth factors, was somewhat depressed in ethanol-exposed 6-day pups.

Effects of prenatal ethanol exposure in C57BL mice on locomotor activity and passive avoidance behavior

The purpose of this study was to examine the long-term behavioral effects of prenatal ethanol exposure in C57BL mice. Pregnant mice received free access to a liquid diet containing 25% ethanol-derived calories (EDC) from gestation days 6 to 18. Control animals were pair-fed an isocaloric 0% EDC diet during the same period of time. An additional control group was included that was maintained on standard lab chow and water throughout pregnancy. At 30 days of age, female offspring were tested for spontaneous locomotor activity in an open field under two lighting conditions (dim or bright illumination). Male offspring were tested in a passive avoidance task at 25 days of age. The activity results demonstrated that the 25% EDC female progeny were more active than controls. This hyperactivity was observed under both lighting conditions, despite the fact that all groups evidenced suppressed activity when tested under bright lights. With regard to passive avoidance behavior, male EtOH-exposed offspring required a greater number of trials to reach criterion than controls. Additionally, they exhibited shorter latencies to enter the shock-associated chamber after receiving a single shock. Taken together, these results confirm our previous findings and demonstrate that C57BL mice are sensitive to both the deleterious behavioral and morphological consequences of prenatal ethanol exposure.

Absence of differential motoric and thermic responses to clonidine in young rats exposed prenatally to alcohol

We studied the involvement of altered noradrenergic function in the behavioral consequences of fetal ethanol exposure. Pregnant rats were fed a liquid diet containing 35% Ethanol-Derived Calories (EDC), pair-fed a 0% EDC diet with sucrose substituted isocalorically for ethanol, or fed ad lib lab chow. In Experiment 1, offspring from these prenatal treatment groups were injected with the alpha-adrenergic agonist clonidine (0.5 or 2.0 mg/kg) at 10 or 42 days of age and showed age-characteristic, dose-dependent increases in locomotor activity, exploration, forelimb treading, and catalepsy. In Experiment 2, offspring were injected with clonidine (0.25 or 1.0 mg/kg) at 10, 14, or 18 days of age, and locomotor activity and wall-climbing were observed in a warm chamber (33°C). Catalepsy and rectal temperature were also scored. There were no significant differential effects of clonidine on any prenatal treatment group at any age for any measure of activity or rectal temperature. The results do not support the hypothesis that fetal ethanol exposure alters noradrenergic systems in behaviorally significant ways.

Flow cytometric and histological analysis of mouse thymus in fetal alcohol syndrome.

Reduced thymus size and fetal weight were seen in 18-day old fetuses of C57BL/6 female mice fed a complete liquid diet containing 25% ethanol-derived calories (EDC) from gestational day 0 to 18. Thymocytes from fetuses from the 25% EDC diet group and from pair-fed and ad-lib control diet groups were compared by flow cytometry for expression of thymocyte differentiation antigens. The proportions of L3T4-positive and Lyt-2-positive thymus cells were significantly reduced in alcohol-exposed fetuses compared to controls; however, the number of Thy-l-positive cells did not differ among any of the groups. Histologically, the thymus from 25% EDC fetal mice failed to show the delineation between cortex and medulla that was seen in the thymuses of control fetuses. These results indicate that thymus immaturity is one of the accompanying features of fetal alcohol syndrome.

Paternal alcohol exposure affects offspring behavior but not body or organ weights in mice.

Male mice consumed liquid alcohol diets containing 20, 10, or 0% ethanol-derived calories (EDC) for 56-61 days. Animals fed the 10 and 0% EDC diets were pair-fed to 20% EDC animals. A nontreated ad libitum group was included to assess the effects of pair-feeding. After treatment, males were bred to nontreated females. Litter size, birthweight, bodyweight at 21 or 55 days of age, and organ weights except for thymus were not affected by paternal alcohol ingestion. There was a dose-related decrease in activity at 20 and 24 days of age in an activity chamber and a dose-related decrease in serum testosterone levels at 55 days of age in male offspring sired by alcohol-consuming males. Offspring sired by males consuming the 20% EDC diet also required fewer trials to learn a passive avoidance task and had longer latencies to reach the choice point in a T maze.

Effects of paternal alcohol consumption in mice.

Male mice were divided into four groups, one group was given ad libitum access to a liquid alcohol diet containing 35% ethanol derived calories (EDC). A second group was pair fed an isocaloric control diet containing 17.5% EDC whereas a third was similarly treated with a 0% EDC diet for a minimum of 42 days. A fourth group served as ad libitum nontreated controls to assess the role of pair feeding. Males were then mated with nontreated females. The males consuming alcohol had an increased percentage of abnormal sperm and there was a significant effect of paternal alcohol exposure on implantation sites, but no effect on pre- or postnatal mortality or fetal weight. These results suggest that paternal alcohol consumption adversely affects sperm production but does not affect development of offspring in mice.

Effect of prenatal alcohol exposure on consumption of alcohol and alcohol-induced sleep time in mice

An animal model was used to examine the effect of maternal alcohol administration on behaviors in the offspring which might predispose to alcoholism. Pregnant C3H mice were administered a liquid diet containing 28% ethanol-derived calories (EDC) from Gestation-Day 8 until parturition. Control animals were either pair-fed an isocaloric 0% EDC diet or received standard lab chow and water throughout pregnancy. Offspring were tested for sleep time following a challenge dose of 3.5 or 4.5 g/kg ethanol at 25 or 110 days of age or for consumption of 10% w/v ethanol in a two-bottle choice situation. The results demonstrated that prenatal exposure to alcohol did not affect alcohol-induced sleep time at either testing age or dose and that waking blood alcohol levels were similar across groups. Voluntary alcohol consumption, however, was higher in mice exposed to alcohol in utero during the initial week of testing but intake decreased to near control levels by the third week. Whether other alcohol-related behaviors are altered by prenatal alcohol exposure remains to be examined.