Adolescent food insecurity in female rodents and susceptibility to diet-induced obesity.

Food insecurity is defined as having limited or uncertain access to nutritious foods, and adolescent food insecurity is associated with obesity and disordered eating behaviors in humans. We developed a rodent model of adolescent food insecurity to determine whether adolescent food insecurity per se promotes increased susceptibility to diet-induced obesity and altered eating behaviors during adulthood. Female juvenile Wistar rats were singly housed and assigned to three experimental diets: food-secure with standard chow (CHOW), food-secure with a high-fat/sugar Western diet (WD), and food-insecure with WD (WD-FI). Food-secure rats (CHOW and WD) received meals at fixed feeding times (9:00, 13:00, and 16:00). WD-FI rats received meals at unpredictable intervals of the above-mentioned feeding times but had isocaloric amounts of food to WD. We investigated the impact of adolescent food insecurity on motivation for sucrose (Progressive Ratio), approach-avoidance behavior for palatable high-fat food (Approach-Avoidance task), and susceptibility to weight gain and hyperphagia when given an obesogenic choice diet. Secondary outcomes were the effects of food insecurity during development on anxiety-like behaviors (Open Field and Elevated Plus Maze) and learning and memory function (Novel Location Recognition task). Rodents with adolescent food insecurity showed a greater trend of weight gain and significantly increased fat mass and liver fat accumulation on an obesogenic diet in adulthood, despite no increases in motivation for sucrose or high-fat food. These data suggest that adolescent unpredictable food access increases susceptibility to diet-induced fat gain without impacting food motivation or food intake in female rodents. These findings are among a small group of recent studies modeling food insecurity in rodents and suggest that adolescent food insecurity in females may have long-term implications for metabolic physiology later in life.

AgRP neuron activity promotes associations between sensory and nutritive signals to guide flavor preference.

OBJECTIVE: The learned associations between sensory cues (e.g., taste, smell) and nutritive value (e.g., calories, post-ingestive signaling) of foods powerfully influences our eating behavior [1], but the neural circuits that mediate these associations are not well understood. Here, we examined the role of agouti-related protein (AgRP)-expressing neurons - neurons which are critical drivers of feeding behavior [2; 3] - in mediating flavor-nutrient learning (FNL). METHODS: Because mice prefer flavors associated with AgRP neuron activity suppression [4], we examined how optogenetic stimulation of AgRP neurons during intake influences FNL, and used fiber photometry to determine how endogenous AgRP neuron activity tracks associations between flavors and nutrients. RESULTS: We unexpectedly found that tonic activity in AgRP neurons during FNL potentiated, rather than prevented, the development of flavor preferences. There were notable sex differences in the mechanisms for this potentiation. Specifically, in male mice, AgRP neuron activity increased flavor consumption during FNL training, thereby strengthening the association between flavors and nutrients. In female mice, AgRP neuron activity enhanced flavor-nutrient preferences independently of consumption during training, suggesting that AgRP neuron activity enhances the reward value of the nutrient-paired flavor. Finally, in vivo neural activity analyses demonstrated that acute AgRP neuron dynamics track the association between flavors and nutrients in both sexes. CONCLUSIONS: Overall, these data (1) demonstrate that AgRP neuron activity enhances associations between flavors and nutrients in a sex-dependent manner and (2) reveal that AgRP neurons track and rapidly update these associations. Taken together, our findings provide new insight into the role of AgRP neurons in assimilating sensory and nutritive signals for food reinforcement.

AgRP neuron activity promotes associations between sensory and nutritive signals to guide flavor preference.

Objective: The learned associations between sensory cues (e.g., taste, smell) and nutritive value (e.g., calories, post-ingestive signaling) of foods powerfully influences our eating behavior [1], but the neural circuits that mediate these associations are not well understood. Here, we examined the role of agouti-related protein (AgRP)-expressing neurons - neurons which are critical drivers of feeding behavior [2; 3] - in mediating flavor-nutrient learning (FNL). Methods: Because mice prefer flavors associated with AgRP neuron activity suppression [4], we examined how optogenetic stimulation of AgRP neurons during intake influences FNL, and used fiber photometry to determine how endogenous AgRP neuron activity tracks associations between flavors and nutrients. Results: We unexpectedly found that tonic activity in AgRP neurons during FNL potentiated, rather than prevented, the development of flavor preferences. There were notable sex differences in the mechanisms for this potentiation. Specifically, in male mice, AgRP neuron activity increased flavor consumption during FNL training, thereby strengthening the association between flavors and nutrients. In female mice, AgRP neuron activity enhanced flavor-nutrient preferences independently of consumption during training, suggesting that AgRP neuron activity enhances the reward value of the nutrient-paired flavor. Finally, in vivo neural activity analyses demonstrated that acute AgRP neuron dynamics track the association between flavors and nutrients in both sexes. Conclusions: Overall, these data (1) demonstrate that AgRP neuron activity enhances associations between flavors and nutrients in a sex-dependent manner and (2) reveal that AgRP neurons track and update these associations on fast timescales. Taken together, our findings provide new insight into the role of AgRP neurons in assimilating sensory and nutritive signals for food reinforcement.

Ventral hippocampus neurons encode meal-related memory.

The ability to encode and retrieve meal-related information is critical to efficiently guide energy acquisition and consumption, yet the underlying neural processes remain elusive. Here we reveal that ventral hippocampus (HPCv) neuronal activity dynamically elevates during meal consumption and this response is highly predictive of subsequent performance in a foraging-related spatial memory task. Targeted recombination-mediated ablation of HPCv meal-responsive neurons impairs foraging-related spatial memory without influencing food motivation, anxiety-like behavior, or escape-mediated spatial memory. These HPCv meal-responsive neurons project to the lateral hypothalamic area (LHA) and single-nucleus RNA sequencing and in situ hybridization analyses indicate they are enriched in serotonin 2a receptors (5HT2aR). Either chemogenetic silencing of HPCv-to-LHA projections or intra-HPCv 5HT2aR antagonist yielded foraging-related spatial memory deficits, as well as alterations in caloric intake and the temporal sequence of spontaneous meal consumption. Collective results identify a population of HPCv neurons that dynamically respond to eating to encode meal-related memories.

Chronic experience with unpredictable food availability promotes food reward, overeating, and weight gain in a novel animal model of food insecurity.

Ubiquitous, easy access to food is thought to promote obesity in the modern environment. However, people coping with food insecurity have limited, unpredictable food access and are also prone to obesity. Causal factors linking food insecurity and obesity are not understood. In this study we describe an animal model to investigate biopsychological impacts of the chronic unpredictability inherent in food insecurity. Female rats were maintained on a 'secure' schedule of highly predictable 4x/day feedings of uniform size, or an 'insecure' schedule delivering the same total food over time but frequently unpredictable regarding how much, if any, food would arrive at each scheduled feeding. Subgroups of secure and insecure rats were fed ordinary chow or high-fat/high-sugar (HFHS) chow to identify separate and combined effects of insecurity and diet quality. Insecure chow-fed rats, relative to secure chow-fed rats, were hyperactive and consumed more when provided a palatable liquid diet. Insecure HFHS-fed rats additionally had higher progressive ratio breakpoints for sucrose, increased meal size, and subsequently gained more weight during 8 days of ad libitum HFHS access. Insecurity appeared to maintain a heightened attraction to palatable food that habituated in rats with secure HFHS access. In a second experiment, rats fed ordinary chow on the insecure schedule subsequently gained more weight when provided ad libitum chow, showing that prior insecurity per se promoted short-term weight gain in the absence of HFHS food. We propose this to be a potentially useful animal model for mechanistic research on biopsychological impacts of insecurity, demonstrating that chronic food uncertainty is a factor promoting obesity.

The Role of Post-Ingestive Feedback in the Development of an Enhanced Appetite for the Orosensory Properties of Glucose over Fructose in Rats.

The simple sugars glucose and fructose share a common "sweet" taste quality mediated by the T1R2+T1R3 taste receptor. However, when given the opportunity to consume each sugar, rats learn to affectively discriminate between glucose and fructose on the basis of cephalic chemosensory cues. It has been proposed that glucose has a unique sensory property that becomes more hedonically positive through learning about the relatively more rewarding post-ingestive effects that are associated with glucose as compared to fructose. We tested this theory using intragastric (IG) infusions to manipulate the post-ingestive consequences of glucose and fructose consumption. Food-deprived rats with IG catheters repeatedly consumed multiple concentrations of glucose and fructose in separate sessions. For rats in the "Matched" group, each sugar was accompanied by IG infusion of the same sugar. For the "Mismatched" group, glucose consumption was accompanied by IG fructose, and vice versa. This condition gave rats orosensory experience with each sugar but precluded the differential post-ingestive consequences. Following training, avidity for each sugar was assessed in brief access and licking microstructure tests. The Matched group displayed more positive evaluation of glucose relative to fructose than the Mismatched group. A second experiment used a different concentration range and compared responses of the Matched and Mismatched groups to a control group kept naïve to the orosensory properties of sugar. Consistent with results from the first experiment, the Matched group, but not the Mismatched or Control group, displayed elevated licking responses to glucose. These experiments yield additional evidence that glucose and fructose have discriminable sensory properties and directly demonstrate that their different post-ingestive effects are responsible for the experience-dependent changes in the motivation for glucose versus fructose.

No evidence that portion size influences food consumption in male Sprague Dawley rats.

In studies of eating behavior that have been conducted in humans, the tendency to consume more when given larger portions of food, known as the portion size effect (PSE), is one of the most robust and widely replicated findings. Despite this, the mechanisms that underpin it are still unknown. In particular, it is unclear whether the PSE arises from higher-order social and cognitive processes that are unique to humans or, instead, reflects more fundamental processes that drive feeding, such as conditioned food-seeking. Importantly, studies in rodents and other animals have yet to show convincing evidence of a PSE. In this series of studies, we used several methods to test for a PSE in adult male Sprague Dawley rats. Our approaches included using visually identifiable portions of a palatable food; training on a plate cleaning procedure; providing portion sizes of food pellets that were signaled by auditory and visual food-predictive cues; providing food with amorphous shape properties; and providing standard chow diet portions in home cages. In none of these manipulations did larger portions increase food intake. In summary, our data provide no evidence that a PSE is present in male Sprague Dawley rats, and if it is, it is more nuanced, dependent on experimental procedure, and/or smaller in size than it is in humans. In turn, these findings suggest that the widely-replicated PSE in humans may be more likely to reflect higher-order cognitive and social processes than fundamental conditioned behaviors.

Portion size influences intake in Samburu Kenyan people not exposed to the Western obesogenic environment.

For people in the modernized food environment, external factors like food variety, palatability, and ubiquitous learned cues for food availability can overcome internal, homeostatic signals to promote excess intake. Portion size is one such external cue; people typically consume more when served more, often without awareness. Though susceptibility to external cues may be attributed to the modernized, cue-saturated environment, there is little research on people living outside that context, or with distinctly different food norms. We studied a sample of Samburu people in rural Kenya who maintain a traditional, semi-nomadic pastoralist lifestyle, eat a very limited diet, and face chronic food insecurity. Participants (12 male, 12 female, aged 20-74, mean BMI = 18.4) attended the study on two days and were provided in counterbalanced order an individual serving bowl containing 1.4 or 2.3 kg of a familiar bean and maize stew. Amount consumed was recorded along with post-meal questions in their dialect about their awareness of intake amount. Data were omitted from two participants who consumed the entire portion in a session. Even though the 'smaller' serving was a very large meal, participants consumed 40% more when given the larger serving, despite being unable to reliably identify which day they consumed more food. This result in the Samburu demonstrates the portion size effect is not a by-product of the modern food environment and may represent a more fundamental feature of human dietary psychology.

The convergence of psychology and neurobiology in flavor-nutrient learning.

Flavor evaluation is influenced by learning from experience with foods. One main influence is flavor-nutrient learning (FNL), a Pavlovian process whereby a flavor acts as a conditioned stimulus (CS) that becomes associated with the postingestive effects of ingested nutrients (the US). As a result that flavor becomes preferred and intake typically increases. This learning powerfully influences food choice and meal patterning. This paper summarizes how research elucidating the physiological and neural substrates of FNL has progressed in parallel with work characterizing how FNL affects perception, motivation, and behavior. The picture that emerges from this work is of a robust system of appetition (a term coined by Sclafani in contrast to the better-understood satiation signals) whereby ingested nutrients sensed in the gut evoke positive motivational responses. Appetition signals act within a meal to promote continued intake in immediate response to gut feedback, and act in the longer term to steer preference towards sensory cues that predict nutritional consequences.

Initial subjective reward to alcohol in Sprague-Dawley rats.

Initial subjective response to the rewarding properties of alcohol predicts voluntary consumption and the risk for alcohol use disorders. We assessed the initial subjective reward to alcohol in rats using a single exposure conditioned place preference (SE-CPP) paradigm. Sprague-Dawley rats demonstrate preference for a context paired with a single systemic injection of ethanol (1.0 g/kg, delivered intraperitoneally). However, expression of SE-CPP in males depended on pairing ethanol with the first exposure of two (ethanol; saline) to the conditioning apparatus and procedures, while conditioning day did not appreciably affect SE-CPP in females, consistent with the view that females experience heightened addiction vulnerability. This model offers researchers a high throughput assay for investigating factors that influence alcohol reward and may point the way toward more effective prevention and treatment efforts.

Sensory-specific satiety is intact in rats made obese on a high-fat high-sugar choice diet.

Sensory-specific satiety (SSS) is the temporary decreased pleasantness of a recently eaten food, which inhibits further eating. Evidence is currently mixed whether SSS is weaker in obese people, and whether such difference precedes or follows from the obese state. Animal models allow testing whether diet-induced obesity causes SSS impairment. Female rats (n = 24) were randomly assigned to an obesogenic high-fat, high-sugar choice diet or chow-only control. Tests of SSS involved pre-feeding a single palatable, distinctively-flavored food (cheese- or cocoa-flavored) prior to free choice between both foods. Rats were tested for short-term SSS (2 h pre-feeding immediately followed by 2 h choice) and long-term SSS (3 day pre-feeding prior to choice on day 4). In both short- and long-term tests rats exhibited SSS by shifting preference towards the food not recently eaten. SSS was not impaired in obese rats. On the contrary, in the long-term tests they showed stronger SSS than controls. This demonstrates that neither the obese state nor a history of excess energy consumption fundamentally causes impaired SSS in rats. The putative impaired SSS in obese people may instead reflect a specific predisposition, properties of the obesogenic diet, or history of restrictive dieting and bingeing.

Clinical Pharmacist Management of Bacteremia in a Community Hospital Emergency Department.

BACKGROUND: Bacteremia is a serious condition that leads to high morbidity and mortality. Data describing pharmacist involvement in the management of bacteremia in the emergency department are lacking. OBJECTIVE: To determine if pharmacist involvement in the management of bacteremia in the emergency department (ED) led to an increase in appropriate treatment of bacteremia as well as improvements in patient outcomes. METHODS: The primary outcome of this retrospective cohort study was the rate of appropriate treatment of bacteremia. Secondary outcomes included the rate of unplanned, infectious disease-related 90-day admission or readmission to the ED or hospital as well as infectious disease-related 90-day mortality. All patients seen in the ED and subsequently discharged who had a positive blood culture determined not to be a contaminant were included in the study. Patients were analyzed in 2 cohorts: those that were physician managed (107 patients) and those that were pharmacist managed (138 patients). RESULTS: In the physician-managed cohort, 50 of 107 (47%) patients were treated appropriately compared with 131 of 138 (95%) patients in the pharmacist-managed cohort ( P < 0.0001). There was also a decrease in attributable 90-day admission or readmission in pharmacist-managed patients, which occurred in 4 of 138 patients (2.9%) versus the physician-managed patient cohort in which 13 of 107 patients (12.1%) were readmitted ( P = 0.01). There was no difference in mortality between the groups ( P = 0.8337). CONCLUSION: Pharmacist involvement in the management of bacteremia in the ED was associated with higher rates of appropriate treatment and a corresponding decrease in the rates of attributable 90-day admission or readmission to the hospital or ED.

Modern 'junk food' and minimally-processed 'natural food' cafeteria diets alter the response to sweet taste but do not impair flavor-nutrient learning in rats.

Animals learn to prefer and increase consumption of flavors paired with postingestive nutrient sensing. Analogous effects have been difficult to observe in human studies. One possibility is experience with the modern, processed diet impairs learning. Food processing manipulates flavor, texture, sweetness, and nutrition, obscuring ordinary correspondences between sensory cues and postingestive consequences. Over time, a diet of these processed 'junk' foods may impair flavor-nutrient learning. This 'flavor-confusion' hypothesis was tested by providing rats long-term exposure to cafeteria diets of unusual breadth (2 or 3 foods per day, 96 different foods over 3 months, plus ad libitum chow). One group was fed processed foods (PF) with added sugars/fats and manipulated flavors, to mimic the sensory-nutrient properties of the modern processed diet. Another group was fed only 'natural' foods (NF) meaning minimally-processed foods without manipulated flavors or added sugars/fats (e.g., fresh fruits, vegetables, whole grains) ostensibly preserving the ordinary correspondence between flavors and nutrition. A CON group was fed chow only. In subsequent tests of flavor-nutrient learning, PF and NF rats consistently acquired strong preferences for novel nutrient-paired flavors and PF rats exhibited enhanced learned acceptance, contradicting the 'flavor-confusion' hypothesis. An unexpected finding was PF and NF diets both caused lasting reduction in ad lib sweet solution intake. Groups did not differ in reinforcing value of sugar in a progressive ratio task. In lick microstructure analysis the NF group paradoxically showed increased sucrose palatability relative to PF and CON, suggesting the diets have different effects on sweet taste evaluation.

Enhanced flavor-nutrient conditioning in obese rats on a high-fat, high-carbohydrate choice diet.

Through flavor-nutrient conditioning rats learn to prefer and increase their intake of flavors paired with rewarding, postingestive nutritional consequences. Since obesity is linked to altered experience of food reward and to perturbations of nutrient sensing, we investigated flavor-nutrient learning in rats made obese using a high fat/high carbohydrate (HFHC) choice model of diet-induced obesity (ad libitum lard and maltodextrin solution plus standard rodent chow). Forty rats were maintained on HFHC to induce substantial weight gain, and 20 were maintained on chow only (CON). Among HFHC rats, individual differences in propensity to weight gain were studied by comparing those with the highest proportional weight gain (obesity prone, OP) to those with the lowest (obesity resistant, OR). Sensitivity to postingestive food reward was tested in a flavor-nutrient conditioning protocol. To measure initial, within-meal stimulation of flavor acceptance by post-oral nutrient sensing, first, in sessions 1-3, baseline licking was measured while rats consumed grape- or cherry-flavored saccharin accompanied by intragastric (IG) water infusion. Then, in the next three test sessions they received the opposite flavor paired with 5 ml of IG 12% glucose. Finally, after additional sessions alternating between the two flavor-infusion contingencies, preference was measured in a two-bottle choice between the flavors without IG infusions. HFHC-OP rats showed stronger initial enhancement of intake in the first glucose infusion sessions than CON or HFHC-OR rats. OP rats also most strongly preferred the glucose-paired flavor in the two-bottle choice. These differences between OP versus OR and CON rats suggest that obesity is linked to responsiveness to postoral nutrient reward, consistent with the view that flavor-nutrient learning perpetuates overeating in obesity.

In search of flavour-nutrient learning. A study of the Samburu pastoralists of North-Central Kenya.

Much of our dietary behaviour is learned. In particular, one suggestion is that 'flavour-nutrient learning' (F-NL) influences both choice and intake of food. F-NL occurs when an association forms between the orosensory properties of a food and its postingestive effects. Unfortunately, this process has been difficult to evaluate because F-NL is rarely observed in controlled studies of adult humans. One possibility is that we are disposed to F-NL. However, learning is compromised by exposure to a complex Western diet that includes a wide range of energy-dense foods. To test this idea we explored evidence for F-NL in a sample of semi-nomadic pastoralists who eat a very limited diet, and who are lean and food stressed. Our Samburu participants (N = 68) consumed a sensory-matched portion (400 g) of either a novel low (0.72 kcal/g) or higher (1.57 kcal/g) energy-dense semi-solid food on two training days, and an intermediate version on day 3. Before and after each meal we measured appetite and assessed expected satiation and liking for the test food. We found no evidence of F-NL. Nevertheless, self-reported measures were very consistent and, as anticipated, expected satiation increased as the test food became familiar (expected-satiation drift). Surprisingly, we observed insensitivity to the effects of test-meal energy density on measures of post-meal appetite. To explore this further we repeated a single training day using participants (N = 52) from the UK. Unlike in the Samburu, the higher energy-dense meal caused greater suppression of appetite. These observations expose interesting cross-cultural differences in sensitivity to the energy content of food. More generally, our work illustrates how measures can be translated to assess different populations, highlighting the potential for further comparisons of this kind.

Sensory-specific appetition: Postingestive detection of glucose rapidly promotes continued consumption of a recently encountered flavor.

It is generally thought that macronutrients stimulate intake when sensed in the mouth (e.g., sweet taste) but as food enters the GI tract its effects become inhibitory, triggering satiation processes leading to meal termination. Here we report experiments extending recent work (see Zukerman et al., 2011 [1]) showing that under some circumstances nutrients sensed in the gut produce a positive feedback effect, immediately promoting continued intake. In one experiment, rats with intragastric (IG) catheters were accustomed to consuming novel flavors in saccharin daily while receiving water infused IG (5ml/15min). The very first time glucose (16% w/w) was infused IG instead of water, intake accelerated within 6min of infusion onset and total intake increased 29% over baseline. Experiment 2 replicated this stimulatory effect with glucose infusion but not fructose nor maltodextrin. Experiment 3 showed that the immediate intake stimulation is specific to the flavor accompanying the glucose infusion. Rats were accustomed to flavored saccharin being removed and replaced with the same or a different flavor. When glucose infusion accompanied the first bottle, intake from the second bottle was stimulated only when it contained the same flavor, not when the flavor switched. Thus we confirm not only that glucose sensed postingestively can have a rapid, positive feedback effect ('appetition' as opposed to 'satiation') but that it is sensory-specific, promoting continued intake of a recently encountered flavor. This sensory-specific motivation may represent an additional psychobiological influence on meal size, and further, has implications for the mechanisms of learned flavor-nutrient associations.

Rats acquire stronger preference for flavors consumed towards the end of a high-fat meal.

Rats learn to prefer flavors associated with postingestive effects of nutrients. The physiological signals underlying this postingestive reward are unknown. We have previously shown that rats readily learn to prefer a flavor that was consumed early in a multi-flavored meal when glucose is infused intragastrically (IG), suggesting rapid postingestive reward onset. The present experiments investigate the timing of postingestive fat reward, by providing distinctive flavors in the first and second halves of meals accompanied by IG fat infusion. Learning stronger preference for the earlier or later flavor would indicate when the rewarding postingestive effects are sensed. Rats consumed sweetened, calorically-dilute flavored solutions accompanied by IG high-fat infusion (+ sessions) or water (- sessions). Each session included an "Early" flavor for 8min followed by a "Late" flavor for 8min. Learned preferences were then assessed in two-bottle tests (no IG infusion) between Early(+) vs. Early(-), Late(+) vs. Late(-), Early(+) vs. Late(+), and Early(-) vs. Late(-). Rats only preferred Late(+), not Early(+), relative to their respective (-) flavors. In a second experiment rats trained with a higher fat concentration learned to prefer Early(+) but more strongly preferred Late(+). Learned preferences were evident when rats were tested deprived or recently satiated. Unlike with glucose, ingested fat appears to produce a slower-onset rewarding signal, detected later in a meal or after its termination, becoming more strongly associated with flavors towards the end of the meal. This potentially contributes to enhanced liking for dessert foods, which persists even when satiated.

Investigation into the specificity of angiotensin II-induced behavioral desensitization.

Angiotensin II (AngII) plays a key role in maintaining body fluid homeostasis. The physiological and behavioral effects of central AngII include increased blood pressure and fluid intake. In vitro experiments demonstrate that repeated exposure to AngII reduces the efficacy of subsequent AngII, and behavioral studies indicate that prior icv AngII administration reduces the dipsogenic response to AngII administered later. Specifically, rats given a treatment regimen of three icv injections of a large dose of AngII, each separated by 20 min, drink less water in response to a test injection of AngII than do vehicle-treated controls given the same test injection. The present studies were designed to test three potential explanations for the reduced dipsogenic potency of AngII after repeated administration. To this end, we tested for motor impairment caused by repeated injections of AngII, for a possible role of visceral distress or illness, and for differences in the pressor response to the final test injection of AngII. We found that repeated injections of AngII neither affected drinking stimulated by carbachol nor did they produce a conditioned flavor avoidance. Furthermore, we found no evidence that differences in the pressor response to the final test injection of AngII accounted for the difference in intake. In light of these findings, we are able to reject these three explanations for the observed behavioral desensitization, and, we suggest instead that the mechanism for this phenomenon may be at the level of the receptor.