Serial Probe Recognition Task Research Articles

Conjugates of human serum butyrylcholinesterase and nerve agents are behaviorally safe in rhesus macaques

Exogenously administered human serum butyrylcholinesterase (Hu BChE) affords protection by binding to organophosphorus (OP) nerve agents and pesticides in circulation. The resulting Hu BChE-OP conjugate undergoes 'aging' and the conjugate circulates until cleared from the body. Thus, we evaluated the effects of Hu BChE-OP conjugates on the general health and operant behavior of macaques. Rhesus macaques trained to perform a six-item serial probe recognition (SPR) task were administered 30mg/kg of Hu BChE-soman conjugate (n=4) or Hu BChE-VX conjugate (n=4) by intramuscular injection. Performance on the SPR task was evaluated at 60-90min after conjugate administration and daily thereafter for the next 4 weeks. Diazepam (3.2mg/kg), a positive control, was administered 5 weeks after conjugate administration and performance on the SPR task was evaluated as before. Blood collected throughout the study was analyzed for acetylcholinesterase (AChE) and BChE activities. Residual BChE activity of conjugates displayed a similar pharmacokinetic profile as free Hu BChE. Neither of the Hu BChE-OP conjugates produced clear or pronounced degradations in performance on the SPR task. In contrast, diazepam clearly impaired performance on the SPR task on the day of administration in 7 of 8 macaques (and sometimes longer). Taken together, these results suggest that Hu BChE-OP conjugates are safe and provide further support for the development of Hu BChE as a bioscavenger for use in humans.

Characterization of human serum butyrylcholinesterase in rhesus monkeys: Behavioral and physiological effects

The effects of a large dose of human serum butyrylcholinesterase (HuBChE) were evaluated in rhesus monkeys using a serial-probe recognition (SPR) task designed to assess attention and short-term memory. Each monkey received an intravenous injection of 150mg (105,000U or 30mg/kg) of HuBChE 60min prior to testing on the SPR task. Concurrent with the cognitive–behavioral assessment, blood was collected at various time points throughout the study and was analyzed for acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities, anti-BChE antibody production and gross clinical pathology (i.e., complete blood count and clinical chemistry panel). HuBChE revealed a peak blood activity of 227U/ml at 5min after intravenous injection and a mean residence time of approximately 72h. No cognitive–behavioral decrements of any kind in SPR performance and no toxic signs in clinical pathology were detected in any of the blood assays during the 5weeks of observation. Anti-HuBChE antibodies peaked at about 14days after injection, with no concomitant behavioral changes. These results demonstrate the behavioral and physiological safety of HuBChE in rhesus monkeys and support its development as a bioscavenger for the prophylaxis of chemical warfare agent toxicity in humans.

Determination of threshold adverse effect doses of percutaneous VX exposure in African green monkeys

Percutaneous exposure to the chemical warfare nerve agent VX was evaluated in African green monkeys ( n = 9). Doses of VX (7.5–100 μg/kg) were applied to the skin for 60 min and residual agent was quantified (before decontamination) to estimate the absorbed dose. Monkeys were evaluated for the presence or absence of clinical signs of toxicity and blood was sampled periodically (30 min–12 weeks) following exposure to measure the degree of circulating acetylcholinesterase (AChE) inhibition. Monkeys were also evaluated for behavioral changes from VX exposure using a serial probe recognition (SPR) task. The lowest observable adverse effect level (LOAEL) for the production of major clinical signs was determined to be 42.22 μg/kg (absorbed dose estimate = 17.36 μg/kg) and the LOAEL for AChE inhibition was 13.33 μg/kg (absorbed dose estimate = 6.53 μg/kg). Behavioral performance was unaffected at doses that, while producing substantial AChE inhibition, did not produce clinical signs. VX represents a substantial threat as a contact hazard and these results complement previous studies using the percutaneous route of exposure with VX and extend the findings to a non-human primate species.

ORIGINAL ARTICLE: Evaluation of miosis, behavior and cholinesterase inhibition from low‐level, whole‐body vapor exposure to soman in African green monkeys (Chlorocebus sabeus)*

Relatively little is known about the effects of very low-level exposures to nerve agents where few signs or symptoms are present. African green monkeys (Chlorocebus sabeus) (n = 8) were exposed for 10 min, whole-body, to a single concentration of soman (0.028-0.891 mg/m³). EC₅₀ values for miosis were determined to be 0.055 mg/m³ and 0.132 mg/m³ when defined as a 50 percent reduction in pupil area and diameter, respectively. In general, performance on a serial probe recognition task remained unchanged at lower concentrations, but responding was suppressed at the largest concentration tested. Soman produced concentration-dependent inhibition of acetylcholinesterase activity and, to a lesser extent, butyrylcholinesterase activity. These results characterize threshold soman exposure concentrations that produce miosis in the absence of other overt signs of toxicity and extend previous studies indicating that miosis is a valuable early indicator for the detection of soman vapor exposure.

Systemic administration of the potential countermeasure huperzine reversibly inhibits central and peripheral acetylcholinesterase activity without adverse cognitive–behavioral effects

Huperzine A is potentially superior to pyridostigmine bromide as a pretreatment for nerve agent intoxication because it inhibits acetylcholinesterase both peripherally and centrally, unlike pyridostigmine, which acts only peripherally. Using rhesus monkeys, we evaluated the time course of acetylcholinesterase and butyrylcholinesterase inhibition following four different doses of -(-)huperzine A: 5, 10, 20, and 40 μg/kg. Acetylcholinesterase inhibition peaked 30 min after intramuscular injection and varied dose dependently, ranging from about 30% to 75%. Subsequently, cognitive–behavioral functioning was also evaluated at each dose of huperzine A using a six-item serial-probe recognition task that assessed attention, motivation, and working memory. Huperzine did not impair performance, but physostigmine did. The results demonstrate that huperzine A can selectively and reversibly inhibit acetylcholinesterase without cognitive–behavioral side effects, thus warranting further study.

Enhanced recognition of emotional stimuli in the chimpanzee (Pan troglodytes)

Humans often have a better memory of emotional events than neutral events. From the comparative-cognitive perspective, we explored the enhancement of recognition memory by emotion in chimpanzees (Pan troglodytes) using a serial probe recognition task. In this task, we sequentially presented a list of pictures to subjects and then tested their recognition of specific pictures from within the list. We selected pictures of aggressive chimpanzees as emotional stimuli and less tensed, relaxed chimpanzees as neutral stimuli. In Experiment 1, we gave four-item lists to two young chimpanzees; one showed significantly greater recognition of pictures depicting aggressive chimpanzees than neutral ones. In Experiment 2, this chimpanzee was further tested using a recognition task with eight-item lists. The subject again showed better recognition of emotional stimuli than neutral. Furthermore, the presence of an emotional stimulus in the list also facilitated recognition of the neutral item immediately following it. Overall, although only one of the two chimpanzees showed enhanced recognition memory by emotional stimuli, this is the first demonstration of such a response in the chimpanzee. The findings are discussed in comparison with those of human studies.

Evaluation of cognitive and biochemical effects of low-level exposure to sarin in rhesus and African green monkeys

We investigated the potential of low-level exposures to the chemical warfare nerve agent, sarin, to produce adverse effects. Rhesus ( Macaca mulatta) and African green monkeys ( Chlorocebus acthiops) were trained on a serial probe recognition (SPR) task before IM administration of a low-level concentration (5.87 μg/kg or 2.93 μg/kg) of sarin. Blood was sampled before agent administration and at various times following administration. Sarin administration did not disrupt performance on the SPR task in either species. Major dependent measures characterizing performance (accuracy, number of completed trials per session, average choice response time) were largely unaffected on the day sarin was administered as well as on subsequent testing sessions occurring over several weeks following administration. Analyses of red blood cell (RBC) and plasma samples revealed that sarin administration produced a substantial degree of inhibition of circulating acetylcholinesterase (AChE) in RBC fractions and butyrylcholinesterase (BChE) in plasma fractions, which only slowly recovered. In this regard, AChE activity was inhibited to a greater extent than BChE activity. Blood samples were also evaluated for regenerated sarin, which was found in RBC and plasma fractions in both species and showed orderly elimination functions. More sarin was regenerated from RBC fractions than from plasma fractions. Elimination of regenerated sarin was much slower in RBC than plasma and exceeded the expected time of AChE aging, suggesting the presence of additional sarin binding sites. In general, effects were similar in both species. Taken together, our results show that while the concentrations of sarin administered were clearly biochemically active, they were below those that are required to produce a disruption of behavioral performance.

Serial-probe recognition in rhesus macaques: Effects of midazolam

A serial-probe recognition task was used to assess the effects of midazolam on visual attention and short-term memory in three rhesus monkeys. On each trial, six unique alphanumeric sample stimuli (list items) were presented sequentially followed by a choice period. Choosing the ‘probe’ stimulus was correct if the probe matched one of the list items; otherwise, choosing the ‘default’ stimulus (a white square) was correct. Behavior was examined under a range of doses of midazolam (0.065, 0.13, 0.26, and 0.52 mg/kg IM). Midazolam did not significantly reduce choice accuracy or change the shape of the serial position function and did not significantly reduce choice responding. However, choice reaction time was significantly increased by the two highest doses of midazolam. Responding directed at the sample stimuli was reduced at the two highest doses of midazolam. Furthermore, 0.52 mg/kg midazolam significantly increased sample-stimulus reaction time at all six serial positions. Overall, these data suggest that the two highest doses of midazolam tested increase reaction time, but do not directly impair short-term visual recognition memory. This is noteworthy because such doses appear capable of protecting against nerve agent-induced seizures.

P.4.017 Protection against organophosphates inducedirreversible acetylcholinesterase inhibition by concomitant administration of donepezil and pyridostygmine

The possibility that nerve agents will be used on the battlefield is real. The traditional therapy against nerve agent exposure consists of pyridostigmine pretreatment and atropine-pralidoxime chloride therapy administered after nerve agent exposure. This therapy regimen is extremely effective in preventing mortality in laboratory animals exposed to multilethal concentrations of nerve agent, yet these animals often display convulsions, brain damage, and behavioral incapacitation. We report here that the addition of diazepam to the traditional therapy for nerve agent (soman) exposure not only decreases the incidence of convulsions, but also attenuates the cognitive impairments of rhesus monkeys trained on a Serial Probe Recognition (SPR) task. Monkeys which received diazepam treatment required only 6 days before their performance on the SPR task returned to presoman exposure levels, compared to nondiazepam-treated monkeys which required 15 days. Moreover, only 1 out of the 5 monkeys which received diazepam treatment suffered tonic-clonic convulsions; in contrast all 5 monkeys which did not receive diazepam treatment experienced severe convulsive episodes. These results suggest that diazepam would be an excellent adjunct to traditional nerve agent therapy to facilitate behavioral recovery from nerve agent intoxication that might be encountered by US military personnel on the battlefield or accidental organophosphate poisoning encountered in industrial or agricultural accidents.

Immediate and delayed stimulus repetitions evoke different ERPs in a serial‐probe recognition task

In this study we examined whether event‐related potentials (ERPs) associated with stimulus repetition and recognition in a serial‐probe recognition task were comparable to ERPs in other tasks that are more typically used to investigate old/new ERP effects. The experiment consisted of 320 trials in which a recognition probe followed a four‐item memory set; 160 trials consisted of images depicting common objects that were easy to label (EL task), and 160 trials consisted of images depicting abstract patterns that were difficult to label (DL task). Nineteen participants indicated whether a probe that followed each memory set was or was not presented in the memory set. Half of the probes matched, and half did not match, an item in the preceding memory set. ERPs appeared to reflect two processes—one that differentiated between recently presented stimuli and other stimuli and another that distinguished between repeated stimuli and new stimuli. ERPs to recent probes were more positive than ERPs to other probes in the EL and DL tasks. ERPs to match (old) probes were more positive than ERPs to nonmatch (new) probes only in the EL task.

Immediate and delayed stimulus repetitions evoke different ERPs in a serial-probe recognition task

In this study we examined whether event-related potentials (ERPs) associated with stimulus repetition and recognition in a serial-probe recognition task were comparable to ERPs in other tasks that are more typically used to investigate old/new ERP effects. The experiment consisted of 320 trials in which a recognition probe followed a four-item memory set; 160 trials consisted of images depicting common objects that were easy to label (EL task), and 160 trials consisted of images depicting abstract patterns that were difficult to label (DL task). Nineteen participants indicated whether a probe that followed each memory set was or was not presented in the memory set. Half of the probes matched, and half did not match, an item in the preceding memory set. ERPs appeared to reflect two processes--one that differentiated between recently presented stimuli and other stimuli and another that distinguished between repeated stimuli and new stimuli. ERPs to recent probes were more positive than ERPs to other probes in the EL and DL tasks. ERPs to match (old) probes were more positive than ERPs to nonmatch (new) probes only in the EL task.

Behavioral and Immunological Effects of Exogenous Butyrylcholinesterase in Rhesus Monkeys

Although conventional therapies prevent organophosphate (OP) lethality, laboratory animals exposed to such treatments typically display behavioral incapacitation. Pretreatment with purified exogenous human or equine serum butyrylcholinesterase (Eq-BuChE), conversely, has effectively prevented OP lethality in rats and rhesus monkeys, without producing the adverse side effects associated with conventional treatments. In monkeys, however, using a commercial preparation of Eq-BuChE has been reported to incapacitate responding. In the present study, repeated administration of commercially prepared Eq-BuChE had no systematic effect on behavior in rhesus monkeys as measured by a six-item serial probe recognition task, despite 7- to 18-fold increases in baseline BuChE levels in blood. Antibody production induced by the enzyme was slight after the first injection and more pronounced following the second injection. The lack of behavioral effects, the relatively long in vivo half-life, and the previously demonstrated efficacy of BuChE as a biological scavenger for highly toxic OPs make BuChE potentially more effective than current treatment regimens for OP toxicity.

Auditory list memory and interference processes in monkeys.

Memory of 2 rhesus monkeys (Macaca mulatta) was tested in a serial probe recognition task with lists of 4 natural or environmental sounds, different retention intervals, and different manipulations of interference. At short retention intervals, increasing the separation of list items reduced the primacy effect and produced a recency effect. Similar results were shown by increasing interference across lists through item repetitions or making the first 2 list items high-interference items. These results indicated that decreasing first-item performance reduced proactive interference on memory of the last list items. At long (20 s) retention intervals, making the last list items of high interference reduced the recency effect, reduced retroactive interference, and produced a primacy effect. Taken together, interference plays a role in determining the primacy and recency effects of the serial-position function.

Primacy and recency effects in rhesus monkeys (Macaca mulatta) using a serial probe recognition task. III. A developmental analysis.

In children, the recency effect emerges prior to the primacy effect. To determine whether this dissociation is also seen in nonhuman primates, we evaluated the development of the primacy and recency effect in 3 young adult (35 months) and 4 adolescent (21 months) male rhesus monkeys (Macaca mulatta) using a six-item serial probe recognition (SPR) task. As predicted, the young adult monkeys displayed both effects, while the adolescent monkeys only displayed the recency effect. Not until after 26 months of training on the SPR task did the adolescent monkeys exhibit both the primacy and recency effect. Interference and strategy differences are discussed in terms of the results along with an interpretation of Rudy's (1992) configural association theory of cognitive development. Additional possible explanations for this developmental dissociation include the delayed maturation of the neocortical, hippocampal, and/or cholinergic systems, the latter two having been shown to be important in the expression of the primacy but not the recency effect.

Auditory List Memory in Rhesus Monkeys

Auditory memory of 2 rhesus monkeys was tested in a serial probe recognition task. Lists of four environmental or natural sounds were followed by a retention interval and a test. The test matched one of the list items on half of the trials. The retention interval was varied across sessions. Six experiments showed similar results and changes in the serial position function. At short retention intervals, there was good memory for first list items (primacy effect) and poor memory for last list items. At intermediate retention intervals, memory improved for last list items (recency effect). At long retention intervals (20 s and 30 s), the recency effect was strong, and the primacy effect had dissipated. These auditory primacy and recency effects and their changes with retention interval were opposite to those for visual memory. Implications for processes and mechanisms of memory are discussed.

Primacy and recency effects in rhesus monkeys (Macaca mulatta) using a serial probe recognition task: II. Effects of atropine sulfate.

Primacy and recency effects in rhesus monkeys (Macaca mulatta) using a serial probe recognition task: II. Effects of atropine sulfate.

Primacy and recency effects in rhesus monkeys (Macaca mulatta) using a serial probe recognition task: II. Effects of atropine sulfate.

Nonhuman primates display both a primacy and a recency effect when trained on a 6-item serial probe recognition task. The author has previously shown that in the rhesus monkey, diazepam (3.2 mg/kg im) interferes with the memory processes that mediate the recency effect without affecting those memory processes involved in the primacy effect (C. A. Castro, 1995). This study assessed the effects of atropine sulfate (0.2, 0.3, and 0.4 mg/kg im) on the primacy and recency effects in these same monkeys. Opposite the effects of diazepam, atropine disrupted the primacy component of the serial position curve and had no measurable effect on the recency component. In addition, the 2 highest doses of atropine disrupted accuracy on the nonmatching probe trials, whereas all 3 doses of atropine resulted in increased response latencies. These reports indicate that the primacy and recency effects in the nonhuman primate can be pharmacologically dissociated.

Primacy and recency effects in rhesus monkeys (Macaca mulatta) using a serial probe recognition task. I. Effects of diazepam.

In this study, we evaluated the effects of diazepam (0.2, 0.8, 1.6 and 3.2 mg/kg; IM) on the primacy and recency memory effects in four rhesus monkeys trained on a six-item serial probe recognition (SPR) task. Only the highest dose of diazepam (3.2 mg/kg) consistently affected the shape of the monkeys' serial position curves. Accuracy on the probe trials was disrupted for list items which occurred in the middle portion and recency memory component of the serial position curve, without affecting the primacy component. Diazepam, however, also produced several nonspecific effects on SPR performance. Both the 1.6 and 3.2 mg/kg diazepam doses disrupted accuracy on the non-matching probe trials, and the 3.2 mg/kg dose caused an increase in response latencies which were unrelated to any one component of the serial position curve. This is the first demonstration in nonhuman primates showing that the primacy and recency memory effects can be differentiated on the basis of diazepam dose.

Role of the forebrain commissures in bihemispheric mnemonic integration in macaques

A serial probe recognition task was used to examine the interhemispheric exchange of visual data in macaques. Each block of trials began with the memorization of one to six visual target images. The monkeys then had to determine, in tests that followed immediately, whether probe images were or were not members of the learned target set. Previous work with both humans and macaques has shown that the time required for the evaluation of probes generally increases, while response accuracy decreases, as a function of the number of targets, the "memory load". By testing animals with bisected optic chiasm, it was possible to direct visual information to only one hemisphere at a time, simply by occluding the opposite eye. In this fashion, the quality of intrahemispheric evaluations (in which a monocular probe was a match for a target previously viewed through the same eye) was compared with that of interhemispheric evaluations (in which a probe was a match for a target previously designated through the opposite eye). A key question was whether division of the target list between the hemispheres modified the relationships between reaction time, response accuracy, and memory load. Provided that either the anterior commissure or the splenium of the corpus callosum was intact, interhemispheric processing was only subtly less efficient than intrahemispheric processing. The ability to perform interhemispheric evaluations was selectively and completely disrupted if all forebrain commissural fibers were transected. In this latter split-brain condition, the time required for probe evaluations was, as expected, determined solely by the number of target items memorized by the probed hemisphere. Accuracy, however, was always a function of the total memory load, regardless of the distribution of targets between the hemispheres. This implies, first, that accuracy and latency do not reflect identical mnemonic factors, as frequently held, and second, that in mnemonic processing, the two hemispheres draw upon a unified, shared resource, probably allocated by the intact brainstem.

Behavioral decrements persist in rhesus monkeys trained on a serial probe recognition task despite protection against soman lethality by butyrylcholinesterase

Recently, it has been demonstrated that an exogenously administered enzyme such as butyrylcholinesterase (BuChE) can prevent death in rhesus monkeys exposed to multiple-lethal doses of the acetylcholinesterase inhibitor soman when the enzyme is given prior to soman exposure (3). We report that despite BuChE protecting against soman-induced lethality, behavioral effects are seen in these monkeys which last for at least 6 days as measured by performance on a serial probe recognition (SPR) task. Analyses of the serial position curves showed that performance was lower on the probe trials when the probe items were from the middle of the list than when the probe items were from the beginning or end of the list which were unaffected. BuChE given alone also produced behavioral effects, causing all animals not to respond on the probe trials until 8 h following BuChE administration. Taken together, these findings suggest that the BuChE is not completely binding all of the soman and that a concentration of soman which is capable of causing behavioral effects may be entering the CNS.