Health effects of ingesting arsenic-contaminated groundwater.

Approximately 4% of private drinking water wells in central Outagamie and Winnebago counties contain naturally occurring arsenic at concentrations exceeding the present Environmental Protection Agency (EPA) standard of 50 parts per billion (ppb). This cross-sectional study assessed health status via a self-administered survey. Questions concerned water consumption and resulting neurological, gastrointestinal, and dermatological symptoms. Participants were recruited through a public information campaign recommending water testing through local health departments; the survey was distributed at the time the water sample was submitted. Participants were unaware of their arsenic water concentration before completing the survey. Surveys from 637 families--64% of all surveys distributed--representing 1,836 individuals were completed. Of those respondents, 452 (25%) consumed between 5 and 50 micrograms of arsenic per day through water consumption; 47 (3%) consumed more than 50 micrograms of arsenic per day. This sample size had 80% power to detect a two-fold difference in the prevalence of the health effects studied. At this power level, no significant associations between arsenic intake, estimated through drinking water consumption and water arsenic concentrations, and the health effects studied were established.

Mutagenic activity of nitro-substituted cyclopenta-fused polycyclic aromatic hydrocarbons towards Salmonella typhimurium.

Cyclopenta-fused isomers of pyrene and benz[a]anthracene, nitrated on the etheno bridge, were synthesized and tested in the Ames plate-incorporation assay. Since enzymatic reduction, if it occurs in these compounds, would form arylhydroxylamines which in turn would form highly stabilized arylnitrenium ions, we hoped to test the hypothesis that the direct-acting mutagenic activity of nitroPAH is correlated with the degree of stabilization of the electrophilic intermediate. We found that these compounds are mutagenic (1-9 rev/nmole in Salmonella typhimurium TA98) and do not require S9 activation. However, this activity is substantially lower than that of other nitroPAH of comparable size such as 1-nitropyrene (250-300 rev/nmole). The reasons for this comparative lack of activity are discussed with reference to current theories regarding structure-activity relationships of nitroPAH.

Cellular contribution to pH-mediated calcium flux in neonatal mouse calvariae.

Net calcium flux from cultured neonatal mouse calvariae into the culture medium is pH dependent, and acidified culture medium causes egress of calcium from bone. To determine whether calcium flux is mediated by pH effects on bone cell function, we cultured calvariae for 24 h with sodium azide, acetazolamide, parathyroid hormone (PTH), 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], or after three successive freeze-thaw cycles, treatments that would be expected to alter bone cell function. We recultured bones for 3 h with the respective treatment and measured calcium flux. Sodium azide and freeze-thaw cycles produced a net influx of calcium (JCa = -22 +/- 7 and -23 +/- 6 nmol X bone-1 X 3 h-1, respectively) compared with net efflux of control bones (JCa = 35 +/- 6) at a similar initial medium pH. Acetazolamide reduced net flux to 0 (JCa = 7 +/- 6). PTH and 1,25(OH)2D3 increased net calcium efflux from bone (JCa = 78 +/- 7 and 74 +/- 10, respectively). Despite changing net flux, the slope dependence of net flux on medium pH was the same in the control group and all five treated groups of bones. The similarity of slopes indicates that the pH dependence of net flux is not a result of pH acting on bone cells but probably an effect of altered mineral equilibria. The difference in net flux at similar pH indicates that calcium efflux is partially inhibited by acetazolamide and stimulated by both PTH and 1,25(OH)2D3.

Effect of pentobarbital on Na and Ca action potentials in an invertebrate neuron.

It has been suggested that Ca currents may be more sensitive to barbiturate blockade than Na currents. This hypothesis has been tested by comparing the effect of pentobarbital (PB) on the maximum rate of rise (V max) of Ca-depended and Na-dependent action potentials in cell R2 of the Aplysia abdominal ganglion. In Ca-free medium ([Na]o = 494 mM), V max of Na-spikes ranged from 50 to 100 V/s, while in Na-free medium ([Ca]o = 30 mM), V max of Ca-spikes ranged from 7 to 20 V/s. Under these conditions, Ca-spikes were 3-4 times more sensitive to barbiturate blockade than were Na-spikes. However, it was found that the sensitivity of Na- and Ca-spikes to PB dependent on V max of the spike prior to drug addition. V max was manipulated by altering the driving force on the current carrying cation; this was accomplished by changing the concentration of the cation in the bathing medium. Thus, Ca-spikes, elicited in media containing 10 mM Ca, were more sensitive to PB than were Ca-spikes elicited in 30 mM Ca. Likewise, the sensitivity of Na-spikes to PB could be altered by changing the external Na concentration, and consequently, V max. When the external Na and Ca concentrations were adjusted so that V max of Na- and Ca-spikes were similar, prior to drug addition, the PB dose-response curves for Na- and Ca-spikes overlapped. The mechanism accounting for the dependence of PB sensitivity on V max prior to drug addition remains unclear. However, the observation that PB dose-response curves for Na- and Ca-spikes are similar when V max of the spikes is similar, suggests that Na and Ca currents may be equally sensitive to PB in this particular cell.

Do identical activity patterns in fast and slow motor axons exert the same influence on the twitch time of cat skeletal muscle?

1. The fast-twitch flexor digitorum (the lateral head; equivalent to the flexor hallucis longus) and slow-twitch soleus muscles of the cat were denervated, and the two nerves immediately reunited to one or the other muscle. Contraction times of the dually reinnervated muscle were examined 9 weeks post-operatively in response to separate stimulation of its own and the foreign nerve. Over a 5 week period before the terminal experiment, a variety of artificial activity patterns was imposed on the two nerves.2. Following the dual-union operation, the flexor digitorum muscle was preferentially reinnervated by its own nerve. In contrast, the soleus muscle showed no evidence of preferential reinnervation.3. When neural activity was not artificially modified, the dually reinnervated flexor digitorum or soleus muscle showed faster contractions in response to stimulation of the flexor digitorum nerve than to stimulation of the soleus nerve.4. Following a 5 week period in which neural activity was virtually eliminated by cord transection or in which the two nerves were stimulated at the same frequency, the contraction times of the dually reinnervated soleus muscle were the same in response to stimulation of either nerve.5. In contrast, under the experimental conditions described above (cord transection or nerve stimulation), the dually reinnervated flexor digitorum muscle showed a significantly faster contraction in response to stimulation of its own nerve than to stimulation of the soleus nerve.6. It is concluded that, when neural activity is absent or identical in pattern, motoneurones normally innervating the fast- or slow-twitch muscles exert the same influence on contraction times of the soleus muscle.7. The dependence of contraction times of the dually reinnervated flexor digitorum muscle upon the type of the innervating motoneurone may be explained either by selective reinnervation of a particular group of muscle fibres or by different trophic substances emanating from the motoneurones.

Reaction of synapses on motoneurones to section and restoration of peripheral sensory connexions in the cat.

1. Monosynaptic excitatory post-synaptic potentials (e.p.s.p.s.) were recorded from triceps surae motoneurones of the cat 2-8 months after section of the medial gastrocnemius nerve whose regeneration into the muscle was prevented. In other animals, the cut nerve was reunited to the muscle with a post-denervation delay varying from 2 to 6 months, and the e.p.s.p.s were recorded 2 months later. The e.p.s.p.s were evoked by stimulation of the medial gastrocnemius nerve central to the site of the section or reunion. 2. The mean amplitudes of the e.p.s.p.s decreased with time after nerve section, virtually disappearing by the eighth post-operative month. The decrement of the e.p.s.p. amplitude occurred more quickly in soleus motoneurones than in medial or lateral gastrocnemius motoneurones. 3. The e.p.s.p.s evoked in medial or lateral gastrocnemius motoneurones from the cut medial gastrocnemius nerve returned to normal levels 2 months after reunion of the cut nerve even following a post-denervation delay of 6 months. However, in soleus motoneurones the e.p.s.p. recovery was incomplete. 4. The degree of functional motor reinnervation of the medial gastrocnemius muscle depended upon the post-denervation delay preceding the reunion operation. With a delay of 6 months before the reunion, the muscle showed no or only weak contractions in response to nerve stimulation. 5. Axotomized medial gastrocnemius motoneurones showed a significant decrease in conduction velocity and significant increase in the amplitude of overshoot of action potentials. The changes persisted even when the cut nerve was reunited to the muscle 2-6 months after denervation. 6. When the cut medial gastrocnemius nerve was reunited to the muscle 4 months after denervation, most of the sensory fibres, tested 2 months later, failed to respond to muscle stretch. 7. It is concluded that recovery of monosynaptic e.p.s.p.s following reconnexion of the cut nerve with its muscle does not require recovery of muscle activity, full restoration of sensory activity or the recovery of normal motoneurone properties. 8. It is also suggested that the degree to which central synaptic efficacy declines and recovers following section and regeneration of a peripheral nerve depends partly upon the type of motoneurone (fast phasic or slow tonic) with which the sensory fibres form synaptic connexions.

Membrane potentials in pinched-off presynaptic nerve ternimals monitored with a fluorescent probe: evidence that synaptosomes have potassium diffusion potentials.

1. Some physiological properties of tissue fractions from rat brain homogenates have been examined. Of the three fractions studied (presynaptic nerve terminals, mitochondria and fragmented membranes), only the nerve terminals (synaptosomes) have the ability to accumulate 42K from physiological salt solutions. 2. The ability to accumulate and retain K is lost if synaptosomes are exposed to very hypotonic solutions. The K uptake and total K content is reduced by ouabain and by inhibitors of glycolysis and oxidative phosphorylation. 3. These results suggest that synaptosomes in physiological saline accumulate K against a concentration gradient, and may have K diffusion potentials across their surface membranes. The voltage-sensitive fluorescent probe, 3,3'-dipentyl 2,2'-oxacarbocyanine (CC5), was used to test this possibility. 4. In the squid axon, the fluorescent emission of CC5 is directly proportional to membrane potential; depolarization causes an increase in fluorescence. 5. The fluorescence of synaptosomes ('synaptosome fluorescence') treated with CC5 is increased when [K]o is increased or [K]o is reduced; replacement of external Na by Li or choline has little effect on the synaptosome fluorescence. In quantitative terms, synaptosome fluorescence is proportional to log ([K]o plus 0-05[Na]o). Rb is about as effective as K in enhancing synaptosome fluorescence; Cs is about 1/4 as effective. The effect of increased [K]o is reversible. 6. The fluorescence data provide corroborative evidence that there is normally a large K gradient ([K]o smaller than [I]i) across the synaptosome surface membrane. The data suggest the [K]i may be in excess of 100 mM. 7. Replacement of Cl- by methylsulphate did not significantly affect the relationship between synaptosome fluorescence and [K]o, nor did removal of external Ca. 8. The fluorescence of CC5-treated mitochondria, membrane fragmnets, or lysed synaptosomes is unaffected by changes in the K concentration of the medium. 9. Veratridine and gramicidin D, both of which enhance Na permeability (PNa) in some intact tissues, increase synaptosome fluorescence when added to the standard medium. The increment is greatly reduced or abolished when external Na is replaced by choline. 10. If synaptosomes are first Na-loaded (by pre-treatment with cyanide + iodoacetate), and then placed in a choline medium, addition of gramicidin D significantly decreases fluorescence. This effect could be explained if, with [Na]o smaller than [Na]i, the increase in PNa causes the synaptosomes to hyperpolarize. 11. The veratridine-induced increase in synaptosome fluorescence was prevented by 3 times 10- minus 7M tetrodotoxin, which also blocks the depolarizing effect of veratridine in intact neurones. 12. The main conclusion is that synaptosomes may retain resting membrane potentials and the ability to increase Na permeability.