Very high daily intraperitoneal doses of carbonyl compounds affect the morphology, but not the exchange characteristics, of rat peritoneum.

Glucose degradation products (GDP) are carbonyl compounds, that are formed by heat sterilization of conventional peritoneal dialysis (PD) fluids. Carbonyl compounds are known to be toxic in vitro and potentially toxic also in vivo. The aim of this study was to evaluate the effects of daily, short-term exposure of the peritoneum to very high concentrations of GDP in vivo on peritoneal transport parameters and on peritoneal morphology in a well-established rat model of PD. Rats were exposed to three daily intraperitoneal (IP) injections (10 ml) for 9 days of a largely neutral (pH 7.2) PD fluid containing 1.5% glucose and sterilized by filtration, with (n = 8) or without (n = 8) the presence of different carbonyl compounds in concentrations 100 times higher than those reported in commercial PD fluids. Seven rats, not subjected to any exposure, served as controls. After the exposure, the rats were subjected to acute PD in 4-hour dwells. Twenty milliliters of 4% glucose dialysis fluid were instilled into the rat peritoneal cavity. Blood and dialysate samples were taken during the dwell for measurements of dialysate sodium, and for assessments of the mass transfer area coefficient (PS) for glucose and 51Cr-EDTA and of transperitoneal clearance (Cl) or radiolabelled albumin (RISA). At the end of the dwell, parts of the liver, diaphragm and peritoneum were removed for measurements of tissue cell density and thickness of the submesothelial peritoneal tissue. The exposure of the peritoneum to very high doses of carbonyl compounds did not affect the peritoneal transport of fluid and small solutes significantly, but seemed to slightly reduce lymph flow and albumin clearance out of the peritoneal cavity. Assessed after a hypertonic dwell, and compared to the situation in nontreated rats after the same kind of dwell, there was a significant thinning of the submesothelial tissue, but no difference in tissue cell density. It is concluded that short-term exposure of the peritoneum in vivo to very high doses of GDP resulted in almost no signs of acute toxicity.

Glucose degradation products in peritoneal dialysis fluids may have both local and systemic effects: a study of residual fluid and mesothelial cells.

OBJECTIVE: When peritoneal dialysis (PD) fluids are heat sterilized, glucose is degraded to carbonyl compounds. These compounds are known to interfere with many cellular functions and to promote the formation of advanced glycation end-products. However, little is known about what actually happens with glucose degradation products (GDPs) after infusion into the peritoneal cavity. The aim of the present study was to investigate possible targets for GDPs in the peritoneal cavity. DESIGN: In vitro reactions between residual fluid and GDPs were studied by incubating unused PD fluid with overnight dialysate. Confluent monolayer cultures of human mesothelial cells were used as a model to study the reactions of GDPs with the cells lining the peritoneal cavity. METHODS: Samples were analyzed, using high pressure liquid chromatography, for the presence of formaldehyde, acetaldehyde, 5-hydroxymethyl-2-furaldehyde (5-HMF), methylglyoxal, and 3-deoxyglucosone (3-DG). Cytotoxicity was determined as inhibition of proliferation of cultured fibroblasts. RESULTS: None of the analyzed GDPs reacted with overnight dialysate. Formaldehyde and methylglyoxal, in contrast to 3-DG and 5-HMF, reacted with the cultured mesothelial cells. CONCLUSIONS: Low molecular weight carbonyls such as formaldehyde and methylglyoxal most probably react with the mesothelial cells lining the peritoneal cavity, and could be responsible for the disappearance of these cells during long-term treatment. 3-Deoxyglucosone showed remarkably low reactivity and was most probably transported within the patient.

Biological significance of reducing glucose degradation products in peritoneal dialysis fluids.

UNLABELLED: Carbohydrates are not stable when exposed to energy; they degrade into new molecules. In peritoneal dialysis (PD) fluids, degradation of glucose occurs during the heat sterilization procedure. The biological consequences of this degradation are side effects such as impaired proliferation and impaired host defense mechanisms, demonstrated in vitro for a great variety of cells. Several highly toxic compounds--such as formaldehyde and 3-deoxyglucosone--have been identified in PD fluids. Carbonyl compounds, apart from being cytotoxic, are also well-known promoters of irreversible advanced glycation end-products (AGEs), which might participate in the long-term remodeling of the peritoneal membrane. Various approaches can be used to reduce the formation of glucose degradation products (GDPs) during heat sterilization. Some examples are shortening the sterilization time, lowering the pH, removing catalyzing substances, and increasing glucose concentration. The latter three factors are employed in the multi-compartment bag with a separate chamber containing pure glucose at high concentration and low pH. Gambrosol trio, a PD fluid produced in this way, shows reduced cytotoxicity, normalized host defense reactions, less AGE formation, and reduced concentrations of formaldehyde and 3-deoxyglucosone. Moreover, in the clinical situation, the fluid turns out to be more biocompatible for the patient, causing less mesothelial cell damage, which in the long term could lead to a more intact peritoneal membrane. CONCLUSION: Glucose degradation products in heat-sterilized fluids for peritoneal dialysis are cytotoxic, promote AGE formation, and cause negative side effects for the patient. Using improved and well-controlled manufacturing processes, it is possible to produce sterile PD fluids with glucose as the osmotic agent but without the negative side effects related to GDPs.

Effects of acidity, glucose degradation products, and dialysis fluid buffer choice on peritoneal solute and fluid transport in rats.

OBJECTIVE: To evaluate the effects of acidity, glucose degradation products (GDP), and different solution buffer systems on solute and fluid transport during acute peritoneal dialysis (PD) in rats. DESIGN: Dialysis fluid (16 mL) containing 2.5% glucose as the osmotic agent was instilled intraperitoneally in Wistar rats (280 g) via a thin catheter in dwells lasting 4 hours. Blood and dialysis fluid samples (25 microL) were taken for measurement of glucose, sodium, and radioactive markers. The mass transfer area coefficient (MTAC or PS) for glucose and for 51Cr-EDTA (given as an intravenous infusion) and the peritoneal clearance (Cl) of 125I albumin (RISA), as well as the clearance of RISA to plasma (Cl --> P) were assessed for a commercial, heat-sterilized, acidic PD solution (2.5% glucose, pH 5.5; Gambrosol, Gambro, Lund, Sweden), containing GDP, and for four filter-sterilized solutions containing either lactate (40 mmol/L, pH 5.5 or 7.2), bicarbonate (38 mmol/L, pH 7.2), or pyruvate (40 mmol/L, pH 7.2) as buffers and being devoid of GDP. RESULTS: The initial pH of the acidic solutions increased rapidly, and attained physiological levels within 40 minutes. The initial drop of sodium, which is expected during the first part of the dwell, was significantly more pronounced with neutral than with acidic lactate. The PS for glucose and 51Cr-EDTA were slightly, but significantly, higher with the acidic and heat-sterilized solution (Gambrosol) than with the neutral, sterile-filtered lactate-buffered solution (p < 0.01), especially early during the dwell. Such an increase may be due to initial vasodilatation, and hence, recruitment of capillaries by the combination of acidity and GDP. However, there were no significant differences with respect to small solute PS values among sterile-filtered solutions, regardless of the presence of acidity or of buffer choice. CONCLUSION: There were no major differences in fluid and solute transport among sterile-filtered PD solutions having differing buffer systems and pH. Neither were there any effects of GDP alone. However, the combination of a low pH and the presence of GDP in the PD solutions seemed to cause significant increases in peritoneal small solute transport.

Limited effects of ozone exposure during pregnancy on physical and neurobehavioral development of CD-1 mice.

Only a few studies have attempted to assess in laboratory rodents the maternal toxicity and behavioral changes in offspring caused by prenatal exposure to ozone (O3). In particular, no data are available concerning the behavioral development of mouse offspring after maternal exposure, despite the fact that increasing use is made of this species in behavioral teratology studies for reasons both of economy and in order to increase the effectiveness of subsequent higher-tier studies (e.g., of treatment-genotype interactions). In the present work, female CD-1 mice were exposed during pregnancy (Days 7-17) to different O3 concentrations (0, 0.4, 0.8, or 1.2 ppm); to avoid confounding by postnatal maternal effects, all litters were assigned shortly after birth to foster dams neither treated nor handled during pregnancy. The dams' food and water intake and body weight gain were depressed in a concentration-dependent fashion. Tolerance to these effects developed during continuing exposure; such tolerance was faster in the case of food than water intake. Several measures of reproductive performance, such as proportion of pregnancies carried to term, litter size, sex ratio, frequency of stillbirth, and neonatal mortality, failed to show differences between control and O3 animals. Postnatal body weight gain was slightly but significantly depressed in the 1.2 ppm offspring. Otherwise, the somatic development of O3 pups was indistinguishable from that of controls, save for a delay in eye opening; this effect, however, failed to show a significant concentration dependence. Negative results were obtained in a wide range of assessments concerning (i) the development of various reflexes and responses ("Fox battery") from birth to Day 18; (ii) ultrasonic emissions on Postnatal Days 3, 7, and 11; and (iii) activity, habituation, response to an unfamiliar object, and hyperactivity produced by a monoaminergic stimulant (d-amphetamine) at 60-61 days. The present data differ from those of a previous study on rats raised by their biological mothers after gestational exposure to O3 (1 and 1.5 ppm), which showed a substantial impairment in somatic and neurobehavioral development (R. Kavlock, E. Meyer, and C. T. Grabowski, 1980, Toxicol. Lett. 5, 3-9). This difference, be it due to species factors, to postnatal maternal effects, or to the time of occurrence of maximal O3 effects (e.g., on food and water intake) after the onset of exposure and before adaptation or tolerance, may provide significant cues for the understanding of O3 effects in pregnant and developing organisms.

Effects of acute and continuous ozone (O3) exposure on activity/exploration and social behavior of CD-1 mice.

This study was aimed at investigating the behavioral effects of ozone (O3) exposure in CD-1 mice. Pairs of same-sex adult male and female mice were continuously exposed for 13 days to either 0, 0.4, 0.8, or 1.2 ppm O3. The exposure apparatus consisted of a system for O3 production and delivery into four stainless-steel chambers, each equipped to contain up to 24 home cages, with continuous monitoring and recording of concentrations. Acute behavioral changes were assessed during the first hour of O3 exposure without removing animals from the chambers. The onset of exposure produced remarkable behavioral disturbances consisting of a sharp increase of several responses (rearing, sniffing, grooming, feeding, and social interactions) paralleled by a reduction of bar-holding. These changes were rapidly reversed within 1 hour, suggesting that they constituted a response to strong novel stimulation followed by habituation. Subsequently, brief sessions of videorecording of the animals' activities in freshly cleaned cages (identical to the home cages) were performed outside the chambers after 3, 7, and 10 days of exposure. These tests showed a significant concentration-dependent increase of grooming and rearing and a decrease of crossing and wall climbing. Both food and water intake showed a nonmonotonic trend over time consisting of a concentration-dependent depression (for about 3 and 7 days, respectively) followed by recovery; body weight followed a similar trend. The detailed study of various components of the animal's behavioral repertoire, showing concentration-dependent and time-dependent changes in different directions, appears to be a sensitive tool in the analysis of pollutants' effects.

Acute and chronic sulphur dioxide (SO2) exposure: an overview of its effects on humans and laboratory animals.

Sulphur dioxide (SO2) is a common air pollutant found both in indoor and outdoor environments. Studies of controlled human exposure as well as epidemiological and animal investigations have documented several short- and long-term effects of SO2 exposure on the respiratory and other systems. Exercise, duration and other exposure factors may potentiate the pollutant's effects, especially in sensitive individuals such as children and asthmatics. Early postnatal somatic and behavioural alterations have been shown after maternal SO2 exposure, during pregnancy and neonatal exposure. Such exposure should be considered as a complex toxic hazard which may interfere with the developmental processes in the offspring.

Impaired maternal behaviour and altered central serotonergic activity in the adult offspring of chronically ethanol treated dams.

Female rats were given 16% ethanol solution as the sole liquid during the entire period of gestation. At birth the offspring was removed and reared by foster dams consuming normal tap water. At adult age the female offspring showed deficiencies in their maternal behaviour; they built nests of poor quality and they displayed prolonged times for retrieving pups placed outside the nest. In the whole brains of the prenatally ethanol-exposed females a decreased serotonin synthesis was observed. The offspring of the prenatally ethanol exposed mothers did not show any signs of disturbances in physical or behavioural development.

Effects of maternal ethanol consumption on the offspring sensory-motor development, ultrasonic vocalization, audiogenic immobility reaction and brain monoamine synthesis.

Female rats were given 16% ethanol solution as the sole liquid during the entire period of gestation. At birth the offspring was removed and reared by foster dams consuming normal tap water. The development of sensory motor behaviour and emotional reactions was delayed by 1-2 days in the prenatally ethanol exposed pups as assessed by tests on body righting, acoustic startle response, air righting, rearing and ultrasonic vocalization. In the open-field test the normally occurring behavioural difference between the sexes was not found in the prenatally ethanol exposed pups. Both sexes of the ethanol exposed pups behaved like the female controls suggesting deficient masculinization of the ethanol exposed male pups during foetal age. Biochemical analysis of the brains showed a decreased synthesis of serotonin and dopamine.

Effect of diazepam, apomorphine and haloperidol on the audiogenic immobility reaction and on the open field behavior.

Weanling rats were treated with diazepam, apomorphine, and haloperidol to study the influence of the dopamine (DA) system on the audiogenic immobility reaction and open field locomotory behavior. Treatment with diazepam (0.025, 0.05, and 0.1 mg/kg) caused a dose-dependent shortening of the duration of the immobility response. Treatment with apomorphine (0.125, 0.25, and 0.50 mg/kg) shortened both the immobility reaction and the latency to leave the spot where the animal was first placed in the open field (latency for first crossing). Locomotor activity increased in a dose-dependent fashion. Both grooming and rearing showed biphasic dose response curves, with a maximum occurring at the 0.125 mg/dose for grooming and at the 0.25 mg/dose for rearing. Haloperidol (0.06 mg/kg) exerted opposite effects to those of apomorphine, but also produced increased running during the auditory stimulation (flight distance). Using the immobility reaction as an expression of fear, we concluded that activation of the DA system decreases while inhibition of the DA system increases fear. It was hypothesized that the DA system exerts an inhibitory function in the expression of fear.

Development of sexual behavior in prenatally ethanol-exposed rats.

Female rats were given 16% ethanol solution as the sole liquid during the entire period of gestation. At birth the offspring was removed and reared by foster dams consuming normal drinking water. When tested for feminine sexual behavior in adulthood, the males showed marked signs of feminization as evidenced by an increased amount of lordosis responses. No changes were seen in the masculine sexual behavior. No deviations were seen in the female estrous cycles or in onset of vaginal estrus, whereas the onset of behavioral estrus was delayed. It is suggested that prenatal ethanol exposure may lower the fetal testosterone production and thereby interfere with the normal course of sexual differentiation in the male.

The effect of medial preoptic-anterior hypothalamic lesions on testosterone plasma levels and testosterone conversion in the hypothalamus of male rats.

Male Wistar rats were submitted to bilateral high frequency lesions in the medial preoptic-anterior hypothalamic area or to sham procedure. The behavioral effect of the lesions was observed and plasma testosterone (T) and estradiol (E2) were measured by radioimmunoassay. In vitro metabolism of T was studied in the hypothalamus. Lesions produced a permanent deficit in male sexual behavior, an increase of plasma T and E2, and of hypothalamic T aromatization, and a decrease of T conversion to 5 alpha-dihydrotestosterone (DHT).

Effect of castration on hypothalamic testosterone metabolism in the male rat.

In vitro studies were performed of hypothalamic testosterone (T) metabolism 30 days after castration of adult male rats. No changes were seen in T conversion into dihydrotesterone and estrogens in the castrated rats. Plasma T levels were decreased while plasma estradiol concentrations did not differ from those of intact controls. It was suggested that the hypothalamic T metabolism probably is not androgen dependent.