Biomedical subjects

D R Moore

Publications and source records attributed to D R Moore.

At least 55 records · Page 3Linked to original sources

PubMed1996

Topographic organization of projection from the parabigeminal nucleus to the superior colliculus in the ferret revealed with fluorescent latex microspheres.

Unilateral, discrete injections of red and green fluorescent latex microspheres or injections of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) were made into the ferret's superior colliculus (SC) to characterize the topographic organization of the projection from the parabigeminal nucleus (PBN). Retrograde labelling in the PBN revealed that this nucleus projects bilaterally to the SC, although the heaviest projection arises from the ipsilateral PBN. The PBN-SC projection demonstrates a highly ordered organization along the rostral-caudal axis; rostral PBN projects to rostral SC and caudal PBN projects to caudal SC. The caudoventral and rostrodorsal areas of the PBN project mainly to the ipsilateral and contralateral SC, respectively. The ipsilateral pathway terminates principally in the caudal region of the SC, while the contralateral projection terminates predominantly in rostral SC. Ipsilaterally, there are slightly more neurons, located mainly in the ventral PBN, that project to the lateral SC than those, located largely in the dorsal part of the nucleus, that target the medial SC. The contralateral PBN mainly projects to the rostrolateral quadrant of the SC. These results indicate that each quadrant of the SC is innervated principally by a restricted part of the PBN: the caudolateral quadrant, which receives the heaviest ipsilateral input, and the caudomedial quadrant are targeted predominantly by the ventral and dorsal portions, respectively, of the ipsilateral PBN; the rostrolateral quadrant by the contralateral PBN, and the rostromedial quadrant, which receives the weakest input, by the dorsal portion of the nucleus on both sides. These findings suggest that activity in the PBN is relayed to distinct regions of the SC in the form of a highly ordered topographic projection. The adjacent lateral tegmentum (ALT) also projects heavily to the SC, principally on the ipsilateral side. The ALT projection to the ipsilateral SC appears to be organized in a less orderly fashion, and terminates principally in caudal SC, particularly the caudolateral quadrant. No topography was apparent for the contralateral projection.

Animals↗

PubMed1996

Hepatic nitric oxide formation: spin trapping detection in biliary efflux.

A new spin trapping method has been developed to continuously monitor nitric oxide (NO) formation in rat liver in vivo. The method is based on the reaction of NO with iron chelates of N-methylglucamine dithio-carbamate (MGD-Fe), resulting in the formation of room-temperature stable EPR-active nitrosyl complexes of MGD-Fe. Rats were injected with various doses of lipopolysaccharide (LPS) to induce NO synthase activity and were later anesthetized with isoflurane. After cannulation of the bile duct, MGD-Fe was administered by iv injection, and samples of bile were collected for EPR analyses. The EPR spectra of bile from LPS-pretreated rats contained characteristic three-line signals of NO trapped by the MGD-Fe complex, while bile from control rats that were not treated with LPS did not contain similar EPR signals. The detection limit of this method was estimated to be 5 microM. Only weak signals from NO could be detected in plasma or urine under these conditions, suggesting that the biliary NO adducts did not originate in extra-hepatic tissues. The reliability of this method was verified by administering an inhibitor for NO synthase induction, alpha-phenyl-N-t-butylnitrone (PBN), or the NO synthase inhibitor N-nitro-L-arginine methyl ester (L-NAME) to LPS-treated rats. NO detected in bile was significantly decreased by both PBN and L-NAME, which is consistent with results obtained from studies using previously established methods for NO formation.

Animals↗

PubMed1996

Substitute child care at different ages: relationship to social-emotional functioning in preschool.

In a pilot study based on parent and teacher ratings, the number of hours spent in substitute care during the first three years of life correlated with children's levels of behavior problems in preschool. The developmental period from 18 to 24 months was the most sensitive to the use of substitute care, and boys were more negatively affected than girls. The child-adult ratio and setting were not significant factors. Results suggest reconsideration of parental leave policies and direction for future research.

Age Factors↗

PubMed1996

Degradation of DMPO adducts from hydroxyl and 1-hydroxyethyl radicals by rat liver microsomes.

Hydroxyl and 1-hydroxyethyl radical adducts of 5,5-dimethylpyrroline N-oxide (DMPO) were prepared by photolysis, and mechanisms for loss of their EPR signals in rat liver microsomal suspensions were evaluated. Rates of NADPH-dependent EPR signal loss were more rapid in phosphate buffer than in Tris buffer. Addition of superoxide dismutase (SOD) partially protected the adducts when Tris was used as a buffer, but was relatively ineffective in the presence of phosphate. The ferrous iron chelator bathophenan-throlene partially protected the spin adducts in the presence and absence of phosphate, but complete protection was observed when SOD was also added. The spin adducts were unstable in the presence of Fe+2 and K3Fe(CN)6, but Fe+3 alone had little effect on the EPR signals. The data are consistent with two mechanisms for microsomal degradation of DMPO spin adducts under these conditions. Microsomes from superoxide in the presence of oxygen and NADPH, which attacks these DMPO spin adducts directly. The spin adducts are also degraded in the presence of Fe+2, and phosphate stimulates this iron-dependent destruction of DMPO spin adducts.

Animals↗

PubMed1995

Lateral superior olive projections to the inferior colliculus in normal and unilaterally deafened ferrets.

We have examined the projection from the lateral superior olive (LSO) to the inferior colliculus (IC) in the ferret, with particular interest in the laterality of the projection and in the effects of unilateral cochlear removal in infancy. Large or small deposits of the retrograde tracer wheat germ agglutinin-horseradish peroxidase (WGA-HRP) were made in the IC of anesthetized adult ferrets that either were normally hearing or had been unilaterally deafened in infancy (P5 or P25). After 2 days, the ferrets were perfused, and frontal sections of the brainstem were treated with tetramethyl benzidine. For large deposits of WGA-HRP, equal numbers of labelled neurons were found evenly spread through both LSOs. Smaller deposits of WGA-HRP produced four results that contrasted with previous reports on the cat. First, many more labelled neurons were found in the contralateral than in the ipsilateral LSO. Second, the relative number of labelled neurons in each LSO was independent of whether the deposits were in the ventral or in the dorsal IC. Third, the total number of labelled LSO neurons was independent of whether the deposits were in the ventral or in the dorsal IC. Fourth, the proportion of ipsilateral to contralateral labelled neurons was slightly higher in the medial LSO than in the lateral LSO. Ventral IC deposits resulted in more labelled neurons in the medial LSO, and dorsal IC deposits resulted in more labelled neurons in the lateral LSO, as expected. Neonatal cochlear removal did not change any of these results. We conclude that, in the ferret, the organization of the crossed and uncrossed projections from the LSO to the IC differs from that of the cat, and any similarity with the optic chiasm is not obvious.

Animals↗

PubMed1995

Afferent reorganisation within the superior olivary complex of the gerbil: development and induction by neonatal, unilateral cochlear removal.

Cochlear removal in young animals has been shown to produce a variety of degenerative and generative effects within the auditory brainstem. A primary target for axons of neurons in the anteroventral cochlear nucleus (AVCN) is the superior olivary complex (SOC). Following unilateral cochlear removal in neonatal gerbils, AVCN neurons on the side of the removal die, and axons deriving from the AVCN on the unlesioned side produce new endings that innervate previously inappropriate target zones within the ipsilateral medial nucleus of the trapezoid body, both medial superior olives, and the contralateral lateral superior olive. In this study, we have used the anterograde transport of DiI and HRP from the AVCN to relate the formation of these endings to the time course of normal development in the gerbil brainstem. We have also examined the effects of cochlear removal at different ages, and survival to various ages after the removal, to define the time course for these generative phenomena. The results show that, while the major projection pathways from the AVCN to the SOC are in place at the time of birth, further and subtle development of AVCN terminal arbors occurs during the first postnatal week. This overlaps with the time during which cochlear removal produces the formation of exuberant afferents to the SOC from the intact AVCN. The exuberant afferents form through axon sprouting rather than through a suppression of normal, developmental regression. They appear to innervate tonotopically appropriate target regions within the SOC. The formation of the novel afferents begins within 3 days of the removal and appears to be complete within a further 5-7 days. By postnatal day (P) 10, both the normal development of the AVCN to SOC projection and the potential for alteration of that projection by removal of the contralateral cochlea appear to be over. These results suggest that the potential for forming novel projections in the gerbil auditory brainstem is lost before the onset of functional hearing (at P12) and is, therefore, unlikely to result from changes in auditory experience.

Afferent Pathways↗

PubMed1995

Inorganic phosphate promotes redox cycling of iron in liver microsomes: effects on free radical reactions.

The phosphate buffer concentration used in spin trapping experiments with liver microsomes markedly influenced rates of free radical formation from ethanol and dimethylsulfoxide, but not from carbon tetrachloride. Effects of phosphate concentration on ethanol radical formation were abolished by addition of deferoxamine or bathophenanthrolene, indicating that an iron-phosphate complex might be involved. High concentrations of phosphate stimulated rates of microsomal Fe+3 reduction and facilitated the mobilization of microsomal nonheme iron, but had little effect on a variety of microsomal monooxygenase enzyme activities. Although microsomal oxygen utilization and superoxide production were relatively unaffected by phosphate, hydrogen peroxide concentrations were markedly decreased in the presence of high concentrations of phosphate. Taken together, the data suggest that a ferric-phosphate complex may be enzymatically reduced by microsomal enzymes and NADPH. Reoxidation of ferrous ion is nonenzymatically promoted by phosphate and/or H2O2 produced by the microsomes. During the process of reoxidation, one or more oxidizing intermediates may be formed which initiate secondary free radical reactions. Although the reactivity of the intermediate(s) is similar to that of the hydroxyl radical, no spin trapping evidence was obtained to support this assignment.

Animals↗

PubMed1995

Developmentally regulated sensitivity of Trypanosoma brucei brucei to the cytotoxic effects of human high-density lipoprotein.

The bloodstream developmental stages of the protozoan parasite Trypanosoma brucei brucei are lysed by normal human serum. The cytotoxic factor is a minor sub-class of human high-density lipoprotein termed trypanosome lytic factor (TLF). T.b. brucei rapidly develops resistance to TLF when incubated at 26 degrees C under conditions which allow differentiation to the procyclic, insect midgut developmental stage. This in vitro differentiation system allowed us to correlate loss of TLF sensitivity with other parameters of differentiation to the procyclic form. The onset of resistance to TLF occurs within 2 hr after shifting bloodstream forms to differentiation conditions. TLF resistance is correlated with a rapid but transient decrease in protein synthesis by the parasite, is acquired prior to cell division at 26 degrees C, and precedes the loss of variant surface glycoprotein. In addition, we found binding and uptake of TLF by established procyclic trypanosomes was reduced approximately fivefold relative to bloodstream trypanosomes and the TLF binding observed in procyclics was nonspecific. No TLF was bound to the procyclic flagellar pocket membrane and the procyclics failed to endocytose any of the surface-bound TLF. In contrast, bloodstream forms bind TLF via a flagellar pocket-localized protein and bound TLF is taken up by endocytosis. These findings suggest that resistance of procyclic T. b. brucei to TLF-mediated lysis is due to a reduction in the endocytosis of TLF by this developmental stage of the parasite.

Animals↗

PubMed1995

Reconstitution of the trypanolytic factor from components of a subspecies of human high-density lipoproteins.

Trypanosoma brucei brucei is non-infectious to man due to the sensitivity of these parasites to the lytic activity of normal human serum. Apolipoproteins (apo) have been purified, under non-denaturing conditions, from the subclass of human high-density lipoprotein (HDL), termed trypanosome lytic factor (TLF), which is responsible for the cytotoxicity of human serum to T. b. brucei. The TLF apolipoproteins were purified by anion exchange chromatography in the presence of the nonionic detergent octylglucoside and a reconstitution method was developed which allowed the role of the individual apolipoproteins and different lipids to be assessed. The results suggest that the TLF lipids do not have a direct role in lysis but are necessary for the correct assembly of the lytic HDL particle. Apo A-I, apo L-III and apo L-I contribute to lysis in reconstituted particles but individually they are not cytotoxic. Apo A-II was not required in the reconstituted TLF particle for trypanosome lysis. Formation of a lytic HDL particle required apo L-III suggesting its potential role as a toxin. Thermal inactivation of TLF activity correlated with the amount of denatured apo L-I, indicating that apo L-I was involved in lysis of T. b. brucei by native TLF.

Animals↗

PubMed1995

Effects of middle ear pressure on frequency representation in the central auditory system.

Changes in middle ear pressure (MEP) are known to produce an attenuation of sound transmission through the outer and middle ear, but the effects on frequency representation in the auditory system have not previously been studied. This issue is of particular interest because of changes in MEP occurring during episodes of otitis media. We have investigated the effect of changes in MEP on the tuning of neurons in the inferior colliculus (IC) of the gerbil to calibrated tone stimulation of the contralateral, pressurized ear. Both negative and positive non-atmospheric MEP produced an elevation of neural thresholds that was inversely related to IC neuron best frequency (BF). A robust, linear relationship was found between BF at atmospheric MEP (control) and BF at -20 daPa MEP. Higher resolution analysis was performed on a sub-sample of neurons that had particularly stable BFs with repeated, control MEP. For the majority of these neurons, alternation of MEP between control and -20 daPa had no effect on BF. However, a few neurons showed small (up to 5%), significant shifts in BF with -20 daPa MEP. These results are consistent with previous reports of the effects of MEP on spontaneous otoacoustic emissions. We conclude that non-atmospheric MEP acts as a high-pass filter on the input to the cochlea, but does not change the frequency organization of the auditory system to any marked extent.

Acoustic Stimulation↗

PubMed1995

Metabolism of ethanol to 1-hydroxyethyl radicals in vivo: detection with intravenous administration of alpha-(4-pyridyl-1-oxide)-N-t-butylnitrone.

Intravenous administration of the spin-trapping agent alpha-(4-pyridyl-1-oxide)-N-t-butylnitrone (POBN) to anesthetized but otherwise untreated rats was used to test for formation of 1-hydroxyethyl radicals in vivo. The only EPR signals observed in bile samples from rats that had received ethanol but no POBN could be attributed to low concentrations of ascorbyl radical. However, when POBN (700 mg/kg, intravenously) was also administered, a nitroxide with a six-line EPR spectrum was readily detected in bile. This spin adduct was proven to be the 1-hydroxyethyl radical adduct of POBN through injection of [1-13C]ethanol to rats, which resulted in the presence of an adduct with a 12-line EPR spectrum. Comparable results were obtained in experiments with isolated perfused rat livers. 1-Hydroxyethyl radical spin adducts of POBN were readily detectable in bile in the presence of only moderate (10-15 mM) concentrations of alcohol. In these experiments, bile samples were collected into a mixture of dipyridyl and bathocuproine disulfonic acid, and the effectiveness of these chelators to prevent ex vivo signal formation was confirmed experimentally. No EPR signals for nitroxide spin adducts were observed in plasma or perfusate, even though high concentrations of POBN and alcohol were present. Taken together, these data indicate that 1-hydroxyethyl radicals are formed in vivo and can be readily detected in bile when high concentrations of POBN are achieved through intravenous injection.

Animals↗

PubMed1994

Effect of altered neuronal activity on cell size in the medial nucleus of the trapezoid body and ventral cochlear nucleus of the gerbil.

Activity-dependent transneuronal regulation of neuronal soma size has been studied in the medial nucleus of the trapezoid body and ventral cochlear nucleus of adolescent gerbils. Cochlear ablation or tetrodotoxin has been used to eliminate afferent electrical activity in auditory nerve fibers permanently or for 24 or 48 hours. Previous studies have shown that the cross-sectional area of spherical cell somata in the ipsilateral anteroventral cochlear nucleus decreases within 24 hours of electrical activity blockade with tetrodotoxin, which is fully reversible when activity is restored. The present findings extend this work by directly comparing the results of unilateral blockade of auditory nerve action potentials or unilateral cochlear ablation on the size of spherical and globular cell bodies in the ventral cochlear nucleus with changes produced by the same manipulations in third-order cells, principal neurons in the medial nucleus of the trapezoid body. Soma size in both ventral cochlear nucleus cell types decreases reliably by 24 hours after cochlear removal or eighth nerve activity blockade by tetrodotoxin. Soma size of neurons in the contralateral medial nucleus of the trapezoid body decreases 48 hours, but not 24 hours, after either manipulation. When activity in auditory nerve fibers is allowed to resume for 7 days following a 48-hour activity blockade, soma size fully recovers in the medial nucleus of the trapezoid body as well as in ventral cochlear nucleus neurons. We also report that the cross-sectional area of neuronal soma in the medial nucleus of the trapezoid body is larger in lateral regions of medial nucleus of the trapezoid body (low-frequency representation) than in the medial regions of the nucleus (high-frequency representation). We conclude that cell body size changes in brainstem auditory neurons are reversible and that the signals associated with the loss and subsequent recovery of soma size are activity related. However, the delayed effect of activity deprivation in the medial nucleus of the trapezoid body suggests that trophic substances released by afferent axons may contribute to the maintenance of anatomical characteristics.

Action Potentials↗

PubMed1994

Auditory brainstem of the ferret: long survival following cochlear removal progressively changes projections from the cochlear nucleus to the inferior colliculus.

Some effects on auditory brainstem connections of long (1-2.3 years) survival following unilateral cochlear removal in infant and adolescent ferrets were examined by making multiple injections of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) in either the left or the right inferior colliculus (IC). Previous studies have shown that, in normal adult ferrets, about 50 times as many cochlear nucleus (CN) neurons project to the contralateral as to the ipsilateral IC. Right cochlear removal at P25 increased, within 30 days, the number of retrogradely labeled left CN neurons projecting to the left ipsilateral IC by 17% (from n = 235 to n = 275), relative to normals. In this study, longer survival (3 months to 1 year) after right cochlear removal at P25 resulted in larger increases (38-47%; n = 100) in the number of neurons labeled in the left CN after injections of WGA-HRP in the left IC. No change occurred in the number of neurons labeled in the right CN. Taken together, the results of these experiments show that the ratio of the number of labeled neurons in the left CN to that in the right CN increases progressively with survival time out to the maximum time tested (1 year). In contrast to these results, we have previously reported that right cochlear removal at P90 did not change the number of neurons projecting from the left CN to the left IC after 90 days of survival. However, in this study, very long survival (2.3 years) following right cochlear removal at P90 resulted in an increased (51%, from n = 235 to n = 355) number of left CN neurons labeled by WGA-HRP injections into the left IC, relative to normals. The increased number of labeled neurons included neurons throughout each division of the CN and all of the principal morphological types. In a separate series of experiments involving long survival (1-2 years), right cochlear removal at P25 or P40 did not significantly change the number of neurons in either CN retrogradely labeled by injections of WGA-HRP in the right IC, or the ratio between the number of neurons labeled in each CN. Long survival following cochlear removal at P25-P90 did not result in any loss of neurons in the ipsilateral CN or in any shrinkage of CN neurons further than the 10-20% seen at a shorter survival time (90 days).(ABSTRACT TRUNCATED AT 400 WORDS)

Aging↗

PubMed1994

Free-field binaural unmasking in ferrets.

Free-field detection by normal and monaural ferrets of a 500-Hz tone presented over 1 laterally placed loudspeaker and partially masked by narrow-band noise from 2 sources was studied at 2 angular separations of the noise sources (0 degree and 180 degrees). Monaural listening was achieved either by plugging 1 ear canal or removing 1 cochlea. Normal ferrets showed an improvement in detectability of the tone when there was a 180 degrees separation between the noise sources. This unmasking of the tone was abolished in both groups of monaural ferrets, suggesting that the unmasking was due to binaural processing. The development of an animal model demonstrating free-field binaural unmasking, in a species other than humans, will allow investigation into the functional consequences of experimental hearing loss.

Animals↗

PubMed1994

Endocytosis of a cytotoxic human high density lipoprotein results in disruption of acidic intracellular vesicles and subsequent killing of African trypanosomes.

The host range of Trypanosoma brucei brucei is restricted by the cytolytic effects of human serum high-density lipoprotein (HDL). The lytic activity is caused by a minor subclass of human serum HDL called trypanosome lytic factor (TLF). TLF binds in the flagellar pocket to specific TLF-binding sites. Internalization and localization of TLF to a population of endocytic vesicles, and ultimately large lysosome-like vesicles, precedes lysis of T. b. brucei. The membranes of these large vesicles are disrupted by the accumulation of TLF particles. Inhibitor studies with lysosomotropic amines have shown these large vesicles to be acidic in nature and that prevention of their rupture spares the cells from TLF-mediated lysis. Furthermore, leupeptin inhibition suggests that a thioprotease may be involved in the mechanism of TLF-mediated lysis of T. b. brucei. Based on these results, we propose a lytic mechanism involving cell surface binding, endocytosis and lysosomal targeting. This is followed by lysosomal disruption and subsequent autodigestion of the cell.

Acids↗

PubMed1994

Topographic representation of auditory space in the superior colliculus of adult ferrets after monaural deafening in infancy.

1. We have investigated the role of monaural cues provided by the outer ear in the construction of a map of auditory space in the superior colliculus. Single-unit recordings were made from the superior colliculus of adult ferrets that were deprived of binaural inputs by surgically ablating the ipsilateral cochlea on postnatal day 21 or 24. 2. The spatial response properties of auditory units in the deeper layers of this nucleus were studied using white-noise bursts presented under free-field conditions in an anechoic chamber. The thresholds of the units recorded in the monaural ferrets were not significantly different from those recorded in the superior colliculus of normal adult ferrets. In both groups the unit thresholds varied by 30-50 dB in each region of the superior colliculus. 3. In normal and monaural ferrets the elevation tuning tended to be sharper than the azimuth tuning. At sound levels of approximately 10 dB above threshold the auditory units recorded in both groups of animals were tuned to a specific region of space that was restricted in azimuth and elevation. The spatial location at which the maximum response was obtained (auditory best position) varied topographically in azimuth along the rostrocaudal axis of the nucleus and in elevation along the mediolateral axis. 4. The azimuthal distribution of best positions associated with each recording location in the superior colliculus of the monaural ferrets and the alignment between this dimension of the auditory map and that of the visual map in the overlying superficial layers were no different from those found at corresponding near-threshold sound levels in normal ferrets. 5. Elevation spatial selectivity was examined in a smaller sample of units. Although elevation best positions shifted downward from the medial to the lateral side of the nucleus in both normal and monaural ferrets, we found that the topography of the auditory representation and its alignment with the visual representation were statistically different in the two groups of animals. 6. Increasing the sound level does not affect the representation of auditory space in normal ferrets. However, when the stimulus level presented to monoaural ferrets was increased, the receptive fields either expanded so that the responses were no longer tuned to any particular region of space, or the responses remained tuned but exhibited a marked shift in the value of the best position.(ABSTRACT TRUNCATED AT 400 WORDS)

Acoustic Stimulation↗

PubMed1994

Adrenocorticotropic hormone stimulates a transient calcium uptake in rat lymphocytes.

Freshly isolated rat lymphocytes were tested for corticotropin (ACTH)-dependent calcium uptake. Physiological levels of corticotropin (0.01-1 nM) were found to stimulate both an uptake of 45Ca2+ and a rise in cAMP. The calcium uptake was delayed by 2 min after ACTH addition, but was rapid and transient after the onset of uptake. The extent of calcium uptake was dose dependent on the corticotropin concentration and reached a maximum by 1 nM. Several fragments of corticotropin were tested for activity; both full-length 1-39 and a functional truncated form, 1-25, had equivalent effects on 45Ca influx at 1 nM; however, alpha MSH-(1-13), ACTH-(11-24), or a mixture of alpha MSH and ACTH-(11-24) had no effect on 45Ca influx. Extracellular calcium uptake was blocked by the calcium channel blockers lanthanum, diltiazem, nifedipine, and omega-conotoxin. Splenic lymphocytes that express ACTH receptors had ligand-dependent calcium uptake, but thymocytes that lack ACTH receptors had no ligand-dependent calcium uptake. A mouse adrenal cell line, Y-1, showed the same 45Ca uptake kinetics. These findings demonstrate that both lymphocytes and adrenal cells have a functional ACTH-dependent calcium uptake mechanism.

Adrenal Glands↗

PubMed1994

Metabolism of ethanol to 1-hydroxyethyl radicals in rat liver microsomes: comparative studies with three spin trapping agents.

Metabolism of ethanol to 1-hydroxyethyl radicals by rat liver microsomes was studied with three nitrone spin trapping agents (POBN, PBN, and DMPO) under essentially comparable conditions. The data indicate that POBN was the superior spin trapping agent for 1-hydroxyethyl radicals, and that DMPO was least efficient. Addition of deferoxamine completely prevented detection of 1-hydroxyethyl radicals with PBN or DMPO, but caused only 50% decrease in EPR signals when POBN was the spin trap. However, superoxide dismutase only decreased 1-hydroxyethyl radical formation when POBN was the spin trap. Other experiments demonstrated that POBN was the most effective of these nitrones for reduction of Fe(III) in aqueous solutions. Furthermore, 1-hydroxyethyl radical adducts were formed when POBN was added to mixtures of ethanol, phosphate buffer, POBN and FeCl3, but this effect did not occur with either PBN or DMPO. Thus, these data indicate that undesirable effects of POBN on iron chemistry may influence results of spin trapping experiments, and complicate interpretation of the resulting data.

Animals↗