Environmental PCB exposure and risk of endometriosis.

BACKGROUND: Hormonally active environmental agents have recently been associated with the development of endometriosis. METHODS: We undertook a study to assess the relationship between endometriosis, an estrogen-dependent gynaecological disease, and 62 individual polychlorinated biphenyl (PCBs) congeners. We enrolled 84 eligible women aged 18-40 years undergoing laparoscopy for study, which included an interview and blood specimen (n=79; 94%). Thirty-two women had visually confirmed endometriosis at laparoscopy while 52 did not. Blood specimens were run in batches of 14 including four quality control samples for toxicological analysis. Each PCB congener was adjusted for recovery; batch-specific reagent blanks were subtracted. All PCB concentrations were log transformed and expressed in ng/g serum first as a sum and then as tertiles by purported estrogenic or anti-estrogenic activity of PCB congeners. RESULTS: Using unconditional logistic regression analysis, a significantly elevated odds ratio (OR) was observed for women in the third tertile of anti-estrogenic PCBs [OR 3.77; 95% confidence interval (CI) 1.12-12.68]. Risk remained elevated after controlling for gravidity, current cigarette smoking and serum lipids (OR 3.30; 95% CI 0.87-12.46). CONCLUSIONS: These data suggest that anti-estrogenic PCBs may be associated with the development of endometriosis.

Effect of vanadium(IV) compounds in the treatment of diabetes: in vivo and in vitro studies with vanadyl sulfate and bis(maltolato)oxovandium(IV).

Vanadyl sulfate (VOSO(4)) was given orally to 16 subjects with type 2 diabetes mellitus for 6 weeks at a dose of 25, 50, or 100 mg vanadium (V) daily [Goldfine et al., Metabolism 49 (2000) 1-12]. Elemental V was determined by graphite furnace atomic absorption spectrometry (GFAAS). There was no correlation of V in serum with clinical response, determined by reduction of mean fasting blood glucose or increased insulin sensitivity during euglycemic clamp. To investigate the effect of administering a coordinated V, plasma glucose levels were determined in streptozotocin (STZ)-induced diabetic rats treated with the salt (VOSO(4)) or the coordinated V compound bis(maltolato)oxovandium(IV) (abbreviated as VO(malto)(2)) administered by intraperitoneal (i.p.) injection. There was no relationship of blood V concentration with plasma glucose levels in the animals treated with VOSO(4), similar to our human diabetic patients. However, with VO(malto)(2) treatment, animals with low plasma glucose tended to have high blood V. To determine if V binding to serum proteins could diminish biologically active serum V, binding of both VOSO(4) and VO(malto)(2) to human serum albumin (HSA), human apoTransferrin (apoHTf) and pig immunoglobulin (IgG) was studied with EPR spectroscopy. Both VOSO(4) and VO(malto)(2) bound to HSA and apoHTf forming different V-protein complexes, while neither V compound bound to the IgG. VOSO(4) and VO(malto)(2) showed differences when levels of plasma glucose and blood V in diabetic rodents were compared, and in the formation of V-protein complexes with abundant serum proteins. These data suggest that binding of V compounds to ligands in blood, such as proteins, may affect the available pool of V for biological effects.

Relation of Lake Ontario fish consumption, lifetime lactation, and parity to breast milk polychlorobiphenyl and pesticide concentrations.

Lactating female members and spouses of male members of the New York State Angler Cohort who agreed to provide breast milk samples were the subjects of this study. Questionnaires were provided to participants focusing on Lake Ontario fish consumption, reproductive history, and lactation history. Milk samples were analyzed for 77 polychlorinated biphenyls (PCB) congeners, 1,1-dichloro-2,2-bis (p-chlorophenyl)-ethylene (DDE), a metabolite of dichlorodiphenyltrichloroethane (DDT), hexachlorobenzene (HCB), and 1,1a,2,2,3,3a, 4,5,5,5a,5b,6-dodecachlorooctahydro-1,3, 4-methano-1H-cyclobuta[cd]pentalene (Mirex). The percentage of samples with quantifiable levels, above the limit of detection (LOD), varied among the individual congeners from 10 to 100%. Nine PCB congeners (designated by their IUPAC No.) and DDE were found in all of the 100 samples analyzed. These include the following, in decreasing order of concentration: DDE>153>138>180>118>187>188>177>200. Total PCB concentrations were estimated by taking the sum of the concentrations of all PCB congeners (up to 77 congeners) above their respective LOD in a given sample. PCB concentrations increased with increasing concentration of milk lipid. Lipid adjusted PCB concentrations increased as a function of maternal age. PCB congener profiles in milk favored the higher chlorinated congeners, with the four highest congeners having 5 to 7 chlorine atoms. Fish eaters had a significantly higher level of several major PCB congeners with congeners 153 and 138 being 1.36 and 1.34 times higher, respectively. PCB and DDE concentrations, expressed on a lipid basis, varied inversely with parity. The total number of months of lifetime lactation varied inversely with the total PCB concentration in breast milk. A similar relationship was evident for DDE. These data are of use for risk assessment in estimating the relative exposure to these environmental contaminants in breast fed infants whose mothers consumed contaminated Lake Ontario fish.

Comparison of PCB congeners and pesticide levels between serum and milk from lactating women.

Samples of blood and milk were obtained from lactating women participating in the New York State Angler study. A total of seven women gave one blood and one milk sample at time intervals between blood and milk collection different for each woman. The time between samples varied from 3 to 318 days. One subject provided a second milk sample 219 days after the first milk sample. The samples were analyzed for 69 PCB congeners, DDE (a metabolite of DDT), Mirex, and hexachlorobenzene (HCB). Lipid content was determined by gravimetric analysis. The congener profiles in serum and milk were similar for each individual but different among all subjects. The sum of the concentrations of the congeners present above the limit of detection was used to estimate the total PCB concentration that was in the range of 2.6 to 5.8 ng/g of serum and 3.5 to 14.1 ng/g of milk. The ratio of serum to milk concentrations varied from 0.18 to 1.66 with a mean of 0.65+/-0.49 showing no consistency among individuals prior to adjusting the data for lipid content. The total PCB levels normalized for lipid content were 320-728 ng/g of serum lipid and 239-428 ng/g of milk lipid. The range of the lipid adjusted serum/milk ratio was 1.1 to 2.8 and the mean+/-SD serum/milk ratio was 1.9+/-0.5. The ranges of lipid adjusted serum concentration of DDE, HCB, and Mirex were 95 to 591, 8 to 48, and 3 to 29 ng/g lipid, respectively. The ranges of lipid adjusted milk concentration of DDE, HCB, and Mirex were 90 to 577, 11 to 22, and 1 to 10 ng/g lipid, respectively. For DDE, HCB, and Mirex, the means of the individual lipid adjusted serum to milk ratios were 1.5+/-0.7, 2.5+/-1.5, and 5. 3+/-4.6, respectively. Considerable differences were found among lipid adjusted concentrations of these environmental pollutants in serum and milk samples from the same individual. This suggests that body burden estimates in lactating women using different matrices may not be equivalent even when lipid adjusted values are used.

Mercury as a potential hazard for the dental practitioner.

Mercury has been used for centuries for medical, chemical, metallurgical and electrical applications. It is an element of mystery, which in its metallic form is an enticing silvery liquid that can be as fascinating as it is dangerous. Its use in dental amalgam has a potential for continuous occupational exposure of dental practitioners to mercury vapor. It is imperative that the dental practitioner understands the hazards associated with the use of mercury, and controls exposures to prevent the development of any untoward effects. This article provides an overview of the toxicology of the different forms of mercury to which human exposure occurs and addresses safety issues associated with mercury vapor, the primary form of mercury encountered in the practice of dentistry.

The biochemical toxicology of 1,3-difluoro-2-propanol, the major ingredient of the pesticide gliftor: the potential of 4-methylpyrazole as an antidote.

Administration to rats of 1,3-difluoro-2-propanol (100 mg kg-1 body weight), the major ingredient of the pesticide gliftor, resulted in accumulation of citrate in the kidney after a 3 hour lag phase. 1,3-Difluoro-2-propanol was found to be metabolized to 1,3-difluoroacetone and ultimately to the aconitate hydratase inhibitor (-) erythrofluorocitrate and free fluoride. The conversion of 1,3-difluoro-2-propanol to 1,3-difluoroacetone was found to be catalyzed by an NAD(+)-dependent alcohol dehydrogenase, while the defluorination was attributed to microsomal monooxygenase activity induced by phenobarbitone and inhibited by piperonyl butoxide. 4-Methylpyrazole was found to inhibit both of these processes in vitro and when administered (90 mg kg-1 body weight) to rats, 2 hours prior to 1,3-difluoro-2-propanol, eliminated signs of poisoning, prevented (-) erythrofluorocitrate synthesis, and markedly decreased citrate and fluoride accumulation in vivo. 4-Methylpyrazole also appeared to diminish (-) erythrofluorocitrate synthesis from fluoroacetate in vivo, and this was attributed to its capacity to inhibit malate dehydrogenase activity. The antidotal potential of 4-methylpyrazole and the potential for 1,3-difluoro-2-propanol to replace fluoroacetate (compound 1080) as a vertebrate pesticide is discussed.

Standardization of a method for the routine analysis of polychlorinated biphenyl congeners and selected pesticides in human serum and milk.

A methodology is presented for the routine determination of specific polychlorinated biphenyl (PCB) congeners in serum and milk samples. The procedures include standardized extraction, cleanup and quantitation by high-resolution gas chromatography (GC) and comprehensive quality assurance program to minimize systematic and erratic errors. The analyses of 68 PCB congeners and three pesticides, p,p1-dichloro diphenyl dichloro ethylene (DDE), hexachlorobenzene (HCB), and Mirex, at part-per-billion levels include the addition of surrogate congener standards (IUPAC isomers #46 and #142), extraction with hexane after protein precipitation, cleanup with Florisil, and analysis by GC with capillary column and electron capture detection. Quantitation is based on calibration standards and response factors using isomers #30 and #204 as internal standards. The quality control activities consist of analyses of samples in batches of 6 to 10 simultaneously with quality control (QC) samples. The quality assurance program checks that the procedures are under control by the use of control charts and set the criteria for data acceptability. The detection limits for the congeners and pesticides associated with the analyses of 500 serum samples and of 100 milk samples are reported. In addition, typical profiles of congener distribution in both matrices are illustrated.

Altered intrarenal accumulation of mercury in uninephrectomized rats treated with methylmercury chloride.

We tested the hypothesis that the intrarenal accumulation of mercury in rats treated with methylmercury is altered significantly as a result of unilateral nephrectomy and compensatory renal growth. Renal accumulation of mercury was evaluated by radioisotopic techniques in both uninephrectomized (NPX) and sham-operated (SO) rats 1, 2, and 7 days after the animals received a nonnephrotoxic intravenous dose of methylmercury chloride (5 mg/kg Hg). At all times studied after the injection of the dose of methylmercury, the renal accumulation of mercury (on a per gram kidney basis) was significantly greater in the NPX rats than that in the SO rats. The increased accumulation was due to a specific increase in the accumulation of mercury in the outer stripe of the outer medulla. Renal cortical accumulation of mercury was similar in both the NPX and SO rats. The percentage of the administered dose of mercury that was present in the total renal mass of the NPX and SO rats ranged between 5 and 15, depending on the day that the renal accumulation was studied. Approximately 40-50% of the total renal burden of mercury in both the NPX and SO rats was in the inorganic form. However, only less than 1% of the mercury in blood was in the inorganic form at the three times accumulation was studied. Very little mercury was excreted in the urine by either the NPX or SO rats. Only about 2 to 3% of the administered dose of mercury was excreted in the urine in 7 days. By contrast, the cumulative fecal excretion of mercury over 7 days was substantial in the NPX and SO rats, and significantly more mercury was excreted in the feces by the NPX rats (about 19% of the dose) than by that in the SO rats (about 16% of the dose). In conclusion, our findings indicate that unilateral nephrectomy and compensatory renal growth cause a significant increase in the accumulation of mercury in the renal outer stripe of the outer medulla in rats exposed to methylmercury. In addition, the findings indicate that the fecal excretion of mercury is also significantly increased.

Primary cultures of rabbit renal proximal tubule cells. III. Comparative cytotoxicity of inorganic and organic mercury.

The present study further developed primary cultures of rabbit renal proximal tubule cells (RPTC) as an in vitro model to study chemical-induced toxicity by investigating the comparative cytotoxicity of mercuric chloride (HgCl2) and methyl mercury chloride (CH3HgCl) to RPTC. Confluent monolayer cultures of RPTC exposed to HgCl2 and CH3HgCl for 24 hr exhibited a concentration-dependent loss in cell viability at culture medium concentrations greater than 25 and 2.5 microM, respectively. Vital dye exclusion was a more sensitive indicator of cytotoxicity than the amount of lactate dehydrogenase activity, alkaline phosphatase activity, N-acetylglucosaminidase activity, and protein content remaining on the culture dish. On the basis of vital dye exclusion, HgCl2 was less toxic to proximal tubule cells in culture than CH3HgCl after 24 hr of exposure, whether cytotoxicity was based on LC50 values (34.2 microM HgCl2 vs 6.1 microM CH3HgCl) or total cellular mercury uptake (4.6 nmol Hg2+/10(5) cells vs 1.25 nmol CH3Hg+/10(5) cells). Differences in the extent and rate of metal uptake were also evident. Maximum cellular uptake of Hg2+ occurred within 6-24 hr after exposure and was not concentration-dependent, whereas maximum uptake of CH3Hg+ occurred within 3 hr of exposure and was concentration-dependent. The intracellular distribution of both mercurials between acid-soluble and acid-insoluble binding sites also differed. At noncytotoxic concentrations of HgCl2 (0.04-5 microM), intracellular Hg2+ bound increasingly to acid-soluble binding sites as a function of time, from 15-30% after 6 hr of exposure to 40-60% after 72 hr of exposure. However, at subcytotoxic (25 microM) and cytotoxic (34.2 microM) concentrations, Hg2+ binding to acid-soluble binding sites remained constant at approximately 30-40% for 6, 12, 24, and 72 hr after exposure. In contrast, only 20% of total cellular CH3Hg+ was bound to acid-soluble binding sites after exposure to 0.039 to 6.1 microM CH3HgCl for 6, 12, and 24 hr. Total cellular glutathione content was unaffected after exposure to 0.04-5 microM HgCl2 and 0.039-6.1 microM CH3HgCl, but was depleted 6 hr after exposure to 25 and 34.2 microM HgCl2. These results indicate that CH3HgCl was a more potent cytotoxicant to RPTC in primary culture than HgCl2. Furthermore, compared to Hg2+, the low binding of CH3Hg+ to acid-soluble binding sites and the absence of a redistribution of CH3Hg+ from acid-insoluble to acid-soluble binding sites appeared to contribute to its more potent toxicity to cultured cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Placental opioid-enhancing factor (POEF): generalizability of effects.

A substance in amniotic fluid and placenta (POEF for Placental Opioid-Enhancing Factor) has been shown to enhance opiate- or opioid-mediated analgesia in rats. Recent studies have only touched on the generalizability of the phenomenon. The present studies further tested the generalizability of the POEF effect: they examined sex specificity of the mechanism; whether POEF activity exists in afterbirth material of species other than the rat; whether POEF activity exists in tissue other than afterbirth material; whether POEF activity could be demonstrated after injection rather than ingestion of afterbirth material; and whether POEF enhances all opioid-mediated phenomena. We found that (a) POEF is effective in male rats as well as in female rats; (b) POEF activity exists in human and dolphin afterbirth material; (c) ingestion of pregnant-rat liver does not produce enhancement of opioid-mediated analgesia; (d) POEF does not seem to be effective when amniotic fluid is injected either IP or SC; and (e) POEF does not modify morphine-induced hyperthermia.

Acute toxicity and mutagenicity of the copper complex of pyruvaldehyde-bis (N-4-methylthiosemicarbazone), Cu-PTSM.

Cu-PTSM is a potential imaging agent for the heart and brain when labeled with either 64Cu or 62Cu. Unlabeled Cu-PTSM was evaluated for its acute toxicity and mutagenicity. Cu-PTSM had an i.v. LD50 of 26 mg kg-1 in the rat and 2 mg kg-1 in the rabbit. At necropsy, rats exhibited severely hemorrhagic lungs, histological findings of acute pulmonary congestion, hemorrhage and edema, and mild congestion in kidney, liver and brain. The rabbit displayed marked polymorphonuclear infiltration in alveoli, peribronchial and periarterial areas with marked macrophage hyperplasia, congestion and mild hemorrhage into alveolar spaces. No effects were found in kidney, liver, testes or brain. Administration of 2.16 micrograms kg-1 day-1 for 5 days per week for 2 weeks resulted in no changes in histopathology, hematology or clinical chemistry parameters. This daily dose is at least 300 times the diagnostic dose intended for use in man. Cu-PTSM was not mutagenic when tested in the absence of S9 supernatant, but elicited a weakly mutagenic response in the presence of S9. Since acute effects in the lung occur at doses approaching 300,000 times the diagnostic dose, it is highly unlikely that the clinical use of Cu-PTSM would result in any acute adverse effects.

Extracorporeal regional complexing haemodialysis treatment of acute inorganic mercury intoxication.

A 70-year-old white female presented approximately 24 h after ingesting three 475 mg tablets (1.425 g) of mercuric chloride in a suicide attempt. Acute renal failure necessitated the initiation of haemodialysis approximately 4 d after the ingestion. Treatment with BAL (2,3-dimercaptopropanol) resulted in only small increases in mercury output into dialysate. A new procedure involving the extracorporeal infusion of the chelating agent dimercaptosuccinic acid (DMSA) into the arterial blood line during haemodialysis was initiated. This procedure of Extracorporeal Regional Complexing Haemodialysis (ERCH) had been effective in increasing methylmercury removal in patients poisoned by contaminated grain. The first DMSA-ERCH procedure was performed 6 d after poisoning. There was a dramatic increase in mercury output into the dialysate. During three treatment sessions of 80 min each, 1189 micrograms of mercury were removed from the patient. The dialysed mercury represented the only mercury output since the patient was anuric and not producing faeces. DMSA-ERCH appears to be much more effective than BAL and haemodialysis in the treatment of acute inorganic mercury poisoning. The long interval between poisoning and initiation of treatment probably contributed to the patients ultimate demise, 28 d after poisoning. Efficacy of the DMSA-ERCH procedure for inorganic mercury poisoning is likely to be improved as the interval between exposure and treatment is reduced.

Primary cultures of rabbit renal proximal tubule cells: II. Selected phase I and phase II metabolic capacities.

Specific characteristics of cells vary as a function of time in culture. We have determined the stability of selected Phase I and Phase II biotransformation capacities in rabbit renal proximal tubule cells in primary culture. When grown in hormonally-defined medium, proximal tubule cells lost Phase I metabolic capacity. Cytochrome P-450 content and associated mixed-function oxidase activities present in kidney cortex microsomes were not detectable after 14 days in culture. Phase II glutathione-dependent metabolic functions were well retained in cultured cells compared with freshly isolated proximal tubules (FIPT). Cellular total glutathione content was 2.8 mug/mg protein in FIPT compared with approximately 10 mug/mg protein in stable confluent cultures. A higher total glutathione content of 20.6 mug/mg was noted in preconfluent cultures. The glutathione redox state was initially perturbed in FIPT with 37% of the total glutathione present found in its oxidized form. Tubule cells recovered to a normal ratio (6-13% of total glutathione in the oxidized form) while in culture. The glutathione S-transferase activity in 4-day-old cells in culture was reduced to 50% of the 4 U/mg protein level found in FIPT. No appreciable further decline in glutathione S-transferase activity was detected during 15 days in culture. The level of gamma-glutamyl-transpeptidase (a brush-border enzyme necessary for glutathione uptake into proximal tubule cells) declined from 1499 mU/mg protein in homogenates of FIPT to 636 mU/mg in homogenates of 8-day-old cultured cells. A further decline in activity occurred during the next 7 days in culture. In conclusion, although Phase I metabolic functions were diminished in primary cultured rabbit proximal tubule cells, Phase II metabolic functions were retained at levels comparable with FIPT and well above those found in several established kidney cell lines.

Primary cultures of rabbit renal proximal tubule cells: I. Growth and biochemical characteristics.

Before the usefulness of a new in vitro model can be ascertained, the model must be properly defined and characterized. This study presents the growth rate and biochemical characteristics of rabbit renal proximal tubule cells in primary culture over a 2-wk culture period. When grown in a hormonally defined, antibiotic-free medium these cells form confluent monolayer cultures within 7 d after plating. Multicellular dome formation, an indicator of transepithelial solute transport, was expressed after confluent cultures were formed. The activity of the cytosolic enzyme, lactate dehydrogenase, and the lysosomal enzyme, N-acetyl-glucosaminidase, increased 14- and 2-fold during the first 8 d of culture, respectively. In contrast, the activity of a brush border enzyme, alkaline phosphatase, decreased 85% within the first 8 d of culture. Release of these enzyme markers into the culture medium, which are routinely used to measure cytotoxicity, stabilized after 8 d in culture. The ratio of cellular protein to DNA changed according to the state of cellular growth. Values rose from 0.035 mg protein/micrograms DNA in preconfluent cultures to 0.059 mg protein/micrograms DNA in confluent cultures. These results document the characteristics of a primary proximal tubule cell culture system for future studies in in vitro toxicology.

Cell specific enzyme markers as indicators of neurotoxicity: effects of acute exposure to methylmercury.

In order to assess the sensitivity of several cell specific enzyme markers (tyrosine hydroxylase (TH), glutamic acid decarboxylase, choline acetyltransferase, glutamine synthetase (GS), neuron specific and non-neuronal enolase and 2',3'-cyclic nucleotide phosphohydrolase (CNP] as indices of neurotoxicity, changes in their activities were monitored after rats were treated with two doses of the neurotoxic agent, methylmercury chloride (MMC). Comparisons were also made of any histopathological changes occurring in the tissues examined. At the low dose rate (3.36 mg Hg/kg, po, for 14 days), the rats exhibited less body weight gain compared to untreated animals. No change in either the neuronal or noneuronal enzyme markers was observed in brain but a significant increase in the myelin marker, CNP, and total enolase activity was seen in the optic nerve. Morphological evaluation by light microscopy indicated no discernible neuronal lesions in MMC-exposed animals. At the higher MMC dose (7.05 mg Hg/kg, po, for 7 days), there was about a 20% loss in the body weight of treated animals and partial hind limb paralysis was observed. Of all the neuronal marker enzymes examined, only TH was found to be decreased in the striatum. The proliferating astroglial marker, GS, was elevated only in the cerebellum. CNP was found to be decreased in both the optic and sciatic nerve. As in the lower dose group no pathological changes were observed at the light microscopic level in the brain of MMC-treated rats. These data suggest that of the cell specific marker enzymes studied, GS in the cerebellum and TH in the striatum may be useful biochemical markers for the neurotoxic action of MMC.

Effect of chronic exposure to hexachlorophene on rat brain cell specific marker enzymes.

The neurotoxicity associated with chronic exposure to hexachlorophene (HCP) was evaluated by measuring the activity of seven cell specific marker enzymes in brain and by comparing these measurements to morphological changes analyzed by light microscopy. Animals were divided into two groups, the experimental group received HCP at a daily dose of 20 mg/kg p.o. for 53 consecutive days whereas the control group received an equivalent amount of the vehicle only. HCP produced no change in the rate of gain in body weight nor did it produce a statistically significant change in brain weight. Furthermore, no overt abnormal neurological symptoms were observed at this level of exposure to HCP. The white matter throughout the brain was extensively vacuolated in the HCP-treated rats, imparting a spongiform structure which was absent in the white matter of the control animal brains. The data obtained reveal that chronic HCP treatment produce little change in any of the neuronal marker enzymes with the exception of a significant decrease in tyrosine hydroxylase activity in the striatum. Of the nonneuronal enzymes assayed, a significant increase in non-neuronal enolase, glutamine synthetase, and 2',3'-cyclic nucleotide phosphohydrolase was observed in the sciatic nerve, hippocampus and optic nerve, respectively.

Effect of short-term exposure to hexachlorophene on rat brain cell specific marker enzymes.

Seven cell specific marker enzymes in brain and optic nerve and morphological evaluation by light microscopy were used to characterize the neurotoxicity associated with exposure of rats to hexachlorophene (HCP; 40 mg/kg/day, po, for 9 days). In vitro exposure to HCP at concentrations up to 100 microM had no direct inhibitory effect on the marker enzymes, validating their use in evaluating brain function in vivo. Rats exhibited a reduction in body weight gain, weakness, and ataxia of the hind limbs by the ninth day of HCP exposure. At 24 hr following the last day of exposure to HCP, the activities of the three neuron specific enzymes, glutamic acid decarboxylase, tyrosine hydroxylase, and choline acetyltransferase, in rat brain were unchanged from those of the vehicle-treated control group. Of the two astroglial enzyme markers measured, a small but significant increase was observed in the activity of nonneuronal enolase in the cerebellum and glutamine synthetase in the hippocampus of HCP-treated rats. The optic nerve appeared to be the most sensitive tissue in that the activity of both the astroglial marker, nonneuronal enolase, and the myelin marker, 2',3'-cyclic nucleotide phosphohydrolase, was significantly decreased following HCP exposure. This decrease in enzyme activity is consistent with the histological observations demonstrating extensive vacuolization and edema in the optic nerve after exposure to HCP.

The toxicity and mutagenicity of S-3-iodo-N-(1-ethyl-2-pyrrolidinyl)methyl-2-hydroxy-6-methoxybenzamide (IBZM), a new CNS D-2 dopamine receptor imaging agent.

IBZM is one of several benzamide derivatives showing a high affinity for the CNS D-2 dopamine receptor. Carrier-free [123I]IBZM is potentially useful as a nuclear medicine imaging agent for investigating CNS D-2 dopamine receptor in humans. This study describes the acute toxicity of IBZM in the rat and rabbit, its subchronic toxicity in the rabbit and its mutagenicity measured by the Ames test. IBZM had a 24 hour LD50 of 400 mg/kg in the rat and 50 mg/kg in the rabbit when administered i.v. Deaths occurred within minutes of dosing. Some necrosis was evident at the injection site in IBZM treated animals which was not found in the controls. No gross or histological differences between experimental animals and controls were evident in surviving animals when necropsied 14 days after dosing. Repeated exposure of rabbits to IBZM at a total cumulative dose of 100,000 times the expected clinical dose revealed no consistent changes in hematology, blood chemistry, blood enzymes or tissue pathology. IBZM was not mutagenic in the modified Ames assay with or without metabolic activation in the TA98 and TA100 tester strains. It is therefore unlikely that acute adverse effects will be associated with the diagnostic use of this drug in man.

Effect of a course in cancer prevention on students' attitudes and clinical behavior.

The long-term effects of a first-year medical school course in disease prevention focusing on cancer were studied. The authors hypothesized that students' increased knowledge of disease prevention principles would result in more positive attitudes toward aggressive treatment and prevention of cancer and a more frequent use of disease prevention techniques in clinical settings. On two occasions one to three years after the course, groups of students who took the course and groups that did not take the course responded to a test of knowledge of cancer, a cancer attitude survey, a clinical practice survey, and an evaluation of the course's relevance. Also, their clinical behavior was observed and coded during videotaped interviews of patients. While the students who took the course were more knowledgeable about cancer than their peers who did not, they did not have significantly more positive attitudes; nor were they significantly more likely to use prevention techniques in clinical settings. However, the more knowledgeable students--both those who took the course and the control students--had more positive attitudes than the less knowledgeable students. Those students who considered the course relevant to their training were more knowledgeable and had more positive attitudes toward cancer prevention than those who did not consider it relevant.

Effect of L-(alpha S,5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid on urinary excretion of methylmercury in the mouse.

The gamma-GTP inhibitor L-(alpha S,5S)-alpha-amino-3-chloror-4,5-dihydro-5-isoxazoleacetic acid (AT-125) was administered to CBA/J mice pretreated with 203Hg-methylmercury (MM) (0.5 mg/kg) to determine whether increasing urinary GS in this strain would result in a simultaneously increased urinary elimination of MM. Doses at AT-125 ranging from 3.0 to 30 mg/kg increased urinary GS in a dose-related fashion. The peak effect was attained at 2 hr after injection. AT-125 (7.5 mg/kg) increased urinary GS to peak value of 450 microM. When this dose was administered to mice 24 hr after MM, neither the rate of decline in body burden of 203Hg nor the rate of 203Hg output into urine or feces varied significantly from control values. A dose of 15 mg of AT-125 per kg increased urinary GS to 1.0 mM and caused a significant increase in urinary excretion of 203Hg when compared to the 7.5-mg dose group. The greatest effect was seen at the 30-mg AT-125/kg dose which produced a 1.9 mM concentration of total GS in urine and a 2-fold increase in the urinary 203Hg excretion rate. The dose-dependent changes in urinary excretion elicited by AT-125 were paralleled by increased rates of decline of 203Hg body burden and decreased rates of excretion in the feces. The results suggest that urinary GS must approach the millimolar range before affecting a redistribution of MM across the luminal membrane.