Licorice and its potential beneficial effects in common oro-dental diseases.

Licorice, the name given to the roots and stolons of Glycyrrhiza species, has been used since ancient times as a traditional herbal remedy. Licorice contains several classes of secondary metabolites with which numerous human health benefits have been associated. Recent research suggests that licorice and its bioactive ingredients such as glycyrrhizin, glabridin, licochalcone A, licoricidin, and licorisoflavan A possess potential beneficial effects in oral diseases. This paper reviews the effects of licorice and licorice constituents on both the oral microbial pathogens and the host immune response involved in common ora-dental diseases (dental caries, periodontitis, candidiasis, and recurrent aphthous ulcers). It also summarizes results of clinical trials that investigated the potential beneficial effects of licorice and its constituents for preventing/treating oro-dental diseases.

Food restriction induces long-lasting recovery of spatial memory deficits following global ischemia in delayed matching and non-matching-to-sample radial arm maze tasks.

Food restriction has been shown to be beneficial for a number of brain processes. In the current study, we characterized the impact of food restriction on hippocampal damage 70 days following ischemia. We assessed memory and cognitive flexibility of ad libitum fed (AL) and food-restricted (FR) animals using complex delayed non-matching- and matching-to-sample tasks in the radial arm maze. Our findings demonstrate that food restriction led to significant improvement of ischemia-induced memory impairments. FR ischemic animals rapidly reached comparable performance as both AL and FR sham animals in delayed-non-matching (win-shift) and matching (win-stay) radial arm maze tasks. They also made considerably fewer microchoices in the retention trials than AL ischemic animals. In contrast, AL ischemic rats showed persistent spatial memory impairments in the same paradigms. Assessment of basal and stress-induced corticosterone (CORT) secretion revealed no significant differences in baseline levels in AL and FR rats prior to or following global ischemia. However, FR animals showed a more pronounced attenuation of CORT secretion 45 min following restraint. Both FR and AL ischemic rats had comparable cell loss within CA1 and CA3 subfields of Ammon's horn (CA1 and CA3) at 70 days following reperfusion, although a trend toward increased CA3 cell survival was observed in FR ischemic rats. The functional sparing in the FR ischemic animals in the face of equivalent hippocampal cell loss suggests that food restriction somehow enhanced the efficacy of remaining hippocampal or extrahippocampal neurons following ischemia. In the current study, this phenomenon was not associated with diet- and or ischemia-related alterations of vesicular glutamate transporter 1 expression in various hippocampal regions although lower vesicular GABA transporter immunostaining was present in the CA1 stratum oriens and the CA3 stratum radiatum in FR sham and ischemic rats.

Cerebral glucose transporters expression and spatial learning in the K-ATP Kir6.2(-/-) knockout mice.

K-ATP channels formed of the Sur and Kir subunits are widely distributed in the brain. Sur1-Kir6.2 is the most common combination of K-ATP channel subunits in the brain and Kir6.2 plays an important role in glucose metabolism through pancreatic insulin secretion or hypothalamic glucose sensing. K-ATP channels have also been reported to play a role in memory processing. Therefore, the aim of the present experiment is to assess the gene and protein expression of GLUT1, GLUT3 and GLUT4 in various brain regions of Kir6.2(-/-) K-ATP knockout mice and to test their working memory performance. GLUT4 was measured using two antibodies, one recognizing an intracellular epitope and the other, an extracellular epitope. Relative to their corresponding wild type, semi-quantitative immunohistochemistry showed that GLUT4 protein expression as measured by a GLUT4 antibody recognizing an extracellular epitope was increased in the Kir6.2(-/-) K-ATP mice. However, there was only a small increase in GLUT4 labeling using the GLUT4 antibody recognizing the intracellular epitope. These results suggest a compensatory higher GLUT4 inclusion at the cellular neuronal membrane in the cerebral cortex, hippocampus and cerebellum of the Kir6.2(-/-) K-ATP knockout mice. However, there was no change in GLUT4 gene expression assessed by TaqMan PCR except for a decrease in the cerebellum of these mice. Working memory performance of the Kir6.2(-/-) K-ATP mice was disrupted at age of 12 weeks but not at 5 weeks. The mild glucose intolerance that is observed in the Kir6.2 knockout mice is unlikely to have created the memory deficits observed. Rather, in light of the effects of K-ATP channel modulators on memory, the memory deficits in the Kir6.2(-/-) K-ATP mice are more likely due to the absence of the Kir6.2 and possible disruption of the GLUT4 activity in the brain.

Glucose transporter plasticity during memory processing.

Various types of learning, including operant conditioning, induce an increase in cellular activation concomitant with an increase in local cerebral glucose utilization (LCGU). This increase is mediated by increased cerebral blood flow or changes in brain capillary density and diameter. Because glucose transporters are ultimately responsible for glucose uptake, we examined their plastic expression in response to cellular activation. In vitro and in vivo studies have demonstrated that cerebral glucose transporter 1 (GLUT1) expression consistently parallels changes in LCGU. The present study is the first to investigate the effect of memory processing on glucose transporters expression. Changes in GLUT expression produced by training in an operant conditioning task were measured in the brain of CD1 mice. Using semi-quantitative immunohistochemistry, Western blot and real time RT-PCR the cerebral GLUT1 and GLUT3 expression was quantified immediately, 220 min and 24 h following training. Relative to sham-trained and naive controls, operant conditioning training induced an immediate increase in GLUT1 immunoreactivity level in the hippocampus CA1 pyramidal cells as well as in the sensorimotor cortex. At longer post-learning delays, GLUT1 immunoreactivity decreased in the sensorimotor cortex and putamen. Parallel to the changes in protein levels, hippocampus GLUT1 mRNA level also increased immediately following learning. No effect of learning was found on hippocampal GLUT3 protein or mRNA expression. Measures of changes in glucose transporters expression present a link between cellular activation and glucose metabolism. The learning-induced localized increases in GLUT1 protein as well as mRNA levels observed in the present study confirm the previous findings that GLUT1 expression is plastic and respond to changes in cellular metabolic demands.

Methodological and conceptual issues in the use of the elevated plus-maze as a psychological measurement instrument of animal anxiety-like behavior.

There has been some suggestion that 'risk assessment' defensive behaviors in rodents might resemble some of the behavioral/somatic symptoms of generalized anxiety in humans. Although the inclusion of some risk assessment behaviors enhanced the sensitivity of the elevated plus-maze to detect subtle changes in anxiety-like behavior, there is little evidence to support the inclusion of 15 or 20 indicator variables in an analysis. Several methodological, conceptual, complexity and interpretation problems associated with the factorial validity of recently published ethologically-derived large-scale principal components analyses of elevated plus-maze behavior are examined in this review. The utility of confirmatory factor analytic work currently being conducted in our laboratory to test structural hypotheses of anxiety-relevant elevated plus-maze behavior is then discussed with a view to address some of these issues. Finally, we propose that the growing number of measured behavioral indices in the elevated plus-maze test battery be reduced, and suggest that some of the underlying constructs thought to drive behavior in the apparatus are in need of re-evaluation.

Effect of glucose and peripheral glucose regulation on memory in the elderly.

We examined changes in cognitive performance following the intake of a glucose (50 g) or saccharin solution (50 mg) in fasted elderly male and female subjects. Glucoregulation was estimated using a recovery index that was used to categorize subjects within each sex as having poor or good recovery. Elderly males with poor recovery performed worse on the Logical Memory subtest of the Wechsler Memory Scale and on the free recall or recognition parts of a work list task. The item analysis of the Logical Memory subtest showed that male subjects with poor recovery remembered less of the last items of the paragraphs after drinking saccharin while the first items were equally remembered by both groups. Glucose improved the performance of the males with good regulation for the first seven items while the performance of males with poor regulation decreased for those items under glucose. The present study support the notion that peripheral glucoregulation can influence memory performance and that the ingestion of glucose can influence certain aspects of memory functioning.

Immunohistochemical localization and quantification of glucose transporters in the mouse brain.

A family of seven facilitative glucose transporters (Glut1-5, 7 and 8) mediates the cellular uptake of glucose. In the brain, Glut2, Glut5 and Glut8 are found at relatively low levels whereas Glut1, Glut3 and Glut4 were reported in abundance in several brain regions. Using immunofluorescence, this study investigated, compared and quantified the localization of the brain major glucose transporters, Glut1, Glut3 and Glut4, in the different cerebral areas of CD1 mice. Most of the staining of Glut1, Glut3 and Glut4 in the mouse brain coincides with observations made in rats. The results confirm the cortical neuropil distribution of Glut3, the prominence of this transporter in the mossy fiber field of the hippocampus and the Glut3 and Glut4 immunostaining of the hippocampal pyramidal cell layer. The present study also reports novel localizations of the transporters such as the presence of Glut3 in neuronal perikarya, Glut4-labeled neurons in the CA3 of the hippocampus and the subiculum. In the cerebellum, Glut3 shows subcellular localization to the base of the Purkinje cell bodies near the axon hillock. Furthermore, an important population of Golgi cells was found to be strongly immunostained for Glut4 in the granular cell layer of the cerebellum. The quantification results suggest that the relative abundance of Glut1 in the frontal and motor cortices coincides well with the high-energy demands of these brain regions. However, the Glut4-selective abundance in cerebral motor areas supports its suggested role in providing the energy needed for the control of the motor activity. The reported neuropil distribution of Glut3 seems to uphold its suggested role in synaptic energy provision and neurotransmitter synthesis. We conclude that the cellular and regional distributions of the glucose transporters in the rodent brain seem to be relevant to their corresponding functions.

Memory processing and apamin induce immediate early gene expression in mouse brain.

The present study analyses the effects of learning on the spatial pattern and the time-course of changes of immediate early gene messenger RNA's (c-fos and c-jun) in mouse brain produced by training in an appetitive bar-pressing task. Activation of c-fos and c-jun after training is strictly located in the hippocampal formation and is learning-dependent. Levels of both proto-oncogene mRNAs in the trained group were 4 to 5 times higher than in the sham-conditioned group. Injections of apamin, a bee venom neurotoxin that selectively blocks a class of Ca(2+)-activated K+ channels and improves learning and memory retention, produced as compared to untrained animals a 3- to 5-fold increase of expression of c-fos and c-jun with the same pattern as that observed in the trained animals. Post-training injection of 0.2 mg/kg apamin enhanced 1.4-fold the expression of both immediate early genes in CA1, CA3 and dentate gyrus as compared to trained saline-injected mice. All these results suggest that apamin-induced increase of immediate early gene expression might be related to the apamin-induced facilitation of learning.

Effects of adrenal demedullation on the conditioned emotional response and on the memory improving action of glucose.

Results of previous experiments have demonstrated that posttraining, noncontingent ingestion of sucrose solutions, or injection of glucose solutions improve retention of various learning tasks. In the present experiment, we tested the hypothesis that this effect is due to a glucose-stimulated release of epinephrine from the adrenal medulla by testing the effect of posttraining glucose injections on retention of a conditioned emotional response (CER) in demedullated rats. In a preliminary experiment, demedullated rats were found to be deficient at acquiring the CER, but a similar deficit in sham-operated animals suggested that this was due to the surgical procedure rather than to the absence of the adrenal medulla. When appropriate shock parameters were used, posttraining glucose improved retention of the CER in a manner parallel to the effect of this treatment in normal rats. It was concluded that the memory improving effect of posttraining glucose does not involve an effect of this substance on the adrenal medulla.

Effect of glucose, glucose regulation, and word imagery value on human memory.

Changes in memory performance were examined after intake of a glucose (50 g) or saccharin (50 mg) solution in fasted men and women. Glucoregulation was estimated by using a recovery index to categorize participants within each gender as having poor or good recovery. Memory was assessed with word-learning tasks in which the imagery-evoking value of the words was systematically manipulated to yield high- and low-imagery lists. The results showed that men and women characterized as having poor glucose regulation had significantly worse memory performance under the saccharin condition. This decrement was reversed by glucose ingestion. These effects were observed for both low- and high-imagery words. This study supports the hypothesis that poor glucoregulation is associated with poor memory performance even in young healthy participants and that the ingestion of glucose can improve their memory.

Infralimbic muscarinic M1 receptors modulate anxiety-like behaviour and spontaneous working memory in mice.

RATIONALE: Spontaneous working memory and anxiety-like behaviour can be concurrently influenced following kappa 1 opioid agonist or antagonist infusions in the infralimbic (IL) area of the ventromedial prefrontal cortex (vmPFC) in CD-1 mice. OBJECTIVE: The present study sought to evaluate whether acetylcholine (ACh) muscarinic (M) receptor drugs can similarly influence these cognitive-behavioural processes in the IL cortex. METHOD: Anxiety was evaluated in the elevated plusmaze and spontaneous working memory was evaluated in the Y-maze following scopolamine, pirenzepine or McN-A-343 infusion in the IL cortex. RESULTS: In experiment 1, the non-specific muscarinic receptor antagonist, scopolamine, was anxiogenic in trial 1 (5, 10 and 20 nmol), but did not influence behaviour in trial 2 (no-injection) in the elevated plus-maze 24 h later. In week 2, scopolamine disrupted spontaneous working memory in the Y-maze at the highest dose (20 nmol). In experiment 2, pretreatment with the M1 antagonist, pirenzepine, was anxiolytic in trial 1 (5 and 10 nmol), as well as in trial 2 (no-injection) in the elevated plus-maze 24 h later (0.25, 1.25, 2.5, 5 and 10 nmol). In week 2, pirenzepine disrupted spontaneous working memory in the Y-maze (2.5, 5 and 10 nmol). In experiment 3, pretreatment with the M1 agonist, McN-A-343, was anxiogenic in trial 1 (2.5, 5, 10 and 20 nmol), as well as in trial 2 (no-injection) in the elevated plus-maze 24 h later (2.5, 5, 10 and 20 nmol). In week 2, McN-A-343 enhanced spontaneous working memory in the Y-maze (2.5, 5, 10 and 20 nmol). CONCLUSIONS: (1) Enhanced ACh transmission in the vmPFC induces anxiety in challenging environments and enhances spontaneous working memory performance. (2) Blocking or activating postsynaptic M1 receptors in the vmPFC may truncate or exaggerate, respectively, afferent anxiety-relevant information. (3) IL pirenzepine and McN-A-343 exert long-term opposite effects on aversive learning during trial 1 in the elevated plus-maze.

Dose-dependent action of glucose on memory processes in women: effect on serial position and recall priority.

Previous research has shown that glucose can enhance memory in animals and humans. In humans, the facilitative effect of glucose is best observed with declarative memory tasks in older subjects. While the memory-enhancing action of glucose is well established, the underlying physiological mechanisms and the specific aspects of memory that are modulated by glucose in humans are not well understood. The present study sought to examine the effects of glucose on memory in young women using a memory paradigm sensitive to specific encoding and retrieval strategies. The glucose dose was adjusted for the weight of each participant in order to generate a dose response curve covering most doses used in previous studies. The results showed that 300 mg/kg glucose enhanced the primacy effect as defined by the recall of the first five items of the lists. However, none of the doses of glucose produced changes in the recall priority given to primacy items. The effect of glucose appears to be localized on the recall primacy effect, suggesting that glucose acts on precise memory operations. This improvement, however, is independent of the order in which subjects recalled the words. Cholinergic drugs have been shown to alter the recall of the primacy part of word lists and this observation is consistent with the hypothesis that glucose acts on memory through an interaction with brain cholinergic systems.

The absence of effect of glucose on memory is associated with low susceptibility to the amnestic effects of scopolamine in a strain of mice.

In this series of experiments, we examined the ability of post-training glucose injections to improve memory of the Balb/cAnNCrlBR strain of mice for a bar-pressing task. We could not replicate this effect which has been demonstrated in many other strains of mice including Balb/cbyJ, a related strain. We found that the Balb/cAnNCrlBR strain of mice is also much less sensitive to the disrupting effects produced by pre- or post-training injections of the competitive cholinergic antagonist scopolamine. This strain also shows altered glucoregulation compared to the Balb/cbyJ strain. The absence of glucose effects on memory in Balb/cAnNCrlBR mice appears to be associated with decreased sensitivity to cholinergic antagonists. These results can be contrasted with previous ones obtained in a related strain, the Balb/cbyJ, in which glucose was shown to improve memory while scopolamine could easily disrupt memory processes. Taken together, these data provide additional indirect support for the hypothesis that glucose improvement of memory is closely linked to a functional interaction with central cholinergic systems. The comparison of these two strains could be the basis for a useful animal model to investigate the relationship between age-related changes in memory and central cholinergic function.

Improvement of memory for an operant response by post-training glucose in mice.

Previous experiments have shown that a post-training glucose injection can retroactively and non-contingently improve the retention of a previously learned association. To date, the memory-improving action of glucose has only been demonstrated in rats for negatively-motivated tasks. The present experiment sought to generalize these previous results by examining the effects in mice of post-training glucose injections on the retention of an operant bar-pressing response. The results show that post-training glucose can retroactively and non-contingently improve the retention of an appetitively motivated task in mice. There was a U-shaped relationship between the dose of glucose and the effect on memory similar to the ones already observed in rats using negatively motivated training. The implications of these results for an endogenous memory modulation mechanism are discussed.

Glucose regulation and cognitive functions: relation to Alzheimer's disease and diabetes.

Glucose has been found to improve memory in animals and humans. Animal research has revealed that glucose may improve memory through a facilitation of acetylcholine (ACh) synthesis and release in the brain. This glucose-related memory improvement has prompted research in elderly humans. These studies have shown that the memory-improving action of glucose depends on each individuals' blood glucose regulation. Based on these data, researchers have evaluated the effect of glucose on memory in patients with Alzheimer's disease (AD). Results demonstrated that glucose could improve memory in a subset of patients that had abnormalities in their blood glucose regulation. Interestingly, these alterations in blood glucose regulation were believed to depend on the severity of the disease process. Another line of investigation has focused on alterations in brain glucose metabolism. Both animal models and studies with Type II diabetic elderly patients have shown that altered glucose regulation impairs learning and memory processes. It is possible that in AD patients, hyperglycemia exerts a deleterious effect by potentiating the neuronal death produced by other pathological processes taking place such as amyloid deposition. Based on these data, it appears important to find the prevalence of altered glucoregulation at various stages of AD. Secondly, it may be of interest to determine prospectively whether altered glucoregulation is linked to a faster progression of the disease. Finally, if such a relationship is observed, the next logical step would be to determine whether AD patients could benefit from treatments aimed at normalizing blood glucose regulation and improving insulin sensitivity.

The hippocampal formation--orbitomedial prefrontal cortex circuit in the attentional control of active memory.

The long held view that the hippocampal formation is not only essential, but also solely responsible for declarative memory in humans (and by analogy non-human primates) has come into question. Based on extensive reciprocal connection patterns between the hippocampal formation and the orbitoventromedial prefrontal cortex in primates and rats, a central role for the hippocampal formation in the attentional control of behavior is emerging. In this paper, evidence is reviewed showing that the hippocampal-orbitomedial prefrontal cortex circuit may be involved in attentional monitoring of the internal sensorium. This attentional monitoring system, in a sense, is the working memory of viscero-emotional processing. The hippocampal formation can thus be viewed as a discrepancy detector with respect to the relative activational status of cognitive/emotional set in the orbitomedial prefrontal cortex. Discrepancies between the current representation of the internal milieu and the "just-prior" representation held "on-line" in orbitomedial prefrontal cortex associative working memory, are signaled from the hippocampus to the prefrontal cortex prospective attentional systems to activate, process, and reconcile internal (past) with external (present) environments, and finally to effectively alter active working emotional "sets" to exert cognitive-emotional control of behavior.

Concurrent modulation of anxiety and memory.

We have previously shown that the ventromedial prefrontal cortex (vmPFC) is involved in spontaneous working memory and anxiety-related behaviour in CD-1 mice. Specifically, pretrial microinjection of the kappa(1) agonist, U-69,593, in the infralimbic (IL) area of the vmPFC produced a robust anxiolytic behavioural profile in the elevated plus-maze and enhanced spontaneous working memory in the Y-maze. In the present study we sought to determine whether these effects were specific to IL kappa receptors. We hypothesized that microinjection of the kappa antagonist, norBNI, in the IL cortex would influence anxiety and spontaneous memory in an opposite direction to the effects produced by the kappa(1) agonist. In week 1, transfer-latency reference memory and anxiety were tested in the elevated plus-maze in two separate trials with an intertrial interval of 24 h. In week 2, spontaneous working memory was tested in the Y-maze followed immediately by defensive/withdrawal anxiety in the open field for one half of the animals in each group, and the other half was tested in reverse order. Pretreatment with one injection of vehicle, 1, 5 or 10 nmol/0.5 microl norBNI in the IL cortex dose-dependently reduced transfer-latencies and produced an anxiogenic behavioural profile in the first elevated plus-maze trial. Following a 24 h delay, transfer-latency reference memory was not influenced, but a robust anxiogenic behavioural profile was observed in the second no-injection anxiety trial in the elevated plus-maze relative to control animals. In week 2, the same groups of mice were again pretreated with one injection of the same doses of norBNI in the IL cortex and tested in the open field and Y-maze. NorBNI pretreatment was anxiogenic in the defensive/withdrawal anxiety test and disrupted spontaneous working memory regardless of testing order. The present results show the influence of kappa receptor modulation on anxiety induction and spontaneous working memory. These results also support the hypothesis that immediate memory processing may modulate the induction of anxiety-related behaviours.

Ethological confirmatory factor analysis of anxiety-like behaviour in the murine elevated plus-maze.

The elevated plus-maze has been used in animal research to measure anxiety since 1985 and is currently the most widely used animal model of anxiety. Since this paradigm has been the subject of several principal components analyses, it is well qualified for confirmatory factor analysis research. The current report builds on the substantial theoretical knowledge and empirical data obtained from these structural analyses with a view to obtain further progress in the evolution of our understanding of animal anxiety in the elevated plus-maze. The purpose of the present report was two-fold: (a) to test if the a piori imposition of a 3-factor model, or a competing 2-factor elevated plus-maze model, would fit our sample (n=200 CD-1 mice) data in each of two trials within an inferential confirmatory factor analytic framework; (b) provide a well-fitting model that confers indicator variables that can most effectively and parsimoniously measure underlying constructs of elevated plus-maze behaviour. Multiple model-fitting criteria were used, and issues related to data non-normality, outliers, replicability of the model, sampling error and error of approximation in the estimation of final model fit were addressed. The final 2-factor model, with estimated error covariance between two different pairs of indicator variables, was a good fit on the trial-1 data, although it was necessary to allow unprotected stretch attends to non-significantly cross-load on factor-2. A 2-factor model also fit the trial-2 data from the present analysis, although it was necessary to allow closed arm time ratio to negatively cross-load on factor-1. These results indicate that inferential hypothesis testing and model building procedures within a confirmatory factor analysis framework produces interpretable animal anxiety indices in the elevated plus-maze. Moreover, a 2-factor, rather than a 3-factor model, parsimoniously and unambiguously explained the underlying constructs of anxiety-like mouse behaviour in the elevated plus-maze in the present study. Taken together, a reduction in the growing number of behavioural indices reported in elevated plus-maze pharmacological studies is suggested.

Memory-improving action of glucose: indirect evidence for a facilitation of hippocampal acetylcholine synthesis.

The effect of a 3 g/kg glucose injection on the velocity of the sodium-dependent high-affinity choline uptake mechanism in the hippocampus was both measured in quiet control mice and in mice immediately after training in an operant bar pressing task. Glucose did not significantly change high-affinity choline uptake in resting animals. High-affinity choline uptake in the hippocampus was increased by training in the operant bar pressing task. Glucose significantly reduced the amplitude of the increase in high-affinity choline uptake observed in the trained animals. Similarly, a 3 g/kg glucose injection also attenuated the increase in high-affinity choline uptake observed in animals injected with 1 mg/kg scopolamine. Finally, a 3 g/kg glucose injection significantly attenuated the amnesia produced by a post-training 1 mg/kg scopolamine injection in mice trained for an operant bar pressing task. These results provide additional evidence for an action of glucose on hippocampal cholinergic activity under conditions of high acetylcholine demand. This action may be mediated via an increase in acetyl coenzyme A availability, one of the precursors of acetylcholine. This facilitative effect of glucose on hippocampal acetylcholine synthesis may constitute the physiological basis for its facilitative action on memory and its attenuation of scopolamine amnesia.