Encoding and retrieval are differentially processed by the anterior cingulate and prelimbic cortices: a study based on trace eyeblink conditioning in the rabbit.

A growing body of literature suggests that structures along the midline of the prefrontal cortex (mPFC), including Brodmann's area 32 (prelimbic cortex) and area 24 (anterior cingulate cortex) in the rabbit play a role in retrieval of learned information. The present studies compared the effects of post-training lesions produced either immediately or 1-week following learning, to either prelimbic (area 32) or anterior cingulate (area 24) cortex on trace eyeblink (EB) conditioning. Further, because recent evidence suggests that the mPFC may play an even greater role in learning and memory when emotional arousal is low, these studies compared the effects of lesions in groups conditioned with either a relatively low-arousal corneal airpuff, or a more aversive periorbital eyeshock unconditioned stimulus (US). A total of six groups were tested, which received selective ibotenic acid or "sham" control lesions to either area 32 or 24, immediately or 1-week following asymptotic learning, and conditioned with an eyeshock US or an airpuff US. Results showed that the greatest lesion deficits were found when conditioning with the less aversive airpuff US. Further, lesions produced to area 32 one-week, but not immediately following learning, caused significant deficits in performance, while lesions produced to area 24 immediately, but not 1-week following learning, caused significant deficits in performance. These findings add to the body of evidence which shows that area 32 of the mPFC regulates retrieval, but not acquisition or storage of information, while area 24 mediates a less specific reacquisition process, but not permanent storage or retrieval of information during relearning of memories abolished by mPFC damage. These findings were, however, specific to those experiments in which the relatively non-aversive airpuff was the US.

Where does foodborne illness happen-in the home, at foodservice, or elsewhere-and does it matter?

Foodservice professionals, politicians, and the media are often cited making claims as to which locations most often expose consumers to foodborne pathogens. Many times, it is implied that most foodborne illnesses originate from food consumed where dishes are prepared to order, such as restaurants or in private homes. The manner in which the question is posed and answered frequently reveals a speculative bias that either favors homemade or foodservice meals as the most common source of foodborne pathogens. Many answers have little or no scientific grounding, while others use data compiled by passive surveillance systems. Current surveillance systems focus on the place where food is consumed rather than the point where food is contaminated. Rather than focusing on the location of consumption-and blaming consumers and others-analysis of the steps leading to foodborne illness should center on the causes of contamination in a complex farm-to-fork food safety system.

Prefrontal control of trace eyeblink conditioning in rabbits: role in retrieval of the CR?

Rabbits (Oryctolagus cuniculus) were trained on a trace eyeblink (EB) conditioning task to a criterion of 10 consecutive EB conditioned responses (CRs). One week later, ibotenic acid or sham lesions were made in the mPFC centered on the prelimbic region (Brodmann's area 32) or the cingulate cortex (Brodmann's area 24). Following a 1-week postoperative recovery period, all animals were retrained for 4 consecutive days using the same parameters as during acquisition, given 1 week off, and retrained for another 4 days. Mean EB conditioning deficits in the group with area 32 lesions occurred on the first and second days of each retraining period. However, by the third and fourth days of retraining, these lesioned animals were performing at a level comparable to that of the sham group. Lesions of area 24 did not produce deficits at either retesting period. These findings were interpreted to indicate that area 32, but not area 24, is involved in retrieval processes, rather than consolidation or storage, in that the animals were impaired at both retesting times, but were able to relearn the task.

Learning and memory impairment in PTSD: relationship to depression.

Memory function was studied in combat veterans with posttraumatic stress disorder (PTSD), combat veterans without PTSD, and noncombat veterans. The Vocabulary and Digit Span subtests of the WAIS and Logical Memory (LMS) and Verbal Paired Associates (VPAS) subtests of the Wechsler Memory Scale III were administered. Combat veterans with PTSD showed impaired memory on the LMS and VPAS compared to combat veterans without PTSD or noncombat veterans. Veterans with PTSD also showed lower WAIS Vocabulary subtest scores--but not digit span subtest scores--than combat veterans without PTSD or noncombat veterans. Medication status, co-morbid diagnosis, and age all failed to account for these memory differences, but when self-assessed depression--as measured by the Zung Self-Rating Depression Scale--or anxiety--as measured by the Spielberger State-Trait Anxiety Scale--was statistically removed, group differences on these memory measures were no longer significant. However, using a stepwise regression procedure, in which both anxiety and depression were employed to predict the LMS and VPAS scores, only the Zung scale reliably predicated performance. The present results, showing that PTSD is associated with general learning and memory impairments, is an important finding, but the specific effects of depression as a mediator of these deficits should be further studied.

Discriminative delay Pavlovian eyeblink conditioning in veterans with and without posttraumatic stress disorder.

BACKGROUND: Impaired eyeblink (EB) classical conditioning using a delay paradigm has previously been shown in combat veterans, as well as in a group of depressed adults, compared to normal individuals. Significant deficits in immediate memory (IM) in combat PTSD+ veterans, compared to normal controls, have also been previously shown, but these differences became non-significant after controlling for level of self-reported depression. Furthermore, EB conditioning has been shown to be significantly correlated with heart rate variability (HRV) in normal adults. The present study examined how depression (self-reported), IM, and resting HRV are related to discriminative delay classical EB conditioning in veterans with and without PTSD. METHOD: Three groups of subjects (combat PTSD+, combat PTSD-, and non-combat PTSD-) were assessed for self-report of depression and anxiety, as well as IM and HRV. Subjects received a single session of discriminative EB classical conditioning in which the conditioned stimulus (CS) was a light signal (either red or green) compounded with a tone. On CS+ trials, the light-tone compound stimulus co-terminated with a corneal airpuff (unconditioned stimulus, US), thus producing a delay paradigm. On CS- trials the appropriate light-tone stimulus was presented but not followed by the airpuff US. EB amplitude and frequency were recorded. RESULTS: PTSD+ subjects had greater self-reported depression and anxiety scores than the two control groups, as well as lower scores on a measure of IM. However, the IM difference was not significant after the effects of self-reported depression and anxiety were controlled. EB CR amplitude was significantly greater to CS+ than CS- for all three groups. EB amplitude to both the US (airpuff) and the CS+ declined over trials, but was significantly lower in the combat PTSD+ group compared to the combined PTSD- groups. Subjects who reached an EB CR acquisition criterion had significantly greater scores on IM than those who did not reach criterion. Factor analysis of the entire data set revealed four factors corresponding to (1) self-reported depression and anxiety, (2) IM, (3) HRV, and (4) EB amplitude. EB frequency was significantly predicted by IM and HRV. CONCLUSIONS: These data extend our previous results by showing deficits in EB conditioning among combat PTSD+ veterans that were associated with lower IM and resting HRV, but were not associated with self-report of depression.

Ibotenic acid lesions to ventrolateral thalamic nuclei disrupts trace and delay eyeblink conditioning in rabbits.

Intact cerebellar structures (i.e., deep nuclei and perhaps cortex) are essential for acquisition of both simple delay and trace eyeblink (EB) conditioning. However, successful trace conditioning also requires intact cortico-limbic structures (i.e., hippocampus, medial thalamus, and medial prefrontal cortex, mPFC). A direct connection between the cerebellum and ventrolateral thalamic nuclei (VLTN) has been demonstrated in several species. Since VLTN projects to both premotor and prefrontal cortex, it may be an essential link in a cerebellar-thalamic-prefrontal circuit that provides the CNS substrate for acquisition of the trace EB CR. The current studies thus assessed the role of the VLTN on trace EB conditioning in New Zealand albino rabbits. We first verified afferent connections to the mPFC (Brodmann's area 32) from the VLTN, by injecting the retrograde tracer Flourogold(c) into area 32. Strong labeling in VLTN from terminal projections to mPFC were found. We next assessed the role of VLTN in trace eyeblink conditioning in animals that received either sham or ibotenic acid VLTN lesions. EB conditioning began with 10 consecutive daily sessions of trace conditioning, followed immediately by 4 days of extinction, and then 4 days of delay conditioning. VLTN lesions significantly impaired acquisition of both trace and delay conditioning, and impaired extinction. These findings, thus confirm the importance of the VLTN in a postulated cerebellar-thalamic-prefrontal circuit that underlies successful trace, as well as delay EB conditioning.

Combat veterans show normal discrimination during differential trace eyeblink conditioning, but increased responsivity to the conditioned and unconditioned stimulus.

The question addressed in the present study was whether post-traumatic stress disorder (PTSD) results in associative learning impairments. To answer this question, differential trace eyeblink (EB) conditioning was studied in combat veterans with PTSD, combat veterans without PTSD, and non-combat veterans without PTSD. Veterans with PTSD showed normal EB discrimination, suggesting that associative learning is not impaired by PTSD. Veterans with PTSD also showed normal extinction. However, subjects with PTSD showed more EB conditioned responses (CRs), as well as increased CR amplitude. Increased response amplitude to the airpuff unconditioned stimulus presented alone (viz. the unconditioned response), as well as to the airpuff on CS+ trials during conditioning also occurred in the subjects with PTSD. These findings suggest increased reactivity in combat veterans with PTSD, compared to those without PTSD, but such heightened reactivity does not affect somatomotor associative learning.

Prefrontal control of trace versus delay eyeblink conditioning: role of the unconditioned stimulus in rabbits (Oryctolagus cuniculus).

The conditioned eyeblink (EB) response was studied with trace conditioning procedures in rabbits (Oryctolagus cuniculus) with lesions to the medial prefrontal cortex (mPFC) or sham lesions. Three experiments were performed in which either periorbital shock or a corneal airpuff served as the unconditioned stimulus (US) in separate groups of sham or mPFC-lesioned rabbits. Acquisition of the EB conditioned response (CR) was faster and reached a higher asymptote with the eyeshock US than with the airpuff US. However, mPFC lesion-induced trace conditioning deficits were obtained only in the groups that received the airpuff US. All rabbits showed normal delay conditioning and extinction. These results suggest that mPFC mediates trace EB conditioning when emotional arousal is low. However, in circumstances when emotional arousal may be high (i.e., during exposure to aversive periorbital shock), other structures (such as amygdala) may be activated to permit learning even in the absence of input from mPFC.

Nicotine modulation of cytokine induction by LPS-stimulated human monocytes and coronary artery endothelial cells.

Nicotine, the major addictive component of tobacco, is an immunomodulator that impacts on many cells, including immune cells involved in inflammatory processes. Nicotine also induces oxidative damage to the vascular endothelium and accentuates lipid peroxidation, resulting in vascular cell dysfunction. Furthermore, vascular endothelial cells produce growth factors, such as cytokines and chemokines capable of stimulating and recruiting immune cells to atheromatous lesions. In addition, bacterial products including lipopolysaccharides (LPS), a major component of Gram negative bacterial cell walls, activate gene expression resulting in inflammatory cytokine production causing further damage to the vasculature. In the present study, the combined effects of nicotine and bacterial LPS on the expression of IL-6, IL-8, GRO-alpha and MCP-1 in cell lines of human coronary artery endothelial cells (HCAEC) and pulmonary monocytes (THP-1) were examined by quantitative real-time PCR and ELISA. Results showed that nicotine suppressed the LPS induced production of IL-6 and IL-8 in both cell lines. Since cytokines which alter homeostasis of both vascular endothelial and immune cells are critical for the atherogenic process, further studies are warranted to examine in detail the role of nicotine in terms of effects on inflammatory reactions, including those induced by bacterial infection.

Single unit activity in the medial prefrontal cortex during Pavlovian heart rate conditioning: Effects of peripheral autonomic blockade.

Electrical activity was recorded from single neurons in the medial prefrontal cortex of rabbits during differential Pavlovian heart rate (HR) conditioning. A heterogeneous population of cells were found, some of which showed CS-evoked increases and others CS-evoked decreases in discharge, while some cells were biphasic. A subset of cells also showed trial-related changes in discharge that were related to acquisition of the HR discrimination between the reinforced CS+ and non-reinforced CS-. Administration of the peripheral cholinergic antagonist, methylscopolamine, and the andrenergic antagonist, atenolol, either increased or decreased maintained baseline activity of many cells, but had little or no effect on the CS-evoked activity of these cells. Waveform changes also did not result from administration of these drugs. This finding suggests that CS-evoked mPFC activity is not being driven by cardiac afferent input to CNS cardiac control centers. Previous studies have shown that ibotenic acid lesions of this area greatly decreases the magnitude of decelerative heart rate conditioned responses; the latter finding, plus the results of the present study, suggest that processing of CS/US contingencies by the prefrontal cortex contributes to the acquisition of autonomic changes during Pavlovian conditioning.

Medial prefrontal lesions and Pavlovian eyeblink and heart rate conditioning: effects of partial reinforcement on delay and trace conditioning in rabbits (Oryctolagus cuniculus).

Effects of continuous (100%) versus partial (25%) reinforcement were studied on Pavlovian delay and trace eyeblink conditioning in rabbits (Oryctolagus cuniculus) with either lesions to the medial prefrontal cortex (mPFC) or sham lesions. Concomitant heart rate changes evoked by the conditioned stimulus were also assessed. Partial reinforcement retarded eyeblink conditioning in both the trace and delay paradigm, but this impairment was greater during trace conditioning and in rabbits with mPFC lesions. Accompanying conditioned stimulus-evoked heart rate slowing was attenuated under all conditions by the mPFC lesions, although this result was not always statistically significant.

The role of claustrum in Pavlovian heart rate conditioning in the rabbit (Oryctolagus cuniculus): anatomical, electrophysiological, and lesion studies.

The role of the claustrum in Pavlovian heart rate (HR) conditioning was studied in the rabbit (Oryctolagus cuniculus) by (a) mapping claustral projections to the prefrontal cortex (PFC), (b) recording claustral single-unit discharge to sensory stimulation and conditioning stimuli during HR conditioning, and (c) assessing the effects of claustral damage on HR conditioning. Contralateral and ipsilateral claustral projections to the PFC were found. Claustral cells responded to nonsignal stimulation with increased discharge and also showed conditioned stimulus-evoked increases in discharge during Pavlovian HR conditioning. Moreover, claustral lesions diminished the magnitude of the HR-conditioned response without affecting the cardiac-orienting response to the conditioned stimulus or the cardiac-unconditioning response to the unconditioned stimulus, suggesting a role for the claustrum in associative learning.

Expressions of hepatic genes, especially IGF-binding protein-1, correlating with serum corticosterone in microarray analysis.

Microarray technology was evaluated for usefulness in assessing relationships between serum corticosterone and hepatic gene expression. Nine pairs of female Swiss mice were chosen to provide a wide range of serum corticosterone ratios; cDNA microarray analysis (approximately 8000 genes) was performed on their livers. A statistical method based on calculation of 99% confidence intervals discovered 32 genes which varied significantly among the livers. Five of these ratios correlated significantly with serum corticosterone ratio, including tyrosine aminotransferase, stress-induced protein, pleiotropic regulator 1 and insulin-like growth factor-binding protein-1; the latter has a potential role in cancer development. Secondly, linear regression of gene expression vs corticosterone ratios was screened for those with r> or =0.8 (P<0.01), yielding 141 genes, including some known to be corticosterone regulated and others of interest as possible glucocorticoid targets. Half of these significant correlations involved data sets where no microarray ratio exceeded +/- 1.5. These results showed that microarray may be used to survey tissues for changes in gene expression related to serum hormones, and that even small changes in expression can be of statistical significance in a study with adequate numbers of replicate samples.

Pavlovian eyeblink conditioning in combat veterans with and without post-traumatic stress disorder.

Several recent studies have investigated relationships between post-traumatic stress disorder (PTSD) and learning and memory problems. These reports have found in general that not only does PTSD affect trauma-related memories, but when patients with PTSD are compared with similar trauma patients without PTSD, general memory impairments have been found. The present paper reports a study in which associative learning, using Pavlovian eyeblink conditioning, was investigated in combat veterans with and without chronic PTSD, using interstimulus intervals of 500 and 1000 msec in two separate experiments. Although several recent reports suggest that larger-magnitude autonomic conditioned responses occur in patients with PTSD during Pavlovian conditioning, the present study found evidence of impaired Pavlovian eyeblink conditioning in combat veterans with and without PTSD, compared to non-combat veterans. Although these data suggest that combat leads to an impaired associative learning process regardless of whether PTSD is apparent, a group of community-dwelling combat veterans not under medical treatment showed normal conditioning, suggesting that variables other than prior combat must also be involved.

Posttraining prefrontal lesions impair jaw movement conditioning performance, but have no effect on accompanying heart rate changes.

This experiment examined the role of the medial prefrontal cortex (mPFC) in regulating learned autonomic and somatomotor responses in rabbits using appetitive Pavlovian conditioning. Interstimulus interval (ISI) duration [i.e., the time between the onset of the conditioned stimulus (CS) and unconditioned stimulus (US)] was manipulated in order to determine whether ISI duration was related to the heart rate (HR) responses obtained during conditioning. Two groups received either a 1- or a 4-s ISI, with a tone as the CS and an intraoral pulse of water as the US. Another two groups received explicitly unpaired presentations of either the 1- or 4-s tone CS and water US. Few conditioned jaw movement (JM) or HR conditioned responses (CRs) were observed in the unpaired conditions. Significant JM conditioning was, however, elicited by the paired conditions, especially to the 4-s ISI. Consistent CS-evoked HR accelerations were observed in both ISI conditions. After five sessions of training, the mPFC was lesioned in half the animals. A separate group of paired animals received sham lesions. After surgical recovery, all animals received 3 days of postoperative training. During the first postoperative training session, JM CRs significantly declined in both groups with mPFC lesions in comparison to the groups with sham lesions. The mPFC lesions, however, did not affect the CS-evoked cardiac accelerations, which again occurred during postoperative training.

Heart rate changes accompanying jaw movement Pavlovian conditioning in rabbits: concomitant blood pressure adjustments and effects of peripheral autonomic blockade.

Two experiments were conducted to ascertain the cardiovascular accompaniments of differential Pavlovian jaw movement (JM) conditioning. The first examined the blood pressure (BP) changes that accompany the tachycardiac conditioned responses (CRs) associated with JM conditioning. The BP response in all instances consisted of a depressor response that was greater to the reinforced CS+ than CS-, although the magnitude of the CR was quite small. The second experiment determined the effects of peripheral autonomic antagonists on the cardiac accelerations associated with JM conditioning. It was found that the peripheral vagal antagonist methyl scopolamine completely abolished responses to both CS+ and CS-, whereas atenolol, a beta adrenergic antagonist, augmented the response, compared to saline control injections. The JM responses were also affected by the autonomic blockades, with minimal responding occurring in the scopolamine group but slightly more JM CRs in the atenolol group, compared to saline control animals. These results suggest that the major cardiovascular response to an appetitive stimulus, which evokes JM conditioning, consists of cardiac accelerations with the BP depressor responses playing a minimal, if any, role. Moreover, these conditioned cardiac increases appear to be due solely to the release of vagal inhibition.

Characterization of the neuronal changes in the medial prefrontal cortex during jaw movement and eyeblink Pavlovian conditioning in the rabbit.

Medial prefrontal (mPFC) single-unit activity was assessed in defensive (eyeblink, EB) and appetitive (jaw movement, JM) Pavlovian conditioning in the rabbit. Concomitant heart rate (HR) changes were also assessed. In a first experiment robust JM conditioned responses (CRs) were observed to a tone-water (CS+) contingency but not to tone-alone (CS-), indicating discriminative JM conditioning. However, the CS-evoked accelerative HR response was not discriminative. Nevertheless, several single-unit discharge patterns were evoked by both tone-water and tone-alone, many of which were uniquely associated with either the CS+ or CS-. In a second experiment, Three separate stimuli, consisting of tone followed by periorbital shock (tone-shock), water (tone-water), and white noise not followed by shock or water, were presented in the same paradigm. Discrimination of conditioned JM, EB and HR changes were observed, i.e. each of these behavioral responses were uniquely associated with the relevant CS presentation. Conditioned bradycardia was evoked by tone-water during the first training session, which changed to tachycardia with further training. However, conditioned bradycardia was evoked by tone-shock throughout training. Different subpopulations of mPFC cells were activated by the tone-shock and tone-water contingencies, but a small group of cells were activated by both.

Medial prefrontal cortex and pavlovian conditioning: trace versus delay conditioning.

Pavlovian eyeblink (EB) conditioning was studied in both trace and delay paradigms in rabbits (Oryctolagus cuniculus) with either medial prefrontal cortex (mPFC) lesions or sham lesions. mPFC lesions of prelimbic cortex (Brodmann's Area 32) retarded EB conditioning in the trace but not the delay paradigm. However, this effect was significant only when the conditioned stimulus (CS) was 500 rather than 100 ms in duration. Lesions of the anterior cingulate cortex (Area 24) did not affect EB conditioning in a trace paradigm. Accompanying CS-evoked heart rate slowing was attenuated under all conditions by the mPFC lesions, although this result was not always statistically significant.

Multiple response measures during classical conditioning.

Animal research assessing multiple responses during Pavlovian conditioning has revealed a dichotomy between the central nervous system (CNS) substrates for somatomotor and visceral CRs. These findings have implications for the study of clinical/applied problems in human subjects, since differences in the acquisition functions for these response systems may suggest which CNS structures are involved in various neuropsychiatric disorders. The present paper describes methods and procedures utilized to assess the somatomotor conditioned eyeblink (EB) response and accompanying visceral changes in human subjects. Methods are described for assessing concomitant EB conditioned and unconditioned responses and the accompanying heart rate, skin conductance, and respiratory changes during Pavlovian conditioning in human subjects. It is stressed that utilization of concomitant conditioning of these different response systems may lead to inferences regarding the central nervous system structures involved in a variety of different kinds of clinical problems.

Posttraining lesions of the medial prefrontal cortex impair performance of Pavlovian eyeblink conditioning but have no effect on concomitant heart rate changes in rabbits (Oryctolagus cuniculus).

The medial prefrontal cortex (mPFC) plays a critical role in conditioned autonomic adjustments but is not involved in classically conditioned somatomotor responses unless the training conditions include reversal or trace conditioning. The studies showing these effects have all used pretraining lesions. The present study assessed the effects of posttraining lesions on eyeblink (EB) and heart rate (HR) conditioned responses (CRs) in both delay and trace conditioning paradigms in the rabbit (Oryctolagus cuniculus). Posttraining lesions lowered the percentage of EB CRs during retesting compared with pretesting levels for both delay and trace conditioning. Control lesions and pretraining lesions produced no significant effects during retesting. Posttraining lesions had no effect on the HR CR. These findings suggest that a critical mechanism in the mPFC is involved in retrieval of information during EB conditioning but that the mPFC integration of autonomic and somatomotor processes is not critical to this retrieval process.