A biphasic developmental pattern of circulating leptin in the male rhesus macaque (Macaca mulatta)

To help elucidate the physiological role of leptin during somatic and sexual maturation, circulating concentrations of leptin were measured in 36 male rhesus monkeys of ages ranging from 0-20 yr. The body weight of these animals showed a steady increase of approximately 1 kg/yr during the first decade of life and reached a plateau at approximately 13 yr. In contrast, serum leptin concentrations showed a biphasic developmental pattern, which was highlighted by a strong negative correlation with body weight (r = -0.74, P < 0.001) before the onset of puberty (at approximately 3.5 yr) and by a strong positive correlation afterward (r = 0.77, P < 0.001). Overall, the developmental changes in serum leptin concentrations closely mimicked the expected developmental changes in serum testosterone concentrations (r = 0.62, P < 0.001), which were highly elevated at birth, fell to basal levels during the juvenile phase of development, and gradually rose again after the initiation of puberty. However, mean serum leptin concentrations during the peripubertal period itself (3-5 yr) were significantly lower (P < 0.01) than those observed during the first year of life or those observed in fully mature adults (i.e. > 7 yr) (3.5 +/- 0.3, 1.4 +/- 0.2, and 3.3 +/- 0.6 ng/ml, respectively). These data demonstrate that the role of leptin in energy homeostasis of primates is more than a simple linear relationship, being highly dependent upon the developmental age. Furthermore, the data do not support the hypothesis that leptin plays a major role in triggering the onset of puberty in primates, although the strong correlation between serum concentrations of leptin and testosterone suggests that the secretion of these two hormones may be causally linked.     

The influence of CSF calcium and magnesium on the ventilatory response to carbon dioxide during hyperoxia

Respiratory responses to inhaled carbon dioxide were measured in anaesthetized cats during perfusion of the ventriculocisternal system with artificial cerebrospinal fluid. A study was performed to evaluate the effect of changes in the magnesium and/or calcium concentration of the CSF on the CO2 response curve which was described as VE = S (PCSF, CO2 -- B). A decrease of S was observed when the magnesium concentration of the perfusion fluid was increased; the B-value remaining the same. The reverse was true down to magnesium concentrations of 0.6 mmol-1-1. Below this concentration S remained the same or decreased; the B-value was lowered. When both the calcium and magnesium concentrations of the CSF were changed, the relation between S and these concentrations could be described as to be proportional to CCAa-CMgb. The effect of changes in the calcium concentration was much more pronounced than comparable changes of the magnesium concentration as reflected by the magnitude of the exponents a and b which were found to be -2.80 (S.D. 0.11) and -0.60 (S.D. 0.03) respectively.     

Regional Analysis of Nonmethane Volatile Organic Compounds in the Lower Troposphere of the Southeast United States

Nonmethane organic compounds (NMOCs) along with ozone as well as other trace gas and meteorological parameters were measured at eight rural sites located in the Southeast United States, as part of the Southern Oxidants Study. Fifty-four C2–C10 NMOCs were collected from 1,200–1,300 local time, once every six days from September 1992 through October 1993 and intermittently during 1994. This study was undertaken to characterize the nonmethane hydrocarbons in the rural areas of Southeast United States with respect to their concentrations, reactivities, and relative importance of natural and anthropogenic abundances of NMOCs. Though the sites are well removed from large urban source regions, the observations show a clear anthropogenic influence on the hydrocarbon levels at these rural sites. The data for the sites show similar seasonal patterns for total NMOC with summer maxima (average concentrations of 198 ppbC at the Long Creek, South Carolina, site to 47 ppbC at the Candor, North Carolina site) and fall minima (average concentrations of 73 ppbC at the Long Creek site to 31 ppbC at the Centreville, Alabama site). A secondary maximum is observed during the winter. A seasonal trend was observed in the concentrations of light molecular weight C2–C4 NMOCs (ethane, ethene, acetylene, propane, i-butane, and n-butane) with a winter maximum and a summer minimum. An analysis of changes in C2–C4 hydrocarbon ratios over a period of one year indicates that the variation is most likely due to seasonal changes in OH concentrations. A seasonal trend was also observed for the biogenically emitted NMOC, isoprene, with summer maxima (average concentrations of 37 ppbC at the Long Creek, South Carolina, site to 8.6 ppbC at the Giles County, Tennessee and Metter, Georgia, sites) and winter minima with winter seasonal values below the level of detection. Isoprene was observed to be the most dominant NMOC at most sites during the summer. The ambient concentrations of isoprene measured during the summer were found to be dependent on the ambient temperature. The monoterpenes a-pinene, b-pinene, and d-limonene also peaked during the summer with averages ranging between 3.19 ppbC (Centreville, Alabama) and 6.38 ppbC (Oak Grove, Missouri), and a background concentration of 1.25 to 1.9 ppbC for all the sites during the winter.     

Clinical application of the relaxation response in women''s health

A study was performed investigating the daily patterns of hormone release accompanying changes in fluid balance in the male rat during 48 h of dehydration. The blood volume decreased by 18%, the largest change occurring during the initial period when the rats showed an effective loss of body sodium. During the second day of dehydration, sodium retention was again seen. Plasma sodium concentrations showed a progressive increase, the total rise being 5-6%; the greatest changes were seen during the dark phases of the cycle which may be due to the nocturnal food intake. Plasma vasopressin and oxytocin concentrations were significantly elevated throughout dehydration to levels which could be reproduced by acutely increasing plasma sodium and decreasing blood volume to the same extent. The observed increases were influenced by the phase of the day-night cycle, being greatest over the dark phases of the cycle. The overall increases were greatest when dehydration commenced at the start of the dark phase. Dehydration initially led to a rise in plasma corticosterone concentrations, whilst plasma concentrations of atrial natriuretic peptide were decreased. Plasma angiotensin II concentrations rose significantly during the later period of sodium retention.     

Leaching of CuFeS2 by aqueous FeCl3, HCl,and NaCl: Effects of solution composition and limited oxidant

Batch leaching experiments were performed in which the initial amounts of chalcopyrite and ferric chloride were selected to ensure that the oxidant was significantly depleted over the course of an experiment. Solution samples were analyzed for Cu(II) and Fe(III) by visible spectrophotometry and for total copper and total iron by atomic absorption, making it possible to measure changes in the solution component concentrations as leaching progressed. For selected samples, the solution potential was also measured. In all experiments, the Cu(II) concentration passed through a maximum and, simultaneously, the Cu(I) concentration increased very sharply. An acceleration in the total rate of leaching was normally observed at the same time. Early in a leach, the solution potential was too high for the reduction of Cu(II) to Cu(I) to take place at the time of the increase in the overall leaching rate, however, the solution potential dropped sharply during a span of a few hours, reaching a value low enough that reduction of cupric ion became possible. The amount of Cu(I) present at the completion of a leach was dependent on the total chloride concentration of the system. The highest Cu(I)/Cu ratios were observed in systems with the highest chloride concentrations. The ultimate extent of CuFeS₂ leaching was dependent on the initial FeCl₃ and total chloride concentrations; the FeCl₃ was virtually completely consumed and the total chloride concentration controlled the extent to which Cu(II) was reduced by reaction with chalcopyrite.     

Day and night dysfunction in intraretinal melatonin and related indoleamines metabolism, correlated with the development of glaucoma-like disorder in an avian model

As previous studies have suggested that melatonin and serotonin may be involved in the regulation of intraocular pressure, retinal concentrations of melatonin, 5-HT, and related indoleamines measured at day and at night were studied during the development of a glaucoma-like disorder with increased intraocular pressure in the al mutant quail. Indoleamine levels were determined by HPLC with electrochemical detection in 1-month-, 3-month-, and 7-month-old al mutant and control quails. Morphology and numbers of melatonin-synthesizing and 5-HT-containing cells, labelled immunohistochemically with an anti-hydroxyindol-0-methyltransferase (HIOMT) antibody and an anti-5-HT antibody, respectively, were studied. Major findings were that: (1) no significant changes in morphology of melatonin-synthesizing cells or in the morphology and density of 5-HT-containing amacrine cells were observed during the development of glaucoma: (2) 5-HT metabolism was modified during the night at 1 month of age and during the day after 3 months; and (3) melatonin metabolism was modified during the night at 7 months and during the day after 3 months. These results demonstrate a relationship between the temporal evolution of this avian glaucoma and a dysfunction in indoleamine retinal metabolism.     

Variations of rat brain calmodulin content in dark and light phases: effect of pentylenetetrazol-induced kindling

Calmodulin (CaM) through activation of CaM-kinase II may be involved in the molecular mechanisms underlying the epileptogenic processes. Some evidence suggests that kindling responses change across the day-night cycle. In order to test if kindling stimulation modifies CaM content, we measured CaM concentrations in amygdala, hippocampus and hypothalamus obtained from control and kindled rats during light and darkness. Male Wistar rats (250-300 g), were injected i.p. with Pentylenetetrazol (PTZ) (35 mg/kg/24 h). Once chemical kindling was established, rats were sacrificed by decapitation at 10:30 a.m. and 01:30 a.m. The brains were obtained, and the amygdala, hippocampus and hypothalamus dissected. CaM content was measured in the cytosol and membrane fractions by radioimmunoassay. We found a significant increase in CaM content in cytosol and membrane fractions of both control and kindled rats during the dark phase. No significant differences in CaM concentrations were observed between control and experimental rats, whether during the light or the dark phase. The data suggest a well defined photoperiodic variation in CaM concentrations in limbic structures, despite the neuronal excitability produced by kindling. In addition, the observed CaM increases during the dark time may be related to a protective mechanism against enhanced sensitivity to seizures observed during the night.     

Time course of PCDD/PCDF/PCB concentrations in breast-feeding mothers and their infants

PCDD/PCDF/PCB concentrations were measured in samples from four mothers (at delivery and during lactation) and their infants (at birth and the end of first year of life). For two of these mothers it was the second delivery and breast-feeding period, and additional data were available from first lactation period and the first-born infant at the age of 11 to 12 months. Five of the six infants were fully breast-fed for at least 17 weeks. In four of them a distinct PCDD/PCDF/PCB accumulation was observed at the end of the first year of life: concentrations in blood fat were 1.5 to 3.6 times higher than maternal levels measured at the same time. Due to decreasing maternal body burdens during lactation, PCDD/PCDF concentrations at 11 to 12 months of life were only about half as high in the second infant as in the first one at the same age. During second pregnancy, no important change of the concentrations was observed in maternal blood.     

Cytosolic estrogen receptor concentrations in the lumbosacral spinal cord fluctuate during the estrous cycle

Estrogen responsive neurons have been anatomically identified with autoradiographic and immunohistochemical techniques and their distribution mapped in the lumbosacral spinal cord of female rats. Such neurons contain estrogen receptors (ERs). The present study was undertaken to: 1) quantify cytosolic estrogen receptor (ER) concentrations in the lumbosacral spinal cord and 2) determine if there is a relationship between cytosolic ER concentrations and fluctuations in serum estradiol (SE2) levels during the estrous cycle. Lumbosacral spinal segments were removed from intact cycling rats during the morning of proestrus, the afternoon of proestrus, and the morning of estrus, metestrus and diestrus. Trunk blood was collected at euthanasia and SE2 levels were determined using radioimmunoassay. Cytosolic ER concentrations were measured using a dextran-charcoal coated tube method. Concentrations of cytosolic ERs were low during estrus and metestrus, increased during diestrus with maximum concentrations during the afternoon of proestrus. These changes in ER concentrations paralleled SE2 levels measured in intact cycling animals; i.e., during estrus SE2 levels were low, but began to rise during metestrus, diestrus, and during the morning of proestrus with a maximum peak increase during the afternoon of proestrus. These data indicate there are fluctuations of cytosolic ER concentrations during the estrous cycle and that these changes coincide with changing SE2 concentrations suggesting that ER content is influenced by SE2.     

In vitro influence of phenylalanine on acetylcholinesterase activity and DNA

Acetylcholinesterase (AChE) is a significant component of the membrane contributing to the permeability changes during synaptic transmission and conduction. Phenylketonuria is a group of metabolic disorders in which phenylalanine (Phe) is highly elevated in blood (up to 0.1 M) resulting in mental retardation etc. AChE activity was measured spectrophotometrically after incubation with various Phe concentrations. Phe interaction with DNA was evaluated with an established HPLC method. Phe was found to inhibit AChE almost 40%, at a concentration of 5 mM, whereas a 62.5% DNA peak exclusion (molecular interaction) was observed when Phe was incubated with DNA at a concentration of 3 mM. In addition the ratio of DNA: Phe determined the potency of the observed molecular effect.     

Retinoic acids induce growth inhibition and apoptosis in adult T-cell leukemia (ATL) cell lines

In order to elucidate the mechanisms of cisplatin (cis-diamminedichloroplatinum; CDDP)-resistant tumor cells, we previously established a CDDP-resistant KB cell line (KBrc cells) from a parental KB cell line derived from epidermoid carcinoma (KB cells). The KBrc cells were resistant to 5 kinds of platinum (Pt) drugs. Intracellular Pt concentrations in KBrc cells were lower than in KB cells. Decrease of intracellular Pt concentrations was one of the CDDP-resistant mechanisms. When we measured changes of intracellular calcium ion concentration ([Ca2+]i) during exposure to high-dose CDDP, a sustained elevation of the [Ca2+]i level was observed in the KB cells. These results suggest that the mechanisms underlying CDDP resistance involve changes in calcium channels and an alteration of calcium homeostasis in the tumor cell line.     

Perioperative changes in complement associated with cardiopulmonary bypass

The total haemolytic complement (CH50), the complement components C3 and C4, the complement breakdown product C3d, alternative pathway activation and transferrin, were measured before, during and after cardiopulmonary bypass. As expected, CH50 decreased after heparinization, remained low during bypass and decreased further up to 8 h after bypass. C3 and C4 decreased significantly during bypass, continued to decrease for a further 8 h after bypass (by 35% and 40% respectively) and thereafter increased gradually up to 48 h. Although the depletions observed were suggestive of complement activation, there were no demonstrable increases in C3d, and in all patients the concentration of C3d remained within the normal range. Hence it was concluded that complement depletions of this magnitude were unlikely to result from complement activation. Non-specific changes in protein concentrations during bypass, as a result of dilution, redistribution or other unidentified factors, are more probable causes of the observed reductions. The acute phase response to surgery may be a factor in the subsequent increase in C3 and C4 which is seen 24 h after bypass. As transferrin concentrations in the plasma are known to decrease during this response the observed decrease in transferrin concentration would support this view.     

Effect of graded exercise on esophageal motility and gastroesophageal reflux in nontrained subjects

The effects of graded exercise on esophageal motility and gastroesophageal reflux were evaluated in nine nontrained subjects, using a catheter with three strain-gauge transducers connected to a solid-state datalogger and an ambulatory intraesophageal pH monitor. Subjects exercised on a stationary bike at 45%, 60%, 75%, and 90% of peak O2 uptake (VO2 max). Durations of exercise sessions and rest periods varied among subjects. Studies were performed after an overnight fast and subjects received only intravenous infusion of 5% glucose solution during the study. Plasma concentrations of gastrin, motilin, glucagon, pancreatic polypeptide (PP), and vasoactive intestinal peptide (VIP) were determined at rest and before and after each exercise session. The duration, amplitude, and frequency of esophageal contractions declined with increasing exercise intensity, and the differences were significant (P < or = 0.05) for all three variables at 90% VO2 max. The number of gastroesophageal reflux episodes and the duration of esophageal acid exposure were significantly (P < or = 0.05) increased during exercise at 90% VO2 max. Plasma regulatory peptide concentrations showed no significant changes between rest and the various exercise sessions. Thus, exercise has profound effects on esophageal contractions and gastroesophageal reflux, which are intensity dependent. These effects were not mediated by the hormones measured. The results were similar to those observed in highly trained athletes, suggesting that the effects of exercise on esophageal function are similar in trained and nontrained subjects performing at similar percentages of VO2 max, even though the absolute levels of exercise achieved in each group are different.     

Depletion of adipose tissue and peripheral nerve alpha-tocopherol in adult dogs

To assess the relationship between tissue alpha-tocopherol depletion and histopathologic or functional changes in nervous tissue, a longitudinal study of male 1-year-old beagle dogs, two fed a vitamin E-deficient diet (0.05 +/- 0.02 mg alpha-tocopherol/kg;--E dogs) and two fed a vitamin E-supplemented diet (114 +/- 14 mg alpha-tocopherol/kg; +E dogs), was carried out. Plasma and adipose tissue alpha-tocopherol concentrations, neurological examinations, and sensory and motor nerve conduction velocities were determined at approximately 8-wk intervals over 109 wk. Tibial nerve alpha-tocopherol concentrations were measured at 65 and 109 wk; adjacent sections were examined for histologic changes. In the two -E dogs, plasma alpha-tocopherols declined linearly on a semilog plot to < 0.1 microgram/mL by 109 wk. Plasma alpha-tocopherol concentrations were depleted to half of the initial concentrations in approximately 87 d. Adipose tissue alpha-tocopherol concentrations (based on wet weight, cholesterol or triglyceride) also declined linearly on semilog plots, and were depleted to half of the initial concentrations in approximately 120 d. Tibial nerve alpha-tocopherols (ng/microgram cholesterol) in -E dogs decreased to 16% of average +E at 65 wk, and to 2% at 109 wk. Neurologic examinations, histologies and nerve conduction velocities were normal in all dogs throughout the study. Our results demonstrate in dogs that depletion of plasma, adipose tissue and nerve alpha-tocopherol precedes histologic and functional changes in peripheral nerves during vitamin E deficiency.     

Effects of moderate-dose versus high-dose trimethoprim on serum creatinine and creatinine clearance and adverse reactions

The effects of a 10-day course of moderate-dose (10 mg/kg/day) or high-dose (20 mg/kg/day) trimethoprim therapy on serum creatinine, measured creatinine clearance, urinary creatinine excretion, and serum folate were studied in 20 healthy volunteers. Serum creatinine concentrations increased significantly during trimethoprim therapy, began to decrease near day 10, and returned to baseline during the washout phase at both dosage levels. At the same time, measured creatinine clearance and urine creatinine changed in the opposite direction. No clinical or statistical differences were noted between changes in the moderate- versus the high-dose phases. Serum folate concentration decreases during high-dose trimethoprim therapy were statistically significant. Adverse drug reactions in the two groups were statistically different during the first study period, with the high-dose group having a 75% incidence rate and the moderate-dose group having an 11% incidence rate (P < 0.02). Serum creatinine, measured creatinine clearance, and urinary creatinine excretion demonstrated statistically, but not clinically, significant changes during trimethoprim therapy. In addition, high-dose trimethoprim caused significantly more adverse drug reactions than moderate-dose trimethoprim in normal volunteers.     

Cardiopulmonary bypass-induced changes in plasma concentrations of propofol and in auditory evoked potentials

Unbound, rather than total, plasma concentrations may be related to the anaesthetic action of propofol. Therefore, we measured plasma concentrations of propofol and recorded Nb wave latencies of auditory evoked potentials (AEP) during continuous infusion of propofol in 15 patients undergoing coronary artery bypass grafting (CABG) surgery. After induction of anaesthesia with fentanyl, propofol was infused continuously at a rate of 10 mg kg-1 h-1 for 20 min, and then the rate was reduced to 3 mg kg-1 h-1. Administration of heparin before cardiopulmonary bypass (CPB) did not affect total or unbound propofol concentration. Initiation of CPB decreased mean total propofol concentration from 2.6 to 1.7 micrograms ml-1 (P < 0.01). Simultaneously, mean unbound propofol concentration remained at 0.06 micrograms ml-1 because of a slight increase in the mean free fraction of plasma propofol (from 2.3 to 3.5%; P > 0.05). During hypothermic CPB, mean total propofol concentration increased to concentrations measured before bypass (to 2.1 micrograms ml-1; P > 0.05 vs value before CPB) and the mean unbound propofol concentration was at its highest (0.07 microgram ml-1; P < 0.05 vs value before heparin). After CPB and administration of protamine, the mean total propofol concentration remained lowered (1.7 micrograms ml-1; P < 0.05 vs value before heparin) and the mean unbound propofol concentration returned to the level measured before heparin (P < 0.001 vs value during hypothermia). The latency of the Nb wave from recordings of AEP increased after induction of anaesthesia, reached its maximum during hypothermia and was prolonged during the subsequent phases of the study. The latency of the Nb wave did not correlate with total or unbound propofol concentration. We conclude that the changes in total and unbound concentrations of plasma propofol were not parallel in patients undergoing CABG. During CPB or at any other time during the CABG procedure, the unbound propofol concentration did not decrease and Nb wave latency was prolonged compared with baseline values measured after induction of anaesthesia before the start of CPB.     

Insulin-like growth factor I and insulin-like growth factor binding protein 3 as determinants of blood hemoglobin concentration in healthy subjects

We studied the serum concentrations of IGF-I, IGF-binding protein 3 (IGFBP-3), and testosterone in relation to blood Hb in 60 healthy prepubertal or early pubertal boys twice, with a 9-mo interval. Serum IGF-I and testosterone levels were measured by RIA, and serum IGFBP-3 was measured by monoclonal immunofluorometric assay. Positive correlations were observed between the concentrations of blood Hb and serum IGF-I at the first examination (r = 0.36, p = 0.008) and Hb and IGFBP-3 at both examinations (r = 0.53, p < 0.001, and r = 0.39, p = 0.003). No association between Hb and testosterone concentrations was found. Our results show that blood Hb is positively correlated to serum IGF-I and IGFBP-3 levels, indicating indirectly the involvement of growth hormone in the regulation of physiologic Hb concentration. Because no association was found between Hb and testosterone concentrations, this may indicate that the role of androgens in erythropoiesis may be different at different stages of puberty. It is concluded that the IGF system may be involved in the rise of Hb level during early puberty.     

Effect of different storage temperatures, sample collection procedures and immunoassay methods on osteocalcin measurement

The apparent instability of measured osteocalcin has been reported as method-dependent and related to preanalytical variables such as storage temperature, and the use of anticoagulants and protease inhibitors. The aim of this study was to determine a sample collection procedure which minimised osteocalcin degradation. Blood samples from five normal individuals were collected with or without anticoagulants and protease inhibitors (heparin, EDTA, or heparin and aprotinin) and stored at 4 degrees C, -20 degrees C or -70 degrees C for up to 7 days, 28 days and 90 days respectively. Osteocalcin was measured by both a monoclonal EIA specific for intact osteocalcin and a bovine polyclonal RIA. Osteocalcin concentrations in serum and EDTA-treated samples significantly decreased by 40% (P < 0.001) with the ELISA and 72% (P < 0.001) with the RIA after 7 days storage at 4 degrees C. Similar falls were documented in these samples when stored at -20 degrees C and -70 degrees C and measured by the ELISA. Minimal changes in osteocalcin immunoreactivity were observed in either assay when heparin-treated plasma with or without aprotinin was stored at -20 degrees C or -70 degrees C for up to 90 days. The apparent instability of measured osteocalcin can be minimised using these conditions.     

The level of the collagen cross-link pyridinoline reflects the improvement of cutaneous lesions in one case of skin alveolar echinococcosis

Cutaneous parasitic lesions, associated with a dense fibrous reaction, markedly improved under albendazole treatment in one case of supraumbilical skin localization of alveolar echinococcosis. Since collagen cross-linking increases during fibrogenesis and contributes to the stability of fibrotic lesions, we monitored the level of the cross-links pyridinoline and pentosidine in skin lesions from this patient to determine if they would reflect the changes occurring during treatment. We looked at the deposition of cross-linked type I collagen by immunohistochemistry and also measured the serum concentrations of pentosidine and of a fragment of type I collagen (ICTP), which contains a site of pyridinoline formation. Albendazole treatment did not affect either the collagen content of skin lesions or the serum concentrations of ICTP and pentosidine, but it led to a pronounced decrease in pyridinoline level concomitant with the disappearance, observed by immunohistochemistry, of extensively cross-linked fibrotic type I collagen. The follow-up of collagen cross-linking by pyridinoline in skin tissue thus appears to be useful in reflecting the improvement of fibrotic skin diseases during therapy.     

Effects of the naturally occurring food mycotoxin ochratoxin A on brain cells in culture

The potential of ochratoxin A (OTA) to damage brain cells was studied by using a three-dimensional cell culture system as model for the developing brain. Aggregating cell cultures of foetal rat telencephalon were tested either during an early developmental period, or during a phase of advanced maturation, over a wide range of OTA concentrations (0.4 nM to 50 microM). By monitoring changes in activities of cell type-specific enzymes (ChAt and GAD, for cholinergic and GABAergic neurones, respectively, GS for astrocytes and CNP for oligodendrocytes), the concentration-dependent toxicity and neurodevelopmental effects of OTA were determined. OTA proved to be highly toxic, since a 10-day treatment at 50 nM caused a general cytotoxicity in both mature and immature cultures. At 10 nM of OTA, cell type-specific effects were observed: in immature cultures, a loss in neuronal and oligodendroglial enzyme activities, and an increase in the activity of the astroglial marker glutamine synthetase were found, Furthermore, at 2 and 10 nM of OTA, a clustering of microglial cells was observed. In mature cultures, OTA was somewhat less potent, but caused a similar pattern of toxic effects. A 24 h-treatment with OTA resulted in a concentration-dependent decrease in protein synthesis, with IC50 values of 25 nM and 33 nM for immature and mature cultures respectively. Acute (24 h) treatment at high OTA concentrations (10 to 50 microM) caused a significant increase in reactive oxygen species formation, as measured by the intracellular oxidation of 2',7'-dichlorofluorescin. These results suggest that OTA has the potential to be a potent toxicant to brain cells, and that its effects at nanomolar concentrations are primarily due to the inhibition of protein synthesis, whereas ROS seem not to be involved in the toxicity mediated by a chronic exposure to OTA at such low concentrations.