An immunohistochemical study of adenohypophyseal cells containing follicle-stimulating hormone and luteinizing hormone during the phase of selective follicle-stimulating hormone release in postnatal female rats

Summary Serum concentration of follicle-stimulating hormone (FSH) in the juvenile female rat increases independently from that of luteinizing hormone (LH). The objective of this study was to determine whether this increase in serum FSH is accompanied by a proliferation of FSH-cells greater than the proliferation of LH-cells. Thus, we measured circulating FSH and LH in female rats on days 3, 10, 13, 17, and 20, calculated the percentages of adenohypophyseal cells that contained FSH or LH on days 3, 10, and 20, and determined whether cells containing only FSH existed on day 10. Serum FSH concentrations on days 10 and 13 were significantly greater than those on days 3, 17, or 20. No differences existed in serum LH concentrations. Cells containing FSH or LH were distributed throughout the entire adenohypophyses of 3, 10, and 20-day-old females. Clusters of these cells were observed in the ventral regions of adenohypophyses of 3-day-old females. The percentages of adenohypophyseal cells containing FSH increased significantly from 9% in 3-day-old rats to 17% in 10-day-old rats and then decreased to 14% in 20-day-old animals. At all ages the percentages of adenohypophyseal cells containing FSH were similar to the percentages of cells containing LH. At 10 days of age, all cells containing FSH also contained LH and all cells containing LH also contained FSH. These data suggest that the increase in serum FSH in the juvenile female rat is associated with an increase in the percentage of adenohypophyseal cells containing FSH and that at this time all cells containing FSH also contain LH.     

Biological evaluation of various spacecraft cabin atmospheres,I

The physiological consequences of exposure to several possible spacecraft atmospheres were evaluated. Each atmosphere contained oxygen at a partial pressure of 180 mm Hg. Rabbits and rats were exposed at 1 atm abs. for one week each to atmospheres containing nitrogen, helium, argon or neon; and to pure oxygen at 200 mm Hg. In addition rats were exposed at a total pressure of 474 mm Hg to atmospheres containing nitrogen, helium or neon.Metabolic rates were increased in animals exposed to helium-oxygen at sea level, and reduced in those exposed to the low pressure, pure oxygen environment. Rates during sea-level exposures to argon and neon, and during the altitude exposures, did not differ appreciably from results obtained in air at sea level. Rabbits sustained a significant loss of hemoglobin (9%) and red blood cells during their exposure to helium-oxygen.These responses are consistent with the thermal characteristics of the several gaseous environments. A good correlation was found to exist between the calculated relative convective heat transfer in the various atmospheres and the observed metabolic rates. The possibility of an effect of helium at the molecular level has not been ruled out completely.After saturation with the inert gases studied, rats decompressed to 100 mm Hg showed the most severe symptoms of decompression sickness; nitrogen produced less damage; animals exposed to helium or neon were free of serious symptoms.The data provide the first experimental support for several theoretical advantages of neon for use in space cabin atmospheres.This work was supported by the USAF School of Aerospace Medicine under contract AF 41 (609)-2711, to Union Carbide Corporation, Tonawanda, N.Y. These experiments were conducted in 1965–1966 as a preliminary screening whose intent was to compare a wide variety of environmental and physiological parameters in a restricted number of subjects; the data should be interpreted with these limitations in mind.     

Investigation of extracellular L-citrulline concentration in the striatum during alcohol withdrawal in rats

In this study, changes in striatal extracellular L-citrulline concentrations were investigated hourly for 5 h following alcohol withdrawal in chronic alcohol feeding Wistar rats. Alcohol (7.2% ethyl alcohol, v/v) was given to rats as modified liquid diet for 20 days. Signs of alcohol withdrawal appeared from the 1st h of alcohol withdrawal and the total alcohol withdrawal scores remained higher during the course of experiments. The mean of basal levels of L-citrulline in the microdialysis samples collected in conscious rat model from the striatum of control and alcoholized rats were found to be 1.28 ± 0.48 M and 0.35 ± 0.08 M, respectively. L-citrulline levels in the striatum of alcoholized rats increased by 4 folds significantly within 1 h following alcohol withdrawal. The increased striatal L-citrulline concentration was blocked by N^G-nitro-L-arginine methyl ester (L-NAME; 60 mg/kg), a nitric oxide synthase inhibitor, pretreatment. Our results indicate an increased L-citrulline level in the rat striatum during early alcohol withdrawal and this situation may be related to an increased nitric oxide production.     

Effect of glucocorticoids on alpha-fetoprotein synthesis and liver structure in mice acutely poisoned with carbon tetrachloride]

Experiments on male C57BL mice demonstrated that the CCl4 inhalation for 15 minutes in a concentration of 0.05 ml to 4 l of air caused deep dystrophic lesions of the liver with the signs of necrobiosis. The signs of regeneration and cellular infiltration appeared almost in 24 hours accompanied by the secretion of alpha-fetoprotein into the blood. The foci of necrobiosis disappeared completely in 4 days, being replaced by the lymphoid cells with large hepatocytes in the circumference. In the animals given hydrocortisone (20 mg/kg) or dexamethasone (2 mg/kg) for therapeutic or prophylactic purpose the regenerative processes were decreased, and the number of animals producing alpha-fetoproteins fell from 92 to 60--65%.     

Fatty acid composition of rat liver mitochondrial phospholipids during ethanol inhalation

Male Sprague-Dawley rats were exposed to increasing concentrations (15-22 mg/l) of ethanol vapor over a 4-day period. Phospholipids were analyzed in liver mitochondria isolated from ethanol-treated and pair-weighted control animals. After a 2-day inhalation period, the proportion of monoenoic acids in total phospholipids increased, whereas that of arachidonic acid decreased. These changes were more striking in phosphatidylcholine (PC) than in phosphatidylethanolamine (PE). The decrease in 20:4 may be related to increased lipid peroxidation. After a 4-day inhalation period, quite different changes in phospholipid fatty acids were found. They consisted in a trend towards a more unsaturated system, the proportion of 20:4 being increased in PC and that of 22:6 in PE. This increase in polyunsaturated acids might be related to a direct ethanol effect on lipid structure and/or metabolism that would be linked to the high blood alcohol level present at this stage of ethanol intoxication.     

Possible correlations between the level of alcoholic motivation and the electrophysiological structure of sleep in the rat

Using modified Porsolt's method, the electrophysiological sleep pattern was studied in normal conditions and after a single intraperitoneal ethanol injection to noninbred male albino rats divided into 2 groups ("high activity" and "low activity" rats). Voluntary alcohol intake in these rats was measured during free choice between 10% ethanol and water for 20 days. "Low activity" rats were characterized by a statistically significant 3.4-fold higher level of ethanol consumption and 2.7-fold longer REM-sleep stage, as compared to "high activity" animals. In "low activity" animals ethanol (1 g/k, 10% solution, i. p.) inhibits and in "high activity" rats it increases REM-sleep stage, thus removing differences in the sleep pattern in the two groups of rats. The data obtained suggest a possible role of REM-sleep in the development of alcohol motivation.     

Effect of alcohol on microsomal cortisol 4en-5 alpha-reductase in the liver of rats fed on a standard or low protein diet

Alcohol feeding (40% of total calories over a period of 9 days) increases microsomal cortisol-5 alpha-reductase activity in rat liver distinctly. It is assumed that this is an adaptive response to an increased release of cortisol caused by alcohol administration. Cortisol-5 alpha-reductase activity is decreased to one third of control values in rats fed an isocaloric, low protein diet. The response to alcohol feeding is susta ined in these animals. Phenobarbital treatment (80 mg/kg x day) stimulates 5 alpha-reduction of cortisol per g of microsomes almost twofold. The activity calculated per total liver increases 4-fold. Alcohol administration has no additional effect on cortisol-5alpha-reductase in phenobarbital-treated rats.     

Induction of a putative metallothionein gene in the blood cockle, Anadara granosa, exposed to cadmium

The relationship between a putative metallothionein gene (MT) and exposure to cadmium (Cd) in blood cockles (Anadara granosa) is reported. In a 96-h dose-response experiment, mortality of cockles was found to proportionately increase in the range of 0.2-5.0 mg/l Cd with a calculated LC(50) of 2.94 mg/l. Exposure to 0.25 mg/l Cd for 16 days caused significant increases (P<0.05) in Cd concentrations in whole tissues, gills and hepatopancreas, and the accumulation of Cd in these tissues increased with the duration of exposure. Two cDNA libraries constructed using the hepatopancreas from control and Cd-treated cockles gave titres of 5.62 x 10(5) and 1.94 x 10(5) pfu/microg vector, respectively. A putative MT gene, AnaMT, of 510 nucleotides in length, was isolated from the treated cDNA library using a heterologous probe MT20 from the blue mussel, Mytilus edulis. Northern analyses using AnaMT as a probe indicated low expression of the MT mRNA in control animals. In cockles treated with 0.25 mg/l Cd for 4 days, MT mRNA level increased to approximately 168%, but declined to 108% at day 8. After 12 and 16 days of Cd treatment, expression of the MT gene was 138% and 187%, respectively, compared to the controls. These observations suggest that induction of the MT gene by a sublethal dose of Cd is rapid, occurring within 4 days of treatment.     

Lack of lead effects on fetal development and offspring learning when combined with alcohol in the Long-Evans rat

Two studies were conducted to evaluate the interactive effects of alcohol and lead during pregnancy in rats. Our purpose was to see if lead, as lead acetate, would influence the alcohol effect already known to exist. In the first study, pregnant Long-Evans rats received lead (as lead acetate), alcohol (20% w/v), or lead plus alcohol once a day on gestation days (GD) 10-20. On GD 20, when animals were sacrificed, mean blood alcohol levels were consistently higher for the lead-plus-alcohol-dosed groups compared to alcohol alone, but these two groups did not differ in maternal weight gain, percent resorptions, litter size, or fetal weight. Mean blood lead levels were not consistently higher in the lead-plus-alcohol groups compared to lead only, but the lead-plus-alcohol groups differed significantly from the lead-only groups at higher doses in the previously mentioned parameters. The lead-only groups did not differ from vehicle controls in any parameter in spite of blood lead levels as high as 300 micrograms/dl. In the second experiment, animals given a combination of alcohol and lead did not differ in activity, passive avoidance, or active avoidance learning compared to animals given alcohol or lead alone. Animals given lead only or the combination of lead plus alcohol had longer first trial latencies in the passive avoidance test. The data indicate that neither lead nor alcohol attenuates or potentiates each other's effects on reproduction or learning behavior in the Long-Evans rat even at high blood lead levels.     

Phenol utilisation by fungi isolated from activated sludge

The paper presents the efficiency of phenol removal (concentrations from 500 to 2000 mg/l) by fungi isolated from activated sludge purifying wastewater with high phenol concentration. Five fungal strains were isolated and identified. All isolated strains appeared to be Moniliales from the class of Fungi Imperfecti (Candida sp., Monosporium sp., Trichosporon sp.) Stationary cultures of the individual strains and their mixtures were maintained in Czapek medium containing phenol in concentration from 500 to 2000 mg/l. All isolated strains (except one) were capable of utilising phenol up to a concentration of 1500 mg/l. Depending on investigated strain, phenol in concentration of 500 mg/l was decomposed during 4-25 days, 750 mg/l during 4-14 days. After 20 days, a phenol decline of 1000 mg/l was observed. After 16 days, the phenol decline was 1500 mg/l. Higher phenol concentrations (1500 mg/l) were utilised only by a mixture of two strains. The investigated fungal strains showed good efficiency of phenol removal from high phenol concentration in wastewater and they may be proposed for use in the process of purifying wastewater of this type.     

Effect of central adrenergic alpha receptor blockader IEM-611 on the activity of alcohol and aldehyde dehydrogenase in rat liver

The effect of IEM-611 (30 mg/kg) on alcohol consumption in rats under the conditions of voluntary choice between water and 15% ethanol was studied as that on alcohol dehydrogenase (ADH) in postmitochondrial supernatant and in NAD-dependent aldehyde dehydrogenases (A1DH) in liver mitochondria. Administration of IEM-611 during 6 or 12 days reduces ethanol consumption by 29 and 30%, respectively, activates ADH and appreciably decreases overall activity of NAD-dependent A1DH. At the same time the ADH/A1DH ratio increases. Activation of ADH and A1DH and the decreased ADH/A1DH ratio were disclosed in alcohol preferring rats as compared to water preferring animals. IEM-611 shifts enzymatic activity of ethanol metabolism towards the level characteristic for water preferring rats. It is suggested that variation of the ADH/A1DH ratio is one of the mechanisms responsible for the decreased ethanol consumption in rats.     

Regulation of gluconeogenesis during exposure of young rats to hypoxic conditions

1. Two-day-old rats were exposed at constant temperature to atmospheres containing air and nitrogen with the air content varied in steps from 100 to 0%. By using this system of graded hypoxia a comparison was made between rates of gluconeogenesis from lactate, serine and aspartate in the whole animal and the concentrations of several liver metabolites. 2. Gluconeogenesis, expressed as the percentage incorporation of labelled isotope into glucose plus glycogen, proceeds linearly for 30min when the animals are incubated in a normal air atmosphere, but is completely suppressed if the atmosphere is 100% nitrogen. 3. Preincubation of animals for between 5 and 30min under an atmosphere containing 19% air results in the attainment of a new steady state with respect to gluconeogenesis and hepatic concentrations of ATP, ADP, AMP, lactate, pyruvate, beta-hydroxybutyrate and acetoacetate. 4. When lactate (100mumol), aspartate (20mumol) or serine (20mumol) was injected, it was shown that the more severe the hypoxia the greater the depression of gluconeogenesis. Under conditions when gluconeogenesis was markedly inhibited there were no changes in the degree of phosphorylation of hepatic adenine nucleotides, but free [NAD(+)]/[NADH] ratios fell in both cytosol and mitochondrial compartments of the liver cell. 5. Measurements of total liver NAD(+) and NADH showed that the concentrations of these nucleotide coenzymes changed less with anoxia, in comparison with the concentration ratio of free coenzymes. 6. Calculations showed that the difference in NAD(+)-NADH redox potentials between mitochondrial and cytosol compartments increased with the severity of hypoxia. 7. From the constancy of the concentrations of adenine nucleotides it is concluded that liver of hypoxic rats can conserve ATP by lowering the rate of ATP utilization for gluconeogenesis. Gluconeogenesis may be regulated in turn by the changes in mitochondrial and cytosol redox state.     

Preferential alcoholic embryopathy: effects of liquid diets.

Our initial report of a preferential expression of experimental alcoholic embryopathy affecting the male offspring contiguous in utero to male siblings of Long-Evans rats was based on gavage administration of alcohol to pregnant rats without regard to isocaloric, pair-fed exposure paradigms. In this study, pregnant Long-Evans rats were given 35% ethanol-derived calories (EDC) in one of two different liquid diets: 1) a liquid alcohol diet based on Sustacal, a flavored liquid food formulated for human nutritional standards; and 2) a high-protein liquid rodent diet devised by Lieber and DeCarli (L&D). The diets were administered from day 6 to 15 of gestation. Pregnant rats were pair-fed liquid diets containing 0% EDC, but isocalorically balanced to 35% EDC with either sucrose (Sustacal) or maltose-dextrin (L&D). A fifth group of pregnant rats was given access ad libitum to standard certified laboratory rodent diet and served as free-fed controls. On day 20 of gestation, all pregnant rats were euthanized and the products of conception examined by standard teratologic techniques. Pregnant animals fed Sustacal-based diets consistently consumed fewer calories per kilogram body weight per day from day 6 to 15 of gestation (i.e., they were significantly calorie-deprived during pregnancy) as compared with the standard laboratory-diet-fed controls or those consuming L&D diets. Body weights of rats consuming Sustacal diets (both 0 and 35% EDC) were significantly lower throughout gestation when compared with all other groups. Higher (> 150 mg/dl) blood alcohol levels were attained by rats consuming 35% EDC in Sustacal diet as compared with L&D (100 mg/dl) diets.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hepatic redox state: attenuation of the acute effects of ethanol induced by chronic ethanol consumption

The effects of an acute dose of a diet containing ethanol (3g/kg) on hepatic redox state was compared in rats fed ethanol for 25 days and in their littermates given isocaloric carbohydrate. In both groups, cytoplasmic and mitochondrial redox states of pyridine nucleotides shifted to a more reduced level, but the changes were much less extensive in rats chronically fed ethanol. This metabolic adaption may reflect the oxidation of ethanol by a pathway not involving alcohol dehydrogenase, such as the microsomal ethanol osidizing system, which increases in activity after chronic ethanol ingestion.     

Progesterone mediated increase in monoamine stores and the regulation of enzymes of biosynthesis and metabolism in the adrenal gland during late pregnancy in the rat.

The influence of repeated injections of progesterone to pregnant rats upon monoamine storage and regulation of enzymes phenylethanolamine-N-methyltransferase (PNMT), monoamine oxidase (MAO) and catechol-O-methyltransferase (COMT) was studied. All the pregnant females received progesterone (4 mg/100 g body weight) on 19, 20 and 21 days post-coitum but one group was killed at 21 days of pregnancy and the other one at 0 h parturition. Adrenal epinephrine demonstrated highly significant increase in progesterone treated rats. At the same time norepinephrine content declined significantly from the control value. The activity of enzyme PNMT also showed marked increase in the adrenals of progesterone treated females. Activity of enzyme MAO showed a slight decline after progesterone treatment to pregnant rats. Enzyme COMT in progesterone treateed animals showed decline at 0 h parturition but at 21 days post-coitum it was significantly higher from non-injected females. All the increases and decreases in monoamines and the three enzymes were significant when the results were expressed per adrenal gland or per gram of adrenal. The results suggest that exogenous progesterone administration during late pregnancy increases epinephrine stores by declining monoamine metabolism by MAO and COMT and increasing their synthesis by PNMT which is responsible for N-methylation of norepinephrine to epinephrine.     

Species Heterogeneity Between Gerbils and Rats: Quinolinate Production by Microglia and Astrocytes and Accumulations in Response to Ischemic Brain Injury and Systemic Immune Activation

Abstract: Quinolinic acid is an excitotoxic kynurenine pathway metabolite, the concentration of which increases in human brain during immune activation. The present study compared quinolinate responses to systemic and brain immune activation in gerbils and rats. Global cerebral ischemia in gerbils, but not rats, increased hippocampus indoleamine-2,3-dioxygenase activity and quinolinate levels 4 days postinjury. In a rat focal ischemia model, small increases in quinolinate concentrations occurred in infarcted regions on days 1, 3, and 7, although concentrations remained below serum values. In gerbils, systemic immune activation by an intraperitoneal injection of endotoxin (1 mg/kg of body weight) increased quinolinate levels in brain, blood, lung, liver, and spleen, with proportional increases in lung indoleamine-2,3-dioxygenase activity at 24 h postinjection. In rats, however, no significant quinolinate content changes occurred, whereas lung indoleamine-2,3-dioxygenase activity increased slightly. Gerbil, but not rat, brain microglia and peritoneal monocytes produced large quantities of [¹³C₆]-quinolinate from l -[¹³C₆]tryptophan. Gerbil astrocytes produced relatively small quantities of quinolinate, whereas rat astrocytes produced no detectable amounts. These results demonstrate that the limited capacity of rats to replicate elevations in brain and blood quinolinic acid levels in response to immune activation is attributable to blunted increases in local indoleamine-2,3-dioxygenase activity and a low capacity of microglia, astrocytes, and macrophages to convert l -tryptophan to quinolinate.     

Effect of manganese on the development of glial cells cultured from prenatally alcohol exposed rats

Maternal alcohol abuse is known to produce retardation in brain maturation and brain functions. Using cultured glial cells as a model system to study these effects of alcohol we found an alcohol antagonizing property for manganese (Mn). Mn was added to the alcohol diet (MnCl₂ 25 mg/l of 20% v/v ethanol) of pregnant rats. Glial cells were cultured during 4 weeks from cortical brain cells of pups born to these mothers. Several biochemical parameters were examined: protein levels, enzymatic markers of glial cell maturation (enolase and glutamine synthetase), superoxide dismutase a scavenger of free radicals produced during alcohol degradation. The results were compared to appropriate controls. A beneficent effect of Mn was observed for the pups weight which was no more significantly different from the control values. Protein levels, enolase and glutamine synthetase activities were increased mainly during the proliferative period when Mn was added to the alcohol diet compared to the only alcohol treated animals. This Mn effect was not found for superoxide dismutase in cultured glial cells but exists in the total brain of the 2 week-old offspring. In the total 2 and 4 week-old brain the alcohol induced decrease of enolase and glutamine synthetase was also antagonized by the Mn suplementation. Our data suggest that Mn may act as a factor overcoming at least partially some aspects of alcohol induced retardation of nerve cell development.     

Repeated alcohol administration during adolescence causes changes in the mesolimbic dopaminergic and glutamatergic systems and promotes alcohol intake in the adult rat

Adolescence is a developmental period which the risk of drug and alcohol abuse increases. Since mesolimbic dopaminergic system undergoes developmental changes during adolescence, and this system is involved in rewarding effects of drugs of abuse, we addressed the hypothesis that ethanol exposure during juvenile/adolescent period over-activates mesolimbic dopaminergic system inducing adaptations which can trigger long-term enduring behavioural effects of alcohol abuse. We treated juvenile/adolescent or adult rats with ethanol (3 g/kg) for two-consecutive days at 48-h intervals over 14-day period. Here we show that intermittent ethanol treatment during the juvenile/adolescence period alters subsequent ethanol intake. In vivo microdialysis demonstrates that ethanol elicits a similar prolonged dopamine response in the nucleus accumbens of both adolescent and adult animals pre-treated with multiple doses of ethanol, although the basal dopamine levels were higher in ethanol-treated adolescents than in adult-treated animals. Repeated ethanol administration also down-regulates the expression of DRD2 and NMDAR2B phosphorylation in prefrontal cortex of adolescent animals, but not of adult rats. Finally, ethanol treatment during adolescence changes the acetylation of histones H3 and H4 in frontal cortex, nucleus accumbens and striatum, suggesting chromatin remodelling changes. In summary, our findings demonstrate the sensitivity of adolescent brain to ethanol effects on dopaminergic and glutamatergic neurotransmission, and suggest that abnormal plasticity in reward-related processes and epigenetic mechanisms could contribute to the vulnerability of adolescents to alcohol addiction.     

Heat acclimation causes some alterations in serum enzymes activities in streptozotocin-diabetic rats

The present study investigated the effect of long-term heat acclimation and experimental diabetes on serum activity of transaminases (AST, ALT), ALP, LDH and elastase complex, as well as blood glucose and HbA1c level in Wistar rats. The heat acclimation model was established with the artificial heat chamber (35±1 °C and 30-40% humidity) for a period of 28 days, while the control groups were held on 20±2 °C. Experimental diabetes was induced by single streptozotocine (STZ) injection (55 mg/kg bodyweight) The changes caused by insulin treatment (2 IU/100 g body weight, 14 days, twice daily) in both thermal groups were also investigated.STZ-diabetes leads to significant increase in blood glucose and HbA1c level, AST, ALT and ALP activities in both thermal groups (normothermic and heat acclimated), decrease in LDH activity in normothermic animals and increase in heat-acclimated ones. Treatment with insulin restores the blood glucose, HbA1c and enzymes activities regardless of the previous thermal exposure.Prolonged acclimation of control animals to elevated ambient temperature resulted in significant decrease in blood glucose level, AST, ALT, ALP and LDH activities and non-significant changes in HbA1c. Compared to diabetic rats from room temperature, heat-acclimated diabetic ones have significantly higher blood glucose, AST, ALP and LDH activity, lower HbA1c concentration and no significant changes in ALT. Most of the changes observed in heat-acclimated insulin-treated diabetic rats did not significantly differ compared to those from room temperature.The overall two-way ANOVA analyses showed that diabetic state causes significant changes in the blood glucose, HbA1c, AST, ALT and ALP activity, while heat acclimation causes significant changes only in HbA1c level and AST activity. Both of the factors (diabetic state and heat acclimation) have significant common effects on AST, ALP and LDH activity.     

Effects of excess and deficiency of thyroid hormones in the body upon blood melatonin in pubertal male rats

Experiments have been carried out on Wistar line pubertal male rats in Winter. Radioimmunological method showed, that in intact animals the night peak of melatonin in blood was 994,36 +/- 195,99 pM/l, following ten-days intramuscular thyroxin injection (100 mg/100 g body mass)--2560,52 +/- 322,04 pM/1 and 20 days after, following bilateral thyroidectomy--117,13 +/- 16,35 pM/l, that totals 257,5% and 11,8%, respectively. Thus, the night peak of melatonin depends upon thyroid hormones concentration in blood.