Dopamine mediation of the feeding response to violations of spatial and temporal expectancies.

The present studies were aimed at further characterizing the role of DA in motivation. Rats, conditioned to expect food in one environment and no food in another, all received food on the test night. Those in the environment in which food was unexpected ate four times as much as those eating where food was expected. The overeating was eliminated by administration of the D2 antagonist raclopride. Another expectancy, timing of light offset in rats entrained to a fixed light--dark cycle, was violated by unexpectedly turning the lights off 1 h early. This provoked an elevation in food intake, which was also eliminated by the administration of raclopride. Feeding in two other situations not involving violation of expectancies (food deprivation; normal light offset) was unaffected by DA antagonism. These findings support the idea that DA signals errors in expectancy and that DA signaling is necessary for certain behavioral responses to unexpected events.

NaCl detection thresholds: comparison of Fischer 344 and Wistar rats.

Adult Fischer 344 (F344) rats fail to display any preference for NaCl solutions at concentrations typically preferred by other rat strains. To determine whether this behavior is due to a strain difference in NaCl detection threshold, a conditioned taste aversion (CTA) was first established to a suprathreshold concentration of NaCl (0.1 M). Then, a series of dilute NaCl solutions, ranging from 0.0 to 0.011 M NaCl, were presented to F344 (n = 16) and Wistar (n = 16) rats. The lowest concentration at which there was a reliable difference in the preference scores of conditioned and control rats was defined as the detection threshold. Results indicate that the detection threshold for NaCl lies between 0.001 and 0.002 M NaCl for both F344 and Wistar rats. The addition of the sodium channel blocker amiloride to the NaCl solutions raised the detection threshold 10-fold to 0.03-0.04 M NaCl for both strains of rats. These results suggest that the NaCl detection thresholds of F344 and Wistar rats are similar and that these strains do not differ in the degree to which amiloride raises this threshold.

NaCl preference increases during pregnancy and lactation: assessment using brief access tests.

Pregnancy and lactation are characterized by increases in NaCl intake, as determined by long-term consumption tests, which cannot examine the relative contribution of taste and postingestive factors to this phenomenon. Consequently, in this study, changes in NaCl preference during pregnancy and lactation were studied in nulliparous Long-Evans rats using a brief access test (lickometer). In Experiment 1, rats were maintained on a Na(+)-adequate diet (0.03% Na(+)), habituated to lickometer testing, and subsequently assessed during pregnancy and lactation with three 30-s exposures to each of seven taste solutions: 0.075 M sucrose (base), 0.089 M NaCl in base, 0.158 M NaCl in base, 0.281 M NaCl in base, 0.5 M NaCl in base, 0.158 M NaCl and 0.281 M NaCl. Results indicated higher lick rates to the 0.5 M NaCl in base, 0.158 M NaCl and 0.281 M NaCl solutions during late pregnancy and late lactation (Day 13 and beyond). In Experiment 2, a comparison of two diets differing in sodium content (0.03% vs. 0.3% Na(+)) determined that these changes in NaCl preference during pregnancy and lactation were unrelated to dietary sodium. Thus, the apparent increase in NaCl preference during pregnancy and lactation, independent of dietary sodium, suggests that this change in preference is not in response to physiological sodium need.

Amiloride-sensitive signals and NaCl preference and appetite: a lick-rate analysis.

Rats prefer hypotonic and isotonic NaCl solutions to water in long-access drinking paradigms. To focus on the role of taste signals in NaCl preference, licking patterns of rats with 30-s exposure to NaCl solutions (0-0.5 M) were examined when they were either water deprived, sodium depleted, or not deprived (NaCl mixed in dilute sucrose). In all three conditions, rats displayed a preference for NaCl. The addition of 100 microM amiloride, a sodium channel blocker, to NaCl did not change rats' licking when they were sodium replete but dramatically reduced licking when they were deplete. Transection of the chorda tympani (CT) nerve, an afferent pathway for amiloride-sensitive Na(+) signals, had no effect on NaCl preference in nondeprived rats and only a modest effect on those that were Na(+) deplete. Amiloride was found to exert significant suppression of NaCl intake in Na(+)-depleted rats with transection of the CT, supporting the existence of other afferent pathways for transmission of amiloride-sensitive Na(+) signalling. Together, these studies argue for the involvement of different neural signalling mechanisms in NaCl preference in the presence and absence of explicit Na(+) need.

Neonatal chorda tympani transection permanently disrupts fungiform taste bud and papilla structure in the rat.

The present report examined the morphology of fungiform papillae in adult rats that received bilateral chorda tympani transection at 10 days of age. Tongue tissue was examined using surface-structure analysis. Counts were made of fungiform papillae with a pore, fungiform papillae with no pore and fungiform papillae with a keratinized conical surface; a feature referred to as "filiform-like. " Neonatal chorda tympani nerve transection resulted not only in a loss of taste buds but also in a permanent loss in numbers of fungiform papillae. Compared with an average of 152 fungiform papillae in sham-operated control rats, there was an average of only 54 fungiform papillae after neonatal chorda tympani transection. Nearly 80% of these fungiform papillae in neonatal chorda tympani transected rats were filiform-like. No filiform-like papillae were noted in sham-operated rats. These results suggest that the chorda tympani nerve is necessary during an early postnatal period of development to maintain normal fungiform papillae morphology.

Ethanol-induced c-fos expression in catecholamine- and neuropeptide Y-producing neurons in rat brainstem.

BACKGROUND: Previous studies have used c-Fos-like immunoreactivity (cFLI) to examine the neuroanatomical location of cells that are activated in response to ethanol administration. However, the use of cFLI alone fails to reveal the phenotypical identity of cells. In the present study we used double-labeling procedures to identify the neurochemical phenotype of neurons that showed ethanol-induced cFLI in the rat brainstem. METHODS: Individual groups of rats received intraperitoneal injection of ethanol (1.5 g/kg or 3.5 g/kg) or isotonic saline (23 ml/kg). To assess the specificity of cFLI induced by ethanol, we injected other rats with the drug lithium chloride (LiCl; 76 mg/kg). Two hours after injection, rats were killed and their brains were processed for immunohistochemistry. RESULTS: Both doses of ethanol promoted cFLI in several brainstem regions, including the nucleus of the solitary tract (NTS), the locus coeruleus (LC), and the ventrolateral medulla (VLM). Although LiCl caused significant cFLI in the NTS, this drug promoted only minimal cFLI in the VLM and no significant activation in the LC. We found that a significant proportion of tyrosine hydroxylase (TH)-positive neurons coexpressed ethanol-induced cFLI in the VLM (approximately 75-85%), the NTS (approximately 65-75%), and the LC (approximately 30-65%). Additionally, a significant proportion of neuropeptide Y (NPY)-producing neurons in the VLM coexpressed ethanol-induced cFLI (approximately 60-75%). On the other hand, LiCl promoted activation of TH-positive neurons in the VLM and the NTS but failed to stimulate cFLI in TH-producing neurons in the LC or in NPY-producing neurons of the VLM. CONCLUSIONS: Neurons in the rat brainstem that show ethanol-induced c-Fos expression produce catecholamines and NPY. This research demonstrates the usefulness of double-labeling immunohistochemistry procedures for identifying the neurochemical identity of neurons that are activated after ethanol administration.

High ethanol consumption and low sensitivity to ethanol-induced sedation in protein kinase A-mutant mice.

Both in vitro and in vivo evidence indicate that cAMP-dependent protein kinase (PKA) mediates some of the acute and chronic cellular responses to alcohol. However, it is unclear whether PKA regulates voluntary alcohol consumption. We therefore studied alcohol consumption by mice that completely lack the regulatory IIbeta (RIIbeta) subunit of PKA as a result of targeted gene disruption. Here we report that RIIbeta knockout mice (RIIbeta-/-) showed incr eased consumption of solutions containing 6, 10, and 20% (v/v) ethanol when compared with wild-type mice (RIIbeta+/+). On the other hand, RIIbeta-/- mice showed normal consumption of solutions containing either sucrose or quinine. When compared with wild-type mice, the RIIbeta-/- mice were found to be less sensitive to the sedative effects of ethanol as measured by more rapid recovery from ethanol-induced sleep, even though plasma ethanol concentrations did not differ significantly from those of controls. Finally, both RIbeta- and catylatic subunit beta1-deficient mice showed normal voluntary consumption of ethanol, indicating that increased ethanol consumption is not a general characteristic associated with deletion of PKA subunits. These data demonstrate a role for the RIIbeta subunit of PKA in regulating voluntary consumption of alcohol and sensitivity to the intoxication effects that are produced by this drug.

The induction of c-Fos in the NTS after taste aversion learning is not correlated with measures of conditioned fear.

The induction of c-Fos-like immunoreactivity (c-FLI) in the nucleus of the solitary tract (NTS) has been shown to be correlated with behavioral expression of a conditioned taste aversion (CTA). However, because this cellular response is also dependent on an intact amygdala, it may represent the activation of a stress-related autonomic response. The present experiments addressed this possibility by evaluating the correlation between c-FLI in the intermediate division of the NTS (iNTS) and 2 measures of conditioned fear: freezing and changes in mean arterial pressure (MAP) and heart rate (HR). Exposure to the taste conditioned stimulus (CS) resulted in a marked induction of c-FLI in the iNTS, whereas exposure to a fear CS did not. Further, exposure to a taste CS did not selectively lead to increases in MAP or HR. Results suggest that induction of c-FLI in the iNTS may reflect the activation of a cell population whose function is unique to the CTA paradigm.

c-Fos induction in response to saccharin after taste aversion learning depends on conditioning method.

Increases in c-Fos-like immunoreactivity (FLI) in the intermediate nucleus of the solitary tract (iNTS) have been seen consistently as a correlate of the expression of a conditioned taste aversion (CTA) when conditioning occurs using taste delivery through intraoral (I/O) infusions. The present study examined whether a similar FLI response would occur when conditioning was accomplished by presenting the taste solution in a bottle. I/O and bottle methods generated aversions that were comparable, when judged by the behavioral response of solution rejection. However, elevations in FLI were seen only in animals conditioned with the I/O method. This finding adds to evidence that the neural pathways underlying CTA learning differ as a function of the type of conditioning method used.

Is the use of benzalkonium chloride as a preservative for nasal formulations a safety concern? A cautionary note based on compromised mucociliary transport.

BACKGROUND: Topical nasal solution and suspension delivery systems are available for short- and long-acting vasoconstrictors, ipratropium, cromolyn, azelastine, and glucocorticosteroids. The use of intranasal glucocorticosteroids has increased substantially because the efficacy of these agents has been well established for the treatment of perennial and seasonal allergic rhinitis. Adverse local effects of burning, irritation, and dryness are occasionally associated with glucocorticosteroid nasal preparations. Benzalkonium chloride (BKC) is a quaternary ammonium antimicrobial agent included in some nasal solutions (including glucocorticosteroids) to prevent the growth of bacteria. Some reports suggest that BKC in nasal sprays may cause adverse effects, including reduced mucociliary transport, rhinitis medicamentosa, and neutrophil dysfunction. OBJECTIVE: This article summarizes recent literature about possible adverse biologic effects associated with BKC as a nasal spray preservative by examining its effects on the following properties of mucociliary transport: ciliary motion, ciliary form, ciliary beat frequency, electron microscopy, and particle movement/saccharin clearance tests. CONCLUSION: Both animal and human in vitro data suggest that BKC promotes ciliostasis and reduction in mucociliary transport that may be partially masked by absorption and dilution effects occurring in respiratory mucus. These possible confounding factors may account for several disparate human in vivo results. The use of BKC-free glucocorticosteroid formulations should be considered, particularly in patients who complain of nasal burning, dryness, or irritation.

Neurobiological responses to ethanol in mutant mice lacking neuropeptide Y or the Y5 receptor.

We have previously shown that voluntary ethanol consumption and resistance are inversely related to neuropeptide Y (NPY) levels in NPY-knockout (NPY -/-) and NPY-overexpressing mice. Here we report that NPY -/- mice on a mixed C57BL/6Jx129/SvEv background showed increased sensitivity to locomotor activation caused by intraperitoneal (ip) injection of 1.5 g/kg of ethanol, and were resistant to sedation caused by a 3.5-g/kg dose of ethanol. In contrast, NPY -/- mice on an inbred 129/SvEv background consumed the same amount of ethanol as wild-type (WT) controls at 3%, 6%, and 10% ethanol, but consumed significantly more of a 20% solution. They exhibited normal locomotor activation following a 1.5-g/kg injection of ethanol, and displayed normal sedation in response to 2.5 and 3.0 g/kg of ethanol, suggesting a genetic background effect. Y5 receptor knockout (Y5 -/-) mice on an inbred 129/SvEv background showed normal ethanol-induced locomotor activity and normal voluntary ethanol consumption, but displayed increased sleep time caused by 2.5 and 3.0 g/kg injection of ethanol. These data extend previous results by showing that NPY -/- mice of a mixed C57BL/6Jx129/SvEv background have increased sensitivity to the locomotor activation effect caused by a low dose of ethanol, and that expression of ethanol-related phenotypes are dependent on the genetic background of NPY -/- mice.

cFos induction during conditioned taste aversion expression varies with aversion strength.

Fos-like immunoreactivity (FLI) can indicate the location of neurons activated following expression of conditioned taste aversion (CTA). After one conditioning trial FLI has been identified in the intermediate nucleus of the solitary tract (iNTS) with little expression in other brain regions. The present study assessed the effect of increasing aversion strength on the magnitude and anatomical distribution of FLI during CTA expression. When animals received three rather than one conditioning trial, significant FLI was seen not only in the iNTS but also in the parabrachial nucleus (PBN), and the central nucleus of the amygdala (CNA), regions thought to be important in taste aversion learning.

Insular cortex lesions and taste aversion learning: effects of conditioning method and timing of lesion.

The specific role of insular cortex in acquisition and expression of a conditioned taste aversion was assessed using two different conditioning methods, which vary mode of taste delivery. Involvement of insular cortex in the induction of c-Fos-immunoreactivity in the nucleus of the solitary tract, a cellular correlate of the behavioral expression of a conditioned taste aversion, was also assessed. Electrolytic lesions of insular cortex blocked behavioral expression of a conditioned taste aversion and this was evident not only when lesions were placed prior to conditioning, but also when they were made after conditioning but before testing. In contrast to the effects on behavior, lesions did not completely block the c-Fos-immunoreactivity which accompanies re-exposure to the aversive taste. In addition, the blocking of behavioral evidence of aversion conditioning by cortical lesions was seen both in animals trained under an intraoral acquisition procedure and those trained with bottle-conditioning. This contrasts with previous work with amygdala lesions which showed that amygdala was absolutely necessary for taste aversions conditioned with the intraoral method but not for those conditioned using bottle presentation of the taste. Overall, these findings imply that the details of the neural circuitry involved in taste aversion learning, including its anatomical distribution, complexity and degree of redundancy, vary with the type of conditioning method employed.

Morning sickness and salt intake, food cravings, and food aversions.

Evidence for an association between early pregnancy sickness and offspring salt (NaCl) preference has been obtained from studying offspring as young adults and as infants. To determine whether the association between early pregnancy sickness and salt preference of offspring is secondary to familiar similarity in salt preference, the present study examined the self-reported salt intake and dietary cravings and aversions of pregnant women. Women who reported little or no vomiting (n = 108) were compared to women who reported moderate to severe vomiting (n = 21) during pregnancy. The women's self-reported salt use and reported cravings and aversions for common food were measured via survey for time periods prior to and during their current pregnancy. Women did not differ in reported salt use prior to pregnancy as a function of their pregnancy symptoms. Women reported more aversions during, than prior to, pregnancy (p < 0.05). Women with more severe vomiting reported a greater number of aversions (p < 0.05) both prior to and during pregnancy. There was a significant association between experiencing cravings and aversions prior to pregnancy and experiencing craving and aversions during pregnancy (p < 0.05). These findings do not provide evidence for an association between dietary levels of sodium and the likelihood of experiencing severe pregnancy symptoms. Therefore, these data do not support the suggestion that reported elevations in salt preference in offspring of women with moderate to severe vomiting during pregnancy are mediated by familial dietary practices.

Immune responses in farm workers after exposure to Bacillus thuringiensis pesticides.

Although health risks to pesticides containing Bacillus thuringiensis (Bt) have been minimal, the potential allergenicity of these organisms has not been evaluated. Therefore, a health survey was conducted in farm workers before and after exposure to Bt pesticides. Farm workers who picked vegetables that required Bt pesticide spraying were evaluated before the initial spraying operation (n = 48) and 1 and 4 months after (n = 32 and 20, respectively). Two groups of low- (n = 44) and medium- (n = 34) exposure workers not directly exposed to Bt spraying were also assessed. The investigation included questionnaires, nasal/mouth lavages, ventilatory function assessment, and skin tests to indigenous aeroallergens and to a variety of Bt spore and vegetative preparations. To authenticate exposure to the organism present in the commercial preparation, isolates from lavage specimens were tested for Bt genes by DNA-DNA hybridization. Humoral immunoglobulin G (IgG) and immunoglobulin E (IgE) antibody responses to spore and vegetative Bt extracts were assayed. There was no evidence of occupationally related respiratory symptoms. Positive skin-prick tests to several spore extracts were seen chiefly in exposed workers. In particular, there was a significant (p < 0.05) increase in the number of positive skin tests to spore extracts 1 and 4 months after exposure to Bt spray. The number of positive skin test responses was also significantly higher in high (p < 0.05) than in low- or medium-exposure workers. The majority of nasal lavage cultures from exposed workers was positive for the commercial Bt organism, as demonstrated by specific molecular genetic probes. Specific IgE antibodies were present in more high-exposure workers (p < 0.05) than in the low and medium groups. Specific IgG antibodies occurred more in the high (p < 0.05) than in the low-exposure group. Specific IgG and IgE antibodies to vegetative organisms were present in all groups of workers. Exposure to Bt sprays may lead to allergic skin sensitization and induction of IgE and IgG antibodies, or both.

Amiloride-sensitive sodium signals and salt appetite: multiple gustatory pathways.

In the rat, the ionic specificity of Na+ appetite is thought to rely on amiloride-sensitive Na+ signals conveyed by the chorda tympani (CT) nerve. We evaluated whether robust Na+ appetite relies exclusively on CT-mediated amiloride-sensitive Na+ signals. Amiloride dramatically reduced sham drinking of NaCl (41.9 +/- 9.0 vs. 6.9 +/- 3.7 ml, 0.1 M NaCl without vs. with 100 microM amiloride), which resulted in intake that was not different from intake of a non-Na+ salt solution (8.8 +/- 2.3 ml, 0.15 M KCl). In addition, intake of 0.1 M NaCl in CT-transected (CTX) rats was reduced (35.8 +/- 13.3 vs. 8.67 +/- 3.4 ml, sham-operated vs. CTX rats), but the addition of amiloride (100 microM) further reduced intake in CTX rats (0.5 +/- 0.29 ml). These data support the idea that amiloride-sensitive Na+ channels are the critical gustatory substrate for Na+ identification during Na+ appetite in the rat. However, the data indicate that these amiloride-sensitive signals are not conveyed exclusively by the CT nerve but by an additional afferent pathway.

A cross-sectional survey of sensitization to Aspergillus oryzae-derived lactase in pharmaceutical workers.

BACKGROUND: The presence of IgE-mediated occupational respiratory sensitization to microbial enzymes has been well documented in a variety of industries. Aspergillus oryzae -derived lactase is used as a dietary aid for patients with lactose intolerance. OBJECTIVE: In 1993, a cross-sectional survey of 94 pharmaceutical workers exposed to lactase for a mean duration of 23 months and 24 nonexposed recently hired employees was initiated to identify lactase-sensitized workers and potential risk factors that could be used in making recommendations for preventing future cases of lactase sensitization. METHODS: The survey included a physician-administered questionnaire, skin prick testing to lactase enzyme and a panel of common aeroallergens, and spirometry. RESULTS: Twenty-seven of 94 lactase-exposed workers (29%) had positive skin test responses to lactase. These workers were 9 times more likely to have upper or lower respiratory symptoms compared with workers with negative skin test responses. Atopic workers were 4 times more likely to have lactase skin sensitivity than nonatopic workers. However, atopy was not a risk factor for the development of upper and/or lower respiratory symptoms. Lactase skin reactivity was not observed in the 24 nonexposed employees. CONCLUSION: This cross-sectional survey revealed that atopic workers were more likely to have lactase sensitization and that lactase-sensitized workers were more likely to have upper and/or lower respiratory symptoms, but atopy was not a risk factor for upper or lower respiratory symptoms. In spite of these findings, the company allowed only nonatopic, nonlactase-sensitized workers to continue working in high lactase-exposure areas with careful symptom monitoring and use of protective clothing. Although this strategy was successful in total prevention of new cases of occupational respiratory disease after 5 years, the results of this cross-sectional survey do not support exclusion of atopic workers from working with industrial enzymes.

Food aversion learning: a risk factor for nutritional problems in the elderly?

Food aversions that are acquired as a result of unpleasant experiences with foods represent a potent defense mechanism against poisoning. However, this powerful and durable form of conditioning can also contribute to avoidance of foods that are not poisonous, and are, in fact, quite nutritious. This is because such foods may be coincidentally associated with unpleasant gastrointestinal symptoms, sometimes due to transient, unrelated illness, or unpleasant drug side effects. Most of the studies of naturally occurring learned food aversions in humans have been focused on subjects of college age, so we have limited information about the extent to which such food aversions occur in the elderly. Additionally, most studies have employed questionnaire or interview methods that may have some significant limitations in the accurate assessment of the incidence of food aversions. Thus, although food aversion learning has been thoroughly documented in the animal laboratory, its role in everyday food selection in humans, including the elderly, remains relatively unclear.