Modification of Oxalis tuberosa starch with OSA, characterization and application in food-grade Pickering emulsions.

The emulsifying properties of Oxalis tuberosa starch (native and chemically modified) were evaluated in Pickering emulsions based on the emulsification index, emulsion stability over time and emulsion morphology. The best conditions of chemical modification were found by esterification of starch with octenyl succinic anhydride (OSA) at a concentration of 3% and a reaction time of 2 h, achieving a degree of substitution of 0.033 ± 0.001. The results obtained using Fourier-transform infrared spectroscopy, a Rapid Visco Analyzer, and differential scanning calorimetry, indicated that the starch underwent a change in its structure and that the insertion of the OSA groups was achieved. The amphipathic characteristics of OSA starch were evaluated by forming oil-in-water emulsions. Various concentrations of OSA-starch granules (1, 2.5 and 5 wt%) were used. A higher concentration of particles produced a smaller droplet size of emulsions (76.5 ± 0.9 µm) compared to those formed at a lower concentration of 1% (92.5 ± 1.0 µm). Therefore, the starch modified with OSA displayed the necessary characteristics to be adsorbed at the oil-water interface, achieving Pickering emulsion stabilization.

The use of a D-optimal design to model the effects of temperature, NaCl, type and acid concentration on Lactobacillus pentosus IGLAC01.

AIMS: To study the effects of temperature, NaCl and acid (HCl, citric, acetic and lactic) concentrations on the specific growth rate (mu), lag phase (lambda), and h0 of Lactobacillus pentosus IGLAC01. METHODS AND RESULTS: Response surface (RS) methodology (D-optimal design) was used with a dummy variable, to account for the different types of acids. The variable ranges were: 16-30 degrees C, 0-70 g l-1 NaCl, and 0-5 g l-1 acid (or 0-2.5 g l-1 HCl). Time to detection from optical density data was used to deduce mu and lambda. The RS models for log2mu and log2lambda, according to acid types, were estimated and the effects of variables were quantified by their z-generalized values. A relationship between ln h0 with temperature was also found. CONCLUSIONS: The mu of L. pentosus IGLAC01 can be doubled by increasing temperature by 10.3 degrees C or by decreasing NaCl by 48 g l-1 (harmonic, averaged, z values, Z); citric was the least inhibitory acid (zmu=-96.2) and lactic the strongest (zmu=-5.7), according to their generalized z values, z. A twofold lambda increase was achieved from a decrease of 3.1 degrees C (HCl), or 4.27 degrees C (citric) or 36 g l-1 NaCl increase (both acids) (expressed as zlambda ); the same effect was obtained from a decrease of 4.37 degrees C, 54 g l-1 NaCl increase, or 10 g l-1 acetic or lactic acid additions (expressed as Zlambda values). SIGNIFICANCE AND IMPACT OF THE STUDY: Valuable information on the effects of environmental variables on the biological parameters of L. pentosus IGLAC01, which could be used for the optimization of olive, cucumber or other vegetable fermentations, is obtained.

Gracilis muscle neosphincter for treating urinary incontinence.

The purpose of this study was to test the anatomical and functional feasibility of using a gracilis muscle free flap to create a urinary sphincter. Anatomical studies were performed in 12 human cadavers and short-term (n = 7) and long-term (n = 8) functional studies were performed in dogs. In the short-term functional studies, the left gracilis muscle was transferred into the pelvis and wrapped around the urethra and the right gracilis muscle was wrapped around a stent. A cuff electrode was placed on the muscle's nerve pedicle and used to stimulate the neosphincter while peak pressure, fatigue rate, and perfusion measurements were performed. In the long-term functional studies, intramuscular electrodes were inserted into the neosphincter to stimulate the flap. The flaps were wrapped around the urethra and dogs were followed for 16 weeks, during which time urodynamic measurements were performed. Our anatomical studies demonstrated that the gracilis muscle free flap could be transferred into the pelvis to create a urinary neosphincter. Our short-term functional study demonstrated that gracilis muscle free-flap function and perfusion were not compromised by transfer. In our long-term functional study, all neosphincters provided bladder outlet resistance pressures consistent with continence. Our anatomical, short-term, and long-term functional studies indicate that a gracilis muscle free-flap neosphincter is an effective procedure for treating urinary incontinence.

Synthesis and pharmacological evaluation of 2, 3-dihydro-3-oxo-4H-thieno[3,4-e][1,2,4]thiadiazine 1,1-dioxides as voltage-dependent calcium channel blockers.

The synthesis of a novel series of 2,3-dihydro-3-oxo-4H-thieno[3, 4-e][1,2,4]thiadiazine 1,1-dioxides and their pharmacological evaluation as drugs with effects on the rat cardiovascular system are described. The compounds under study were synthesized via Curtius rearrangement of appropriate sulfamoylacylazides which, in turn, were prepared from known starting materials. In isolated rat portal vein, these thienothiadiazines, like verapamil and diazoxide, inhibited the spontaneous motility produced by KCl (20 mM). In addition, the new compounds, like verapamil and unlike diazoxide, also exhibited inhibitory effects in the same preparation when the cell membrane was depolarized by an increased extracellular KCl concentration (80 mM) and, consequently, the membrane potential approached a level close to the K(+) equilibrium potential. Further characterization of this inhibitory activity led to the identification of a selective inhibitory effect of the new compounds on KCl (80 mM)-induced 45Ca(2+) uptake in the same vascular tissue. When tested in vivo (anaesthetized normotensive rats), acute administration of verapamil, diazoxide and some of the most in vitro potent compounds in 45Ca(2+) uptake experiments produced a gradual, dose-dependent and sustained decrease in diastolic arterial blood pressure, devoid of cardiac effects. These results suggest that, like verapamil, the cardiovascular effects produced by the new thienothiadiazines seem to be due, at least in part, to a blockade of transmembrane voltage-dependent calcium channels present in vascular smooth muscle cells and not to an activation of ATP-sensitive K(+) channels. Compounds 5b, 5e and 5i have been selected for further studies as antihypertensive agents.

Synthesis and biological evaluation of 1,3,4-triaryl-3-pyrrolin-2-ones, a new class of selective cyclooxygenase-2 inhibitors.

The synthesis and structure activity relationships (SAR) of a series of novel selective COX-2 inhibitors are reported. The results show that some of the 1,3,4-triaryl-3-pyrrolin-2-ones 1 are more potent as COX-2 inhibitors than celecoxib, and that lactam Id has the same selectivity.

Functional profile of almotriptan in animal models predictive of antimigraine activity.

Almotriptan is a new 5-HT(1B/1D) receptor agonist whose clinical efficacy for the treatment of migraine attacks has been demonstrated in Phase III clinical trials. We now compare the functional profile of almotriptan (assessed using animal models) with that of sumatriptan. Almotriptan selectively increased carotid vascular resistance in anaesthetised cats after intravenous or intraduodenal administration (ED(100)=11 microg/kg, i.v.; ED(50)=339 microg/kg, i. d.) and in anaesthetised beagle dogs following intravenous administration (ED(50)=116 microg/kg). A study in anaesthetised cats also demonstrated that almotriptan acts by selectively increasing the resistance of the carotid arteriovenous anastomoses without adversely affecting brain irrigation. In addition, almotriptan inhibited meningeal extravasation produced by electrical stimulation of the trigeminal ganglion in anaesthetised guinea pigs in the dose range of 0.3-3 mg/kg, i.v. In conclusion, almotriptan is both a selective constrictor affecting intracranial blood vessels and an inhibitor of neurogenically evoked plasma protein extravasation of the dura mater.

Effects of rolipram on the elevated plus-maze test in rats: a preliminary study.

The objective of the present study was to assess the behavioural effects of rolipram, a specific cAMP phosphodiesterase (PDE4) inhibitor, in the elevated plus-maze (EPM) test in rats. Results showed that rolipram at the highest dose tested (0.1 mg/kg) increased the percentage of both time spent and entries into open arms, although a decrease of locomotor activity in the EPM test was also observed. In contrast, diazepam (3.0 mg/kg) exhibited the typical profile of an anxiolytic in the EPM test, increasing the percentage of time spent and entries into open arms as well as locomotor activity. A posterior statistical analysis, however, established that the effects of both rolipram and diazepam on parameters denoting anxiolytic-like activity were statistically independent from those reflecting locomotor activity reduction. Furthermore, the effects of both rolipram and diazepam were shown to be distinct from those exhibited by tricyclic antidepressant imipramine which did not show any anxiolytic-like effects in the EPM test, although a reduction of locomotor activity was also detected. Although these preliminary results suggest that rolipram may have some anxiolytic-like properties on the EPM test in rats, such an interpretation should be taken cautiously due to the observed effects on locomotor activity, which could complicate the interpretation of results from rolipram and other PDE4 inhibitors in the current test and in other anxiety animal models.

Preliminary evidence for an involvement of the cholinergic system in the sedative effects of rolipram in rats.

Rolipram is a specific cAMP phosphodiesterase type 4 (PDE4) inhibitor in the brain, which induces an increase in the intracellular levels of cAMP. Rolipram produces characteristic alterations in animal behavior, which have been suggested to be mediated mainly through an intracellular mechanism involving an increase in cAMP. However, specific mechanisms mediating the sedative effects of this compound have not yet been investigated. Because several lines of evidence indicate that the acetylcholine neural system may be involved in some effects of PDE4 inhibitors, the aim of this study was to elucidate whether the neurotransmitter acetylcholine is involved in the sedative effects induced by rolipram. The present study assessed the motor effects of rolipram in an exploratory behavioral test, the open field, in Wistar rats. The results show that rolipram (0.1-3.0 mg/kg SC) induced potent and dose-dependent hypoactivity, decreasing both locomotion and rearing. Physostigmine (0.03-0.3 mg/kg SC) potentiated a subeffective dose of rolipram (0.03 mg/kg SC), resulting in strong sedation, similar to that following higher doses of either rolipram or physostigmine alone, whereas the reduction in locomotor activity induced by rolipram (0.3 mg/kg SC) was completely reversed by scopolamine (0.03-0.3 mg/kg SC). These data provide preliminary evidence suggesting the involvement of the acetylcholinergic system in the sedative effects of rolipram.

In vivo control of 5-hydroxytryptamine release by terminal autoreceptors in rat brain areas differentially innervated by the dorsal and median raphe nuclei.

Selective serotonin reuptake inhibitors (SSRIs) reduce the 5-HT release in vivo. This effect is due to the activation of somatodendritic 5-HT1A receptors and it displays a regional pattern comparable to that of selective 5-HT1A agonists, i.e., preferentially in forebrain areas innervated by the dorsal raphe nucleus (DRN). However, despite a comparatively lower 5-HT1A-mediated inhibition of 5-HT release and a greater density of serotonergic uptake sites in hippocampus, the net elevation produced by the systemic administration of SSRIs is similar in various forebrain areas, regardless of the origin of serotonergic fibres. As terminal autoreceptors may also limit the SSRI-induced elevations of 5-HT in the extracellular brain space, we reasoned that a differential control of 5-HT release by terminal autoreceptors in DRN- and median raphe-innervated areas might be accountable. To examine this possibility, we have conducted a regional microdialysis study in the DRN, MRN and four forebrain regions preferentially innervated either by the DRN (frontal cortex, striatum) or the median raphe nucleus (MRN; dorsal and ventral hippocampus) using freely moving rats. Dialysis probes were perfused with 1 microM of the SSRI citalopram to augment the endogenous tone on terminal 5-HT autoreceptors. The non-selective 5-HT1 antagonist methiothepin (10 and 100 microM, dissolved in the dialysis fluid) increased extracellular 5-HT in frontal cortex and dorsal hippocampus in a concentration-dependent manner. The 5-HT(1B/1D) antagonist GR 127935 was ineffective at 10 microM and tended to reduce 5-HT in dorsal hippocampus at 100 microM. The local infusion of 100 microM methiothepin significantly elevated the extracellular 5-HT concentration to 142-173% of baseline (mean values of 260 min post-administration) in the DRN, MRN, frontal cortex, striatum and hippocampus (dorsal and ventral). Comparable elevations were noted in the four forebrain regions examined. As observed in frontal cortex and dorsal hippocampus, the perfusion of 10 microM GR 127935 did not elevate 5-HT in DRN. MRN, striatum or ventral hippocampus. Because the stimulated 5-HT release in the DRN has been suggested to be under control of 5-HT(1B/1D) receptors, we examined the possible contribution of these receptor subtypes to the effects of methiothepin in the DRN. The perfusion of sumatriptan (0.01-10 microM) or GR 127935 (0.01-10 microM) did not significantly modify the 5-HT concentration in dialysates from the DRN. Thus, the present data suggest that the comparable effects of SSRIs in DRN- and MRN-innervated forebrain regions are not explained by a preferential attenuation of 5-HT release by terminal 5-HT1B autoreceptors in hippocampus, an area with a low inhibitory influence of somatodendritic 5-HT1A receptors. Methiothepin-sensitive autoreceptors (possibly 5-HT1B) appear to play an important role not only in the projection areas but also with respect to the control of 5-HT release in the DRN and MRN. In addition, our findings indicate that GR 127935 is not an effective antagonist of the actions of 5-HT at rat terminal autoreceptors.

Comparison of effects of a range of 5-HT receptor modulators on consumption and preference for a sweetened ethanol solution in rats.

This study assessed the effects of a range of serotonergic agents on preference for a slightly sweetened ethanol solution (10% ethanol, 3% glucose) in rats. A two-bottle, free-choice paradigm was used following induction of ethanol consumption. The model used provides a robust and reliable level of ethanol self-administration in normal laboratory rats. Ethanol consumption was significantly and selectively reduced by the 5-hydroxytryptamine-1A (5-HT1A) full agonist 8-OH-DPAT (0.3-1.0 mg/kg) and the 5-HT3 antagonist granisetron (0.1-1.0 mg/kg). Non-specific reductions in fluid consumption were induced by the 5-HT1B agonist RU 24969 (0.1-1.0mg/kg) and the 5-HT2 antagonist ritanserin (1.0-6.0 mg/kg). These studies thus confirm the potential for decreasing ethanol consumption and ethanol preference of 5-HT1A agonists and 5-HT3 antagonists, but failed to find any selective effects for agents acting at 5-HT1B or 5-HT2 receptors.

CCK is involved in both peripheral and central mechanisms controlling food intake in chickens.

The aim of this work was to study the involvement of cholecystokinin (CCK) in the control of food intake in chickens. The following aspects were studied: 1) the effects of intravenous and intracerebroventricular sulfated octapeptide of CCK (CCK-8s) on voluntary food intake; 2) the effects of two CCK-receptor antagonists. L-365,260 and L-364,718, on food intake; and 3) the ability of such drugs to block the effects of CCK-8s on food intake in the chicken. Intravenous and intracerebroventricular CCK-8s caused a decrease in food intake. Intraperitoneal L-365,260, a CCK-receptor antagonist with low affinity for the two CCK receptors described in the chicken, increases food intake. Intracerebroventricular L-364,718, a drug that has high affinity for the chicken central CCK-receptor type, increased food intake. The effect of intravenous CCK-8s on food intake was not blocked by L-364,718 or L-365,260, whereas that of intracerebroventricular CCK-8s was blocked by intracerebroventricular L-364,718. It is concluded that central endogenous CCK plays a role in the control of food intake, which is dependent on central CCK-receptor type; nevertheless, peripheral CCK also decreases food intake acting on the peripheral CCK-receptor type. The fact that intracerebroventricular L-364,718 is able to increase food intake is related to its high affinity for the central CCK-receptor type of this species. Finally, three different speculations that might explain the fact that intraperitoneal L-365,260 increases food intake are discussed.

Characterization of 5-HT receptors on human pulmonary artery and vein: functional and binding studies.

1. This study aimed to investigate the 5-hydroxytryptamine (5-HT) receptors mediating contraction of ring preparations isolated from human pulmonary arteries and veins. In functional studies, the responses to 5-HT, sumatriptan, ergotamine, serotonin-O-carboxymethyl-glycyl-tyrosinamide (SCMGT), alpha-methyl 5-HT (alpha-Me) and 2-methyl 5-HT (2-Me) were studied with WAY100635, GR127935, ritanserin, zacopride and SB204070 as antagonists. 2. All agonists produced concentration-dependent contractions of human pulmonary artery and vein preparations. The order of potency (-log ECS0 values) was ergotamine (6.88) > 5-HT (6.41) > or = SCMGT (6.20) = sumatriptan (6.19) > or = alpha-Me (6.04) in the artery, and ergotamine (7.84) > 5-HT (6.96) > sumatriptan (6.60) = alpha-Me (6.56) > SCMGT (6.09) in the vein. The potency of each agonist, except for SCMGT, was greater in vein than in artery preparations. Contractile responses to 5-HT were similar in intact and endothelium-denuded preparations but responses to sumatriptan were enhanced in artery rings without endothelium. 3. GR127935 (1 nM to 0.5 microM) produced an unsurmountable antagonism of the response to 5-HT, sumatriptan, ergotamine and SCMGT. Ritanserin (1 nM to 1 microM) also reduced the maximum contractile responses to 5-HT, ergotamine and alpha-Me in artery and vein preparations without affecting those to sumatriptan and SCMGT. In endothelium-denuded preparations, surmountable antagonism of sumatriptan by GR127935 (in the presence of ritanserin) and of alpha-Me by ritanserin (in the presence of GR127935) allowed for the calculation of the apparent pK(B) values of GR127935 (9.17+/-0.11 in artery and 9.11+/-0.05 in vein) and ritanserin (8.82+/-0.09 in artery and 8.98+/-0.12 in vein). 4. WAY100635 (1 nM to 1 microM), zacopride (1 nM to 1 microM), or SB204070 (1 nM) did not significantly alter the concentration-response curves for 5-HT, sumatriptan, ergotamine, SCMGT or 2-Me in human pulmonary artery or vein thus indicating that 5-HT1A, 5-HT3 and 5-HT4 receptors are presumably not involved in the contractile response to these agonists. 5. Binding studies using selective radioligands for different 5-HT receptors could not detect the presence of 5-HT1A receptor binding in human pulmonary blood vessels whereas the 5-HT(1B/1D) radioligand [3H]-5CT significantly labelled a population of specific binding sites in both vessel types. The presence of 5-HT2A receptors could also be inferred from the level of binding of [3H]-ketanserin to membranes obtained from human pulmonary vessels, although significance could not be reached for arteries. 5-HT4 specific receptor binding was scarce in veins and absent in the case of arteries. 6. These findings indicate that the human pulmonary artery and vein have a mixed functional population of 5-HT(1B/1D) and 5-HT2A receptors mediating the contractile response to 5-HT which is consistent with results of the binding studies.

Effects of a range of dopamine receptor agonists and antagonists on ethanol intake in the rat.

The aim of this study was to assess the effects of a range of dopaminergic agents on consumption of an ethanol solution (10% ethanol, 3% glucose) in rats. A two-bottle, free-choice paradigm was used following induction of ethanol consumption and preference in standard laboratory rats. The model used provides a robust and reliable level of ethanol oral administration in normal laboratory rats. Both ethanol intake and preference were reduced by a dopamine D1 receptor partial agonist, SFK 38393 ((+/-)-1-phenyl-2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8-diol hydrochloride), in a dose-dependent manner. The dopamine D2/D3 receptor agonist 7-OH-DPAT ((+/-)-7-hydroxy-N,N-(di-n-propyl-2-aminotetralin)) at the lowest dose of 0.01 mg/kg increased both ethanol intake and preference. At higher doses (0.03-0.1 mg/kg) no significant effects were found. The dopamine D1 receptor antagonist SCH 23390 (R-(+)-7-chloro-2,3,4,5-tetrahydro-3-methyl-1-phenyl-1H-3-benzazepine-8- ol), dopamine D2/D3 receptor antagonist raclopride and 5-HT2/D2 receptor antagonist risperidone did not affect ethanol consumption, although all at high doses induced a significant decrease in water intake, indicating a non-specific decrease in consummatory behavior with these compounds. These results suggest the involvement of the dopaminergic system in ethanol intake and ethanol reinforcement with dopamine D1 and D2/D3 receptors playing opposing roles. Blockade of dopamine D2 receptors had no selective effect on ethanol consumption and ethanol preference.

Effects of 5-HT4 receptor antagonists on rat behaviour in the elevated plus-maze test.

The anxiolytic-like effects of a variety of 5-HT receptor agonists and antagonists have been intensively studied in animal models. However, no direct effects of agents modulating 5-HT4 receptors have been reported, in spite of their suggestive location in the brain. The objective of the present study was the determination of the effects of two selective 5-HT4 receptor antagonists, SB 204070 [1-butyl-4-piperidinylmethyl)-8-amino-7-chloro-1,4-benzodioxan-5-carboxylate] and GR 113808 [[1-[2-methylsulphonyl)amino]ethyl]-4-piperidinyl]methyl 1-methyl-1 H-indole-3-carboxylate], in the elevated plus-maze test in rats. Results have shown that both 5-HT4 receptor antagonists exhibit an anxiolytic-like profile, although only at the dose of 1.0 mg/kg (s.c.). At this dose, both compounds significantly increased the percentage of time spent in open arms exploration, while other variables evaluated remained unaffected at the dose range tested. Results suggest that 5-HT4 receptor antagonists could have some anxiolytic-like properties, although their effects seem more limited and less consistent than those presented by classic anxiolytics, such as diazepam. However, they are similar to those exhibited by granisetron [endo-1-methyl-N-(9-methyl-9-azabicyclo[3.3.1]non-3-yl)-1 H-indazole-3-carboxamide], a 5-HT3 receptor antagonist.

GR 127935 blocks the locomotor and antidepressant-like effects of RU 24969 and the action of antidepressants in the mouse tail suspension test.

The 5-HT1A/B agonist RU 24969 induces hyperactivity in rodents and also shows antidepressant-like effects in some animal models of depression. We have examined the effects of selective antagonists at 5-HT1A and 5-HT1B/D receptors (WAY 100135 and GR 127935, respectively) on both the hyperlocomotor and anti-immobility effects of RU 24969. While a high dose of WAY 100135 (10 mg/kg) had no effect in either paradigm, GR 127935 attenuated the behavioural effects of RU 24969 in both. WAY 100135 was also without effect on the antidepressant effect of paroxetine, while GR 127935 blocked the effects of paroxetine (1 mg/kg) and imipramine (10 mg/kg). Furthermore, while coadministration of paroxetine or imipramine enhanced the effects of RU 24969 in the mouse tail suspension test, imipramine had no effect on the locomotor activating effects of the 5-HT1B agonist, suggesting different neural substrates may underly the effects in the different tests. These studies indicate a role for the 5-HT1B/D receptor in the mediation of the effects of antidepressant treatment.

L-364,718 and L-365,260, two CCK antagonists, have no affinity for central benzodiazepine binding sites in chickens.

It has recently been demonstrated that L-365,260, a CCK-B antagonist in mammals, causes an increase in food intake in chickens. In contrast, L-364, 718, a CCK-A antagonist in mammals, shows this effect only at very high dose levels. It has been shown that L-365,260 has very low affinity for chicken CCK receptors. Thus, the mechanism of action of L-365,260 remains unknown. As L-365,260 is a benzodiazepine derivative, one may hypothesize that it would be acting on benzodiazepine binding sites. The aims of this work were to establish the existence of benzodiazepine binding sites in the chicken brain, and to check the possibility that L-365,260 was acting on these receptors, determining the affinity of L-364,718 and L-365,260 for them. We have found specific binding for tritiated flunitrazepam (a benzodiazepine agonist) ([3H]-flunitrazepam) in chicken brain membranes. A single binding site was detected with a Kd of 3.58 +/- 0.97 nM and a Bmax of 451.6 +/- 23.3 fmol/mg protein L-365,260 and L-364,718 exhibited very low affinity for these binding sites (Ki = 1.17 x 10(-6) +/- 0.16 x 10(-6) M and Ki > 10(-5) M, respectively). Thus, these results demonstrate that the increase in food intake caused by L-365,260 in the chicken is not due to a direct action on benzodiazepine receptors. Other possible explanations for its effect are discussed.

Central and peripheral cholecystokinin receptors in chickens differ from those in mammals.

Specific binding for the radioligand [3H]CCK-8s has been identified in chicken brain, hypothalamus, pancreas, gallbladder and caecum membranes. This binding was found to be of high affinity, low capacity and saturable, suggesting the presence of specific CCK receptors in these tissues. Scatchard analysis indicated the existence of a single binding site for each tissue. Dissociation constant (kd) values were 0.63 +/- 0.18, 0.73 +/- 0.13, 0.85 +/- 0.12, 1.47 +/- 0.21 and 0.96 nM for brain, hypothalamus, pancreas, caecum and gallbladder, respectively. Binding densities (Bmax) were higher for brain, pancreas and caecum (32.60 +/- 10.70, 30.33 +/- 2.40 and 35.83 +/- 5.10 fmol/mg protein, respectively) than for the other two tissues (9.75 +/- 1.90 and 6.31 fmol/mg protein for hypothalamus and gallbladder, respectively). As in mammals, CCK-4 shows high affinity for CCK receptors located in chicken brain and hypothalamus, and very low affinity for those located in peripheral structures. L-364,718 (a CCK-A antagonist) showed a relative selectivity and a high affinity for those receptors located in central tissues, whereas L-365,260 (a CCK-B antagonist) is almost inactive in all studied tissues. These results give support for the existence of at least two distinct CCK receptors in birds and that these receptors are relatively different from those described in mammals.