Membranes from the neuroblastoma × embryonic retina cell hybrid cell line, N18-RE-105, bind l-[3H]glutamate with a pharmacologic profile consistent with a 'quisqualate-type' brain l-glutamate receptor. We describe here the cytotoxic effect of l-glutamate receptor agonists on intact N18-RE-105 cells. Cytotoxicity was quantitated by measurement of the release of the cytosolic enzyme, lactate dehydrogenase, into the culture medium after addition of l-glutamate and its analogs to the cell culture medium. l-Glutamate (10 mM) and its confirmationally restricted analogs, quisqualate (1 mM) and ibotenate (10 mM), caused cell lysis. In contrast, similar analogs which do not bind to N18-RE-105 cell membranes (kainic acid, N-methyl-d,l-aspartic acid and γ-aminobutyric acid) were not cytotoxic. l-Glutamate-induced cytotoxicity was eliminated when calcium-free medium was used. Addition of inorganic or organic calcium channel antagonists also reduced the cytotoxicity of l-glutamate, even when 1.8 mM calcium was present in the medium. Cadmium chloride (10 μM) completely blocked l-glutamate toxicity, whereas manganese chloride (150 μM) and lanthanum chloride (25 μM) reduced toxicity by greater than 50%. Dihydropyridine voltage-sensitive calcium channel agonists or antagonists, had little or no significant effect on l-glutamate-induced toxicity. In contrast, the verapamil derivatives, D600 and D888, and the diltiazem derivative, MDL 12,330A reduced l-glutamate toxicity by greater than 50%. These results suggest that a subtype of voltage-sensitive calcium channels is involved in the mechanism of l-glutamate receptor mediated cytotoxicity in this cell line.
Abstract: A growing literature indicates that blood levels of the hormone melatonin may have important implications for human health and wellbeing. Melatonin is synthesized and released into the general circulation at night, however, and it is seldom feasible to draw blood samples at night in epidemiological studies. There is some evidence that levels of urinary melatonin and of 6‐sulfatoxymelatonin (aMT6s), the major metabolite of melatonin, accurately reflect nocturnal plasma melatonin. If this is the case, urinary assays could be powerful tools for epidemiological studies. A laboratory‐based study was performed to examine the relationships between nocturnal plasma melatonin, morning urinary melatonin, and morning urinary aMT6s levels in 78 men. The relationship between total nocturnal plasma melatonin and both urinary aMT6s corrected for creatinine and urinary melatonin is significant. Combining the two urinary measures accounts for 72% of the variance in total plasma melatonin. Peak nocturnal plasma melatonin also was significantly related to urinary melatonin and to aMT6s. The urinary measures show good sensitivity and specificity in identifying individual differences in nocturnal plasma melatonin levels. These results support the inclusion of morning urine samples to assess the contribution of the hormone melatonin in occupational or residential studies involving healthy, young men.
Heart rate variability (HRV), a noninvasive indicator of autonomic control of cardiac activity, is predictive of long-term cardiac morbidity and mortality. Epidemiologic research suggests that occupational exposure to power-frequency magnetic fields may be associated with autonomically mediated cardiac mortality. Results from our laboratory studies of humans exposed to 60-Hz magnetic fields overnight, however, are inconsistent. HRV is altered in some studies but not others. To clarify this, the pooled data from seven studies involving 172 men were analyzed to test specific hypotheses concerning this inconsistency. After analysis, we excluded a) measurement drift or instability over time because HRV was stable under sham-exposed conditions across all studies; b) inadequate statistical power or failure to maintain double-blind controls; c) differences in field intensity (28.3 vs. 127.3 microT) or exposure pattern (intermittent versus continuous) as main effects; or d) the inclusion of individuals sensitive to magnetic field exposure in some studies but not others. Four separate analytic techniques failed to identify a valid subpopulation of sensitive individuals. In some studies, however, hourly blood samples were collected using an indwelling venous catheter. HRV alterations occurred during intermittent exposure in these studies (p < 0.05) but not in similar studies without blood sampling. This result suggests a field interaction with modest arousal or disturbance. Because HRV is tightly coupled to electroencephalographic activity during sleep, these results are physiologically plausible and suggest that HRV alterations during exposure to magnetic fields may occur when accompanied by increases in physiologic arousal, stress, or sleep disturbance.
Recent advances enable one to apply numerical techniques to anatomically-correct human models to compute current densities and electric fields in tissue due to exposure to electric fields, magnetic fields, or contact currents. These methods have proved to be informative in estimating exceedance of basic restrictions prescribed by exposure guideline organizations. To date, the analyses have been conducted with a resolution on the order of millimeters. However, these techniques have future roles to play at higher levels of resolution at those sites in target tissues suspected of transducing local electric fields into biological responses. Two specific cases in which high resolution “microdosimetry” would yield value involve (a) residential settings and childhood leukemia and (b) worker exposure and cardiovascular disease. Recent research suggests that residential contact currents on the order of microamperes can produce biologically significant dose (expressed as the local electric field) to the bone marrow of a child. Microdosimetry would focus on pluripotent progenitor cells resident in the marrow compartment, as well as anatomic features that distinguish a child’s from an adult’s marrow. Laboratory and epidemiologic research has suggested that magnetic field exposure may affect heart rate variability, a measure reflective of autonomic nervous system control of cardiac activity. Given the physical attributes of the central nervous system and the sites that could serve as substrates for field interactions, future microdosimetry addressing heart rate variability effects may be well-advised to focus on the electrically excitable dendritic arborizations of neurons. In both cases, microdosimetry will help shed light on primary interactions in tissue.
