Homeostasis changes induced by the action of ethanol on the materno-fetal complex in rats. V. Late fetal effects of acute intoxication during the preimplantation period

The late fetal effect of ethanol administered during the preimplantation period (in acute experiment) was investigated. Ethanol was injected i.v. (33.16% v/v in a.d., 4.80 ml/kg b.w.) to pregnant female rats on day 2 and 4 of pregnancy. Some effects concerning biochemical and morphological homeostasis on at-term fetuses (day 20 of pregnancy) were studied. Data obtained were compared with those of the control group. Biochemical investigation performed on hepatic DNA and on some serum metabolites revealed the following statistically non-significant changes: the increase of fetal hepatic DNA; the increase of total protein determined from pooled fetal serum of the whole litters; hypoalbuminemy and hyperglobulinemy; hypo-alpha 1-globulinemy and hyper-alpha 2-, beta-, gamma-globulinemy; the increase of total lipids, decrease of cholesterol; increase of uric acid and urea. In the amniotic fluid the following statistically non-significant values were found: increase of proteins, lipids, uric acid and urea content and decrease of cholesterol. Ponderal somatometry evidenced a statistically significant decrease of fetal and placental wet weight. The changes found show that--in our experimental conditions--the i.v. administration of ethanol during the preimplantation period does not significantly influence the late, fetal biochemical values and induces a significant lowering of fetal and placental wet weight and a significant increase of late fetal mortality.     

Homeostasis changes induced by the action of ethanol on the materno-fetal complex in rats. IV. Late maternal effects following acute intoxication during the preimplantation period

The possible late maternal effect on biochemical homeostasis of acute administration of ethanol during the preimplantation period was studied on pregnant albino female rats (Wistar strain). Females were injected i.v. with a 33.16% (v/v) solution of ethanol (4.80 ml/kg body weight) on day 2 and 4 of pregnancy, the animals being sacrificed on day 20. The effects on hepatic DNA biosynthesis and on some serum metabolites (proteins, lipids and some non-protein nitrogen compounds) were determined in the treated and in an untreated control group. In the treated group a non-significant increase of hepatic DNA concentration was found. Homeostasis changes of serum proteins involved: a slight increase of total serum protein content, hypoalbuminemia and hyperglobulinemia (non-significant values). Lipid metabolites showed also non-significant changes: increase of total lipids and decrease of cholesterol. Uric acid and urea (non-protein metabolites) concentration was increased. This increase, in spite of its significance level, must be taken into account and may be due to the presence of dead fetuses and to the consecutive renal lesions.     

Distortions induced in DNA by cis-platinum interstrand adducts.

A 22 base pair double-stranded oligonucleotide containing a unique interstrand adduct resulting from chelation of the two guanine residues within the central sequence d(TGCT/AGCA) by a cis-platinum residue has been studied by means of gel electrophoresis, chemical probes, and molecular mechanics. The anomalously slow electrophoretic mobility of the multimers of the platinated and ligated oligomers suggests that the platinated oligonucleotide is bent. The two cytosine residues (complementary to the platinated guanines) are hyperreactive to hydroxylamine, indicating a large exposure of the two bases to the solvent. The adduct does not induce a local denaturation within the flanking sequences since the adenine residues are not reactive with diethyl pyrocarbonate. This is confirmed by the nonreactivity of the complementary T residues with osmium tetraoxide. These results and the molecular mechanics modeling suggest that the interstrand adduct bends the double helix by approximately 55 degrees toward the major groove, that the double helix conserves its average twist angle, and that the distortion induced by the adduct is localized at the platinated sequence d(GC/CG).     

The development of bone changes induced in rats by recombinant human granulocyte colony-stimulating factor is suppressed by bisphosphonate.

We have previously demonstrated that high doses of recombinant human granulocyte colony-stimulating factor (rhG-CSF) induce bone changes characterized by osteoclastic bone resorption and osteogenesis due to intramembranous ossification in rats. In this communication we examined the effects of a pretreatment with 3-amino-1-hydroxypropylidene-1,1-bisphosphonate (AHPrBP), which is a powerful inhibitor of osteoclastic bone resorption, on bone changes induced by rhG-CSF in order to investigate the relation between osteoclastic bone resorption and osteogenesis. AHPrBP (5 mg/kg/day) was subcutaneously given to 6-week-old rats for 2 days. From the following day of the final injection of AHPrBP, rats received a subcutaneous injection of rhG-CSF (1,000 micrograms/kg/day) for 14 days, and the femur and tibia were evaluated histopathologically. By the analysis of peripheral blood leukocyte counts, spleen weights and bone marrow findings, the pretreatment with AHPrBP had no effect on the activation of hematopoiesis related to the major pharmacological activity of rhG-CSF. In the rats treated with rhG-CSF alone, accelerated osteoclastic bone resorption and osteogenesis due to intramembranous ossification were observed in the trabeculae of metaphyseal spongiosa. The accelerated osteoclastic bone resorption induced by rhG-CSF was suppressed by the pharmacological activity of AHPrBP. Furthermore, the osteogenesis induced by rhG-CSF was also suppressed by AHPrBP. These results suggest that the osteogenesis induced by rhG-CSF is a sequential reaction of accelerated osteoclastic bone resorption, and moreover that the main action of rhG-CSF on bone is an acceleration of osteoclastic bone resorption.     

Thorium induced testicular changes in rats.

Daily intraperitoneal administration of thorium nitrate produced progressive morphological and biochemical alterations with the increase in thorium concentration in rat testis. The degenerative changes in the seminiferous tubules increased with the duration of treatment and after 90 days calcification occurred in about 25% of the tubules and in the connective tissue of the tunica albuginea. The activity of adenosine triphosphatase and alkaline phosphatase increased markedly as a result of thorium administration. An attempt has been made to interrelate histopathological and enzymatic changes and the metal concentration in the testicular tissue.     

The protective effects of resveratrol against changes in blood platelet thiols induced by platinum compounds.

Cisplatin (cis-diamminedichloroplatinum II, cisPt) is especially useful in the treatment of epithelial malignancies, however, the use of cisplatin is accompanied by several toxicities including haematological toxicity. Contrary to cisplatin, selenium-cisplatin conjugate ((NH(3))(2)Pt(SeO(3)); Se-Pt) has only a slight toxicity effect on blood platelet function. In the mechanism of platinum compounds action on platelets thiols are involved. The aim of the present studies was to examine in vitro how trans-resveratrol (trans-3,4',5-trihydroxystilbene) acts on the levels of platelet glutathione (GSH) and other thiol-containing compounds and how, as an antioxidant, protecs blood platelets against the oxidative stress caused by platinum compounds (cisPt and Se-Pt). To analyse the level of thiols in human blood platelets treated with platinum compounds and with resveratrol the classical technique HPLC has been used. Blood platelets isolated by differential centrifugation of human blood were incubated (30 min, 37 degrees C) with cisPt or Se-Pt at dose of 10 microg/ml that inhibits platelet function and with resveratrol (25 microg/ml). The obtained results indicate that platinum compounds caused in platelets a decrease of both, reduced glutathione (GSH) and free thiols of cysteine (CSH) and cysteinylglycine (CGSH). The pool of these compounds in unreduced form was increased. Platinum compounds caused the reduction of platelet protein thiols. Resveratrol (after 30 min action) at the concentration of 25 microg/ml partly reduced the platinum compounds induced decrease of platelet thiols, particularly thiols in acid-soluble fraction.     

Analysis of mutations induced by alkylating compounds in Drosophila sperm. I. Character of visible changes induced by N-nitrosoethylurea and ethylmethanesulfonate in a succession of Drosophila generations]

Newly appearing hereditary traits were observed in several generations of Drosophila melanogaster: changes in eye and body colour, wing shape, number and shape of chaetae. The results obtained may be interpreted as the evidence for the realization of potential changes induced by NEU and EMS into visible mutations during 1-4th generations. X-irradiation (at a dose of 1000 r) of males from the first generation did not affect the realization of long-living potential changes induced by NEU.     

The effect of ethanol upon early development in mice and rats. I. In vivo effect upon preimplantation and early postimplantation stages

The preimplantation and early postimplantation effect of chronic alcohol consumption (at least a month before mating and during pregnancy until killing) and of acute ethanol intoxication during the preimplantation period (i.v. infection of ethanol) was studied on albino rats (Wistar) and albino mice (RAP). The main results were as follows: Chronic alcoholization. Rats: significant retardation of preimplantation development and in early postimplantation stages; a tendency of lowering of the mean litter size. Mice: significant increase of the number of preimplantation pathological forms; a tendency of lowering of the mean litter size. Pathological changes show, both in rats and mice, an obvious "litter effect". Acute ethanol intoxication. Rats: significant retardation in some litters, normal or even advanced development in others. This effect differs from the previously reported effect of acute ethanol intoxication during early postimplantation stages. The results obtained attest the prenatal noxious effect of chronic ethanol consumption in both species used and of acute ethanol intoxication during preimplantation development upon early postimplantation development in rats. Within the limits of extrapolation possibilities, they represent a risk signal for other species (including human).     

Nitrate induced changes in sensoro-motor development and learning behaviour in rats

Development of sensoro-motor functions and adult learning behaviour was studied in rats subjected to nitrate exposure. Pregnant and lactating dams and their offspring were supplied with drinking water containing nitrate, 1.12 or 2.24 mM KNO3, and compared with nitrate-free controls. Postnatal maturation of reflexes, that of sensory and somatic parameters and motor activity, furthermore, the acquisition of one-way avoidance and rewarded discriminative learning behaviour in adulthood were examined. Reflexes (righting, cliff-avoidance) and hearing startle reaction maturated earlier in nitrate treated groups. No difference was found in olfactory homing behaviour, in the time of eye opening and in body weight growth. Open-field motor activity was higher at days 5, 7, 10 after birth, but hypoactivity ensued after day 20. A marked learning deficit was observed both in punished and in rewarded learning paradigms. The results indicate a nitrate induced deviation in behavioural development, and an impairment in learning behaviour, particularly of discriminative type.