Pre-Exposure, But Not Overshadowing, Inhibits Nocebo Hyperalgesia.

Nocebo hyperalgesia is a pervasive problem that significantly adds to the burden of pain. Conditioning is a key mechanism of nocebo hyperalgesia and recent evidence indicates that, once established, nocebo hyperalgesia is resistant to extinction. This means that preventive strategies are critical. We therefore tested whether two novel strategies - overshadowing (Experiment 1) and pre-exposure (Experiment 2) - could inhibit conditioned nocebo hyperalgesia. Overshadowing involves introducing additional cues during conditioning that should compete with and overshadow learning about the target nocebo cue. Pre-exposure involves pre-exposing the target nocebo cue in the absence of pain, which should diminish its ability to become associated with pain later. In both studies, healthy volunteers (N = 141) received exposure to a series of electrocutaneous pain stimuli with and without a sham electrode 'activated', which they were led to believe was a genuine hyperalgesic treatment. Nocebo conditioning was achieved by pairing sham activation with high pain prior to testing at equivalent pain intensity. In both studies, standard nocebo conditioning led to clear nocebo hyperalgesia relative to natural history controls. In Experiment 1, there was no evidence that overshadowing attenuated nocebo hyperalgesia. Importantly, however, Experiment 2 found that pre-exposure successfully attenuated nocebo hyperalgesia with post hoc analysis suggesting that this effect was dose-dependent. These findings provide novel evidence that pre-exposure, but not overshadowing, could be a cheap and effective way for mitigating the substantial harm caused by conditioned nocebo hyperalgesia in clinical settings. PERSPECTIVE: Nocebo hyperalgesia causes substantial patient burden with few preventive options available. Our study found novel evidence that pre-exposing treatment cues without pain, but not overshadowing them with other cues, has the capacity to inhibit conditioned nocebo hyperalgesia. Pre-exposure may therefore be an effective preventive strategy to combat nocebo hyperalgesia.

Metabolic and behavioural effects in offspring exposed to maternal sucrose consumption: a systematic review and meta-analysis of data from rodent models.

Consumption of sugar-sweetened beverages (SSBs) during pregnancy has been associated with childhood obesity. Research in which rodent dams have been given high-fat/high-sugar diets has consistently found metabolic alterations in their offspring. However, what remains unclear is the potential impact on the developing fetus of giving sugar in isolation at concentrations similar to SSBs to the mothers. Therefore, we conducted a systematic review and meta-analysis (Protocol No: 127115 on Prospero) to identify potential relationships between maternal sucrose consumption and metabolic outcomes in offspring of rodent (rat or mouse) models. We analysed studies that provided rodent mothers dams with access to sucrose solutions (8-20% w/v) prior to conception, during pregnancy and/or lactation and that reported offspring outcomes of body weight (BW), body composition and glycaemic control. Following a systematic search of four databases (PubMed, EMBASE, Web of Science and Scopus) performed on 15 January 2019, maternal and offspring data from 15 papers were identified for inclusion. Only rat studies were identified. Meta-analyses were performed on standardised mean differences for maternal and offspring BW and fasting glucose levels, with subgroup analyses of strain, sucrose concentration, exposure period and sex of offspring. A bias towards the inclusion of only data from male offspring was identified and this limited interpretation of potential sexually dimorphic outcomes. Maternal sucrose exposure was associated with an increased risk of obesity and poor glucose disposal in adult and aged offspring.

Metabolic and behavioural effects of prenatal exposure to non-nutritive sweeteners: A systematic review and meta-analysis of rodent models.

Little is known about possible effects of maternal non-nutritive sweetener (NNS) consumption on the metabolic health of a child. Animal models of maternal NNS consumption during pregnancy or weaning have yielded widely varying results, and there appears to be no clear consensus on the consequences for offspring body weight, glycaemic control or sweet preference choices. Moreover, heterogeneity in study design has hampered a clear focus for future research relevant to human health. In an effort to bring clarity, we have conducted a systematic review and meta-analysis (protocol no: CRD42018109509) in animal models (rat or mouse) of maternal NNS feeding (compared to water or basal diet) during pre-gestation, pregnancy or lactation. Four databases were searched from inception to 15th September 2018: PubMed, EMBASE, SCOPUS and Web of Science. We present maternal and offspring data from 24 included studies, which have been quantitatively analysed after study quality assessment, to identify relationships between maternal diet and offspring body weight (BW), feeding behaviour and glycaemic control. In 11 data sets, exposure to NNS reduced maternal BW during pregnancy, with no effect on litter outcomes. Meta-analyses on offspring BW during weaning (1123 offspring) and adulthood (646 offspring) identified small decreases in BW for both sexes. Subgroup analyses revealed reductions in BW of rat, but not mouse models. High dosage appears to be a potential factor for reduced palatability that could influence BW results; however, a lack of reported data limited our ability to confirm. Despite this, and the fact many papers were predisposed to bias, the balance of evidence suggests a maternal NNS diet during pregnancy or lactation did not increase the body weight in offspring.

Early handling reduces vulnerability of rats to activity-based anorexia.

Resistance to restricted feeding with and without wheel access was tested in rats handled (H) for 20 days since birth. Weight loss produced by 1.5-hr restricted food access was less in H than in non-handled (NH) males when tested aged 41 days. At this age combining food restriction with access to a running wheel (a procedure commonly known as activity-based anorexia, ABA) produced very rapid weight loss and no effect of handling was detected. When 75-day females were tested in the same way, under the ABA procedure H rats took longer than NH controls to reach the removal criterion. Simply restricting food access in these females produced variable weight loss, without detection of any handling effect. No differences in food intake or running were detected between H and NH rats in either males or females. In conclusion, handling seems to have a direct effect on rats' later response to either food deprivation alone or to an ABA procedure.

Activity-based anorexia: ambient temperature has been a neglected factor.

Activity-based anorexia refers to the self-starvation of rats exposed to experimental conditions that combine restricted access to food with access to an activity wheel. This paper compares previous studies of this phenomenon in relation to the ambient temperatures (AT) that were employed. On this basis, and from some more direct evidence, we argue that AT is an important, but neglected, factor in activity-based anorexia research. More attention to AT is needed in future research, since its neglect threatens the validity of conclusions drawn from those studies. Furthermore, direct examination of the effect of AT on activity-based anorexia will allow a better understanding of the mechanisms underlying this phenomenon and the possible clinical implications for the treatment of human anorexia nervosa.

The role of drinking in the suppression of food intake by recent activity.

The standard activity-based anorexia procedure provides rats with access to a running wheel while restricting their access to dry food. This can produce reduced food intake and progressive weight loss. Using this procedure, in the present study (Experiment 1) the authors found changes in drinking patterns both in the period of high activity preceding food access and during the feeding period. Varying the procedure by providing wet mash (Experiment 2) or by prior adaptation to a drinking schedule (Experiment 3) prevented the self-starvation effect. These results indicate the importance of drinking when analyzing the effect of recent activity on food intake.

Resistance to extinction of conditioned odor perceptions: evaluative conditioning is not unique.

A tasteless odor will smell sweeter after being sampled by mouth with sucrose and will smell sourer after being sampled with citric acid. This tasty-smell effect was found in experiments that compared odor-taste and color-taste pairings. Using odors and colors with minimal taste (Experiment 1), the authors found that repeated experience of odor-taste mixtures produced conditioned changes in odor qualities that were unaffected by intermixed color-taste trials (Experiment 2). An extinction procedure, consisting of postconditioning presentations of the odor in water, had no detectable effect on the changed perception of an odor (Experiments 3 and 4). In contrast, this procedure altered judgments about the expected taste of colored solutions. Evaluative conditioning (conditioned changes in liking) is claimed to be resistant to extinction. However, these results suggest that resistance to extinction in odors is related to the way they are encoded rather than to their hedonic properties.

Confusing tastes and smells: how odours can influence the perception of sweet and sour tastes.

This study investigated the relationship between perception of an odour when smelled and the taste of a solution to which the odour is added as a flavorant. In Experiment 1 (E1) sweetness, sourness, liking and intensity ratings were obtained for 20 odours. Taste ratings were then obtained for sucrose solutions to which the odours had been added as flavorants. Certain odours were found to enhance tasted sweetness while others suppressed it. The degree to which an odour smelled sweet was the best predictor of the taste ratings. These findings were extended in Experiment 2 (E2), which included a second tastant, citric acid, and employed four odours from E1. The most sweet smelling odour, caramel, was found to suppress the sourness of citric acid and, as in E1, to enhance the sweetness of sucrose. Again, odours with low sweetness suppressed the sweetness of tasted sucrose. The study demonstrated that the effects of odours on taste perception are not limited to sweetness enhancement and apply to sour as well as sweet tastes. The overall pattern of results is consistent with an explanation of the taste properties of odours in terms of prior flavour-taste associations.

Sex differences in the relationship between activity and weight loss in the rat.

Access to a running wheel combined with restricted feeding produced body weight loss at an equivalent rate in male and female litter-mate rats (Experiment 1). Thus, despite weighing less and running more, females were not more vulnerable to this procedure. When factors influencing weight loss were varied, no sex difference was found in adaptation to a new feeding schedule or in the effect of single versus group housing (Experiment 2). The apparent critical difference was that body weight loss increased running in males but not in females (Experiment 3). In all rats, rapid recovery of body weight occurred when food access was no longer restricted (Experiment 1), suggesting that "activity-based anorexia" is a misnomer for weight loss by rats in a running wheel.

Weight loss in rats produced by running: effects of prior experience and individual housing.

To investigate factors affecting activity-based anorexia (ABA) or activity-stress (AS), rats were given 2-hr access to a running wheel immediately prior to their daily 1.5-hr food access during the light cycle. This produced a reduction in food intake, a steady increase in running, and a large drop in body weight with a prolonged delay before weight recovery began. Experiment 1 found that these effects were reduced in rats with prior experience of eating at this time of day. In contrast, prior experience of running in the wheel when on ad lib food enhanced these effects in Experiment 2, where a subsequent change for half the subjects to individual housing produced a further decrease in body weight. The latter factor was investigated from the outset of Experiment 3 and again individually housed rats showed greater weight loss than did group-housed rats. This experiment also found that in rats of the same age a low initial body weight predicts greater vulnerability to ABA. It was concluded that ABA results from activity-induced reduction of feeding, which prolongs adaptation to a new feeding schedule and is accentuated by social isolation.

Activity-based anorexia in rats as failure to adapt to a feeding schedule.

Progressive weight loss resulting from restriction to a daily 1.5-hr feeding period and access to a running wheel, the Activity-Based Anorexia (ABA) effect, was obtained in 3 experiments. However, bodyweight recovered when adaptation to the feeding schedule preceded access to the wheel (Experiment 1), when feeding was at the start of the dark period (Experiments 2 and 3), and when wheel access was denied in the 4 hr before food (Experiment 4). It was concluded that ABA results from interference with adaptation to a new feeding schedule due to the development of anticipatory behavior.

Context dependency of conditioned aversions to water and sweet tastes.

Three experiments exposed rats (Rattus norvegicus) to a discriminative conditioning procedure whereby a specific fluid was followed by lithium in one environment but not in another. This produced context-specific aversion to water, as detected by 2-bottle tests in Experiment 1, and a context-dependent saccharin aversion, which was unaffected by context extinction, in Experiment 2. Experiment 3 found that sucrose preexposure increased contextual control over the aversion established by sucrose-lithium pairings but had no effect on the target context. By contrast, target context exposure during conditioning reduced aversion to this context but did not affect contextual control of the sucrose aversion. In conclusion, depending on the conditioning procedures, contextual control of a taste aversion can be independent of the context's Pavlovian properties.

Taste-potentiated odor aversion learning based on amphetamine.

This study sought to determine whether a taste can potentiate a conditioned odor aversion based on amphetamine as well as those based on lithium. A taste-potentiated odor aversion (TPOA) based on lithium was obtained in Experiment 1 only with a low concentration of an almond odor. This concentration was used in Experiment 2 where the taste, 0.1% saccharin, potentiated an odor aversion based on 1 mg/kg d-amphetamine. This was replicated in Experiment 3 where potentiation was found with doses of both 1 and 3 mg/kg amphetamine, and no effect of dose was detected. It was concluded that TPOA learning is not restricted to drugs such as lithium that produce conditioned unpalatability as well as conditioned aversions to a taste, because amphetamine does not produce conditioned unpalatability at the doses used here. Furthermore, because in Experiment 3 postconditioning extinction of the saccharin aversion removed the potentiation effect, it appears that this form of TPOA may depend on an association between the odor and taste, as proposed by within-compound theory.

Prevalence of anticipatory nausea and other side-effects in cancer patients receiving chemotherapy.

98 patients receiving chemotherapy for cancer were interviewed to determine the prevalence of anticipatory nausea and vomiting, anxiety and dietary changes. Among those who had received at least four treatments 41% reported at least mild anticipatory nausea (AN). For 24% this was a moderate to severe problem, which was significantly associated with a high level of anxiety about treatment. Prevalence at this level was independent of whether the subject was receiving treatment as an in- or an outpatient. Anticipatory vomiting (AV) was reported by only 12 patients, of whom 11 were women; this was the only effect of gender found in the sample. Independence between moderate AN and AV was also suggested by a difference in type of event triggering the effect: predominantly odours for AN and thoughts of the treatment for AV. Changes in diet after commencing chemotherapy were reported by 50% of patients who had received at least four treatments. These most commonly took the form of aversions to meat and then to coffee, and were attributed most frequently to changes in taste and then to loss of appetite.

Potentiation by a taste of toxicosis-based context conditioning: effects of varying the test fluid.

Rats that drank water in a distinctive environment and were then injected with lithium chloride (water-lithium condition) were compared with those given an added taste on those conditioning sessions (sucrose-lithium condition). In three experiments this taste potentiated a conditioned aversion to the context, as measured by suppression of intake of another solution: either a novel sour taste (Experiments 1 and 2) or a familiar saline solution (Experiment 3). In contrast, this potentiation effect was not detected when subjects were tested with water, whether a high or low dose of lithium was used (Experiment 2). Instead, in Experiments 1 and 2 water-lithium subjects drank less water than did the sucrose-lithium subjects on such tests i.e. an apparent overshadowing effect, which was the opposite outcome to that found previously using almost identical procedures. Intake on recovery sessions in another context suggested that, when water is used as the test fluid, potentiation can be masked by two factors: a context-dependent aversion to water in water-lithium subjects, and a conditioned inhibition effect of water in sucrose-lithium subjects. These may account for previous failures to detect potentiation of context conditioning.

Early experience and reinforcer quality in delayed flavour-food learning in the rat.

The ability of hungry rats to associate flavours with the consequences of ingesting glucose solutions was studied in three experiments. Experiment 1 used a procedure in which on some days one flavour, e.g. cinnamon, was presented and followed after 20 min by 20% glucose, while on other days a second flavour, e.g. wintergreen, was presented, but not followed by any event. During this training, subjects who received quinine-tainted glucose increased their consumption of the predictive flavour relative to groups given no quinine, but quinine tainting did not affect conditioned preference for the predictive flavour in choice tests. With the aim of discovering whether prior experience of a variety of foods improves ability to learn new flavour-calorie associations, Experiment 2 and used a similar procedure to compare subjects raised on a varied diet ("supermarket" rats) with controls previously given only chow. Contrary to expectation, the supermarket rats showed some impairment both on this delay task and, in Experiment 3, on one using a "mixtures" procedure. This involved presenting a mixture of one cue flavour with glucose-quinine on some days and a mixture of a second cue with an equally palatable saccharin solution on other days. Acquisition was particularly rapid in control subjects, reaching asymptote after only two flavour-glucose pairings. It was concluded that neither a decrease in palatability, as in Experiment 1, nor prior experience with a range of foods, as in Experiments 2 and 3, improve a rat's ability to associate a new flavour with calories.

Effects of basolateral amygdala lesions on taste aversions produced by lactose and lithium chloride in the rat.

In Experiment 1, intact rats were given either lactose or sucrose solutions. Although on first exposure they readily consumed lactose, its ingestion produced a conditioned taste avoidance which was partly extinguished by repeated sucrose exposure after lactose conditioning. In Experiment 2, rats with large bilateral electrolytic lesions of the basolateral amygdala and those with either sham or no operations were given two pairings of saline with LiCl injections (upper gastrointestinal tract discomfort) and in a separate condition access to high levels of lactose (lower gastrointestinal tract discomfort). Conditioned taste avoidances were measured both by two-bottle tests and by video recordings of the rats' orofacial and somatic responses. The lesions attenuated LiCl-induced taste aversion but not lactose-induced taste avoidance, results demonstrating that taste avoidance can occur without the basolateral amygdala. The results suggested that aversions based on distaste can be distinguished from avoidances based on danger, not only in terms of orofacial responses but also in terms of their neuroanatomical substrate.

Can rats learn to associate a flavour with the delayed delivery of food?

Associations between a specific flavour and access to food were studied using a discrimination procedure devised by Holman (1975). This involved giving rats one flavour (e.g. cinnamon) of saccharin solution on some days, and following this by delivery of food, and a second flavour (e.g. wintergreen) on other days which was never followed by food. Experiment 1 used glucose delivered after a 30-min delay and a slight increase in preference for the paired flavour was detected. Using a 20-min delay Experiment 2 varied the kind of food used; some evidence for discrimination learning was again found in the glucose group, but there was no evidence that rats could associate a flavour with starch solution or solid chow over this delay. To check that the general procedure was a sensitive one, in Experiment 3 one flavour was added to glucose i.e. without delay, and this produced large shifts in a subsequent preference test. Overall the results threw doubt on claims that rats as readily form flavour-calorie associations over delays as they do flavour-toxicosis associations.