Intragastric Research Articles

Hepatoprotective effect of thymol against subchronic toxicity of titanium dioxide nanoparticles: Biochemical and histological evidences

The study was aimed to investigate the protective action of thymol against nano titanium dioxide (nano-TiO2) induced hepatotoxicity in rats. To achieve this purpose, the rats were divided into four groups (n = 6) including control, nano-TiO2 (100 mg/kg), nano-TiO2 + thymol (10 mg/kg) and nano-TiO2 + thymol (30 mg/kg). Intragastric (IG) administration of nano-TiO2 for 60 consecutive days caused widespread histological changes and significantly induced oxidative stress in the liver tissues as manifested by the rise in serum transaminase activities accompanied by marked decline of enzymatic (catalase, superoxide dismutase and glutathione peroxidase) and non-enzymatic (ferric reducing antioxidant power and glutathione) antioxidant levels, and rise of malondialdehyde levels in liver tissue. Pretreatment with thymol (IG) prior to nano-TiO2 administration significantly ameliorated all of biochemical and histopathological alterations in a dose-dependent manner. In conclusion, thymol effectively protects against nano-TiO2-induced hepatotoxicity in rats by its antioxidant properties.

Rewarding Effects of Operant Dry-Licking Behavior on Neuronal Firing in the Nucleus Accumbens Core.

Certain eating behaviors are characterized by a trend of elevated food consumption. However, neural mechanisms mediating the motivation for food consumption are not fully understood. Food impacts the brain-rewarding-system via both oral-sensory and post-ingestive information. Recent studies have reported an important role of visceral gut information in mediating dopamine (DA) release in the brain rewarding system. This is independent of oral sensation, suggesting a role of the gut-brain-DA-axis in feeding behavior. In this study, we investigated the effects of intra-gastric (IG) self-administration of glucose on neuronal firings in the nucleus accumbens (NA) of water-deprived rats. Rats were trained in an operant-licking paradigm. During training, when the light was on for 2 min (light-period), rats were required to lick a spout to acquire the water oral-intake learning, and either an IG self-infusion of 0.4 M glucose (GLU group) or water (H2O group). Rats rested in the dark-period (3 min) following the light-period. Four cycles of the operant-licking paradigm consisting of the light–dark periods were performed per day, for 4 consecutive days. In the test session, the same rats licked the same spout to acquire the IG self-administration of the corresponding solutions, without oral water ingestion (dry licking). Behavioral results indicated IG self-administration of glucose elicits more dry-licking behavior than that of water. Neurophysiological results indicated in the dark period, coefficient of variance (CV) measuring the inter-spike interval variability of putative medial spiny neurons (pMSNs) in the NA was reduced in the H2O group compared to the GLU group, while there was no significant difference in physical behaviors in the dark period between the two groups. Since previous studies reported that DA release increases CV of MSNs, the present results suggest that greater CV of pMSNs in the GLU group reflects greater DA release in the NA and elevated motivation in the GLU group, which might increase lickings in the test session in the GLU group compared to the H2O group.

Saccharomyces boulardii CNCM I-745 supplementation reduces gastrointestinal dysfunction in an animal model of IBS.

BackgroundWe evaluated the effect of Saccharomyces boulardii CNCM I-745 on intestinal neuromuscular anomalies in an IBS-type mouse model of gastrointestinal motor dysfunctions elicited by Herpes Simplex Virus type 1 (HSV-1) exposure.MethodsMice were inoculated intranasally with HSV-1 (102 PFU) or vehicle at time 0 and 4 weeks later by the intragastric (IG) route (108 PFU). Six weeks after IG inoculum, mice were randomly allocated to receive oral gavage with either S. boulardii (107 CFU/day) or vehicle. After 4 weeks the following were determined: a) intestinal motility using fluorescein-isothiocyanate dextran distribution in the gut, fecal pellet expulsion, stool water content, and distal colonic transit of glass beads; b) integrity of the enteric nervous system (ENS) by immunohistochemistry on ileal whole-mount preparations and western blot of protein lysates from ileal longitudinal muscle and myenteric plexus; c) isometric muscle tension with electric field and pharmacological (carbachol) stimulation of ileal segments; and d) intestinal inflammation by levels of tumor necrosis factor α, interleukin(IL)-1β, IL-10 and IL-4.ResultsS. boulardii CNCM I-745 improved HSV-1 induced intestinal dysmotility and alteration of intestinal transit observed ten weeks after IG inoculum of the virus. Also, the probiotic yeast ameliorated the structural alterations of the ENS induced by HSV-1 (i.e., reduced peripherin immunoreactivity and expression, increased glial S100β protein immunoreactivity and neuronal nitric oxide synthase level, reduced substance P-positive fibers). Moreover, S. boulardii CNCM I-745 diminished the production of HSV-1 associated pro-inflammatory cytokines in the myenteric plexus and increased levels of anti-inflammatory interleukins.ConclusionsS. boulardii CNCM I-745 ameliorated gastrointestinal neuromuscular anomalies in a mouse model of gut dysfunctions typically observed with irritable bowel syndrome.

Mitochondrial remodeling in the liver following chronic alcohol feeding to rats

The feeding of alcohol orally (Lieber-DeCarli diet) to rats has been shown to cause declines in mitochondrial respiration (state III), decreased expression of respiratory complexes, and decreased respiratory control ratios (RCR) in liver mitochondria. These declines and other mitochondrial alterations have led to the hypothesis that alcohol feeding causes “mitochondrial dysfunction” in the liver. If oral alcohol feeding leads to mitochondrial dysfunction, one would predict that increasing alcohol delivery by intragastric (IG) alcohol feeding to rats would cause greater declines in mitochondrial bioenergetics in the liver. In this study, we examined the mitochondrial alterations that occur in rats fed alcohol both orally and intragastrically. Oral alcohol feeding decreased glutamate/malate-, acetaldehyde- and succinate-driven state III respiration, RCR, and expression of respiratory complexes (I, III, IV, V) in liver mitochondria, in agreement with previous results. IG alcohol feeding, on the other hand, caused a slight increase in glutamate/malate-driven respiration, and significantly increased acetaldehyde-driven respiration in liver mitochondria. IG feeding also caused liver mitochondria to experience a decline in succinate-driven respiration, but these decreases were smaller than those observed with oral alcohol feeding. Surprisingly, oral and IG alcohol feeding to rats increased mitochondrial respiration using other substrates, including glycerol-3-phosphate (which delivers electrons from cytoplasmic NADH to mitochondria) and octanoate (a substrate for beta-oxidation). The enhancement of glycerol-3-phosphate- and octanoate-driven respiration suggests that liver mitochondria remodeled in response to alcohol feeding. In support of this notion, we observed that IG alcohol feeding also increased expression of mitochondrial glycerol phosphate dehydrogenase-2 (GPD2), transcription factor A (TFAM), and increased mitochondrial NAD+-NADH and NADP+-NADPH levels in the liver. Our findings suggest that mitochondrial dysfunction represents an incomplete picture of mitochondrial dynamics that occur in the liver following alcohol feeding. While alcohol feeding causes some mitochondrial dysfunction (i.e. succinate-driven respiration), our work suggests that the major consequence of alcohol feeding is mitochondrial remodeling in the liver as an adaptation. This mitochondrial remodeling may play an important role in the enhanced alcohol metabolism and other adaptations in the liver that develop with alcohol intake.

MCH receptor deletion does not impair glucose-conditioned flavor preferences in mice

The post-oral actions of glucose stimulate intake and condition flavor preferences in rodents. Hypothalamic melanin-concentrating hormone (MCH) neurons are implicated in sugar reward, and this study investigated their involvement in glucose preference conditioning in mice. In Exp. 1 MCH receptor 1 knockout (KO) and C57BL/6 wildtype (WT) mice learned to prefer 8% glucose over an initially more-preferred non-nutritive 0.1% sucralose+saccharin (S+S) solution. In contrast, the KO and WT mice preferred S+S to 8% fructose, which is consistent with this sugar's weak post-oral reinforcing action. In Exp. 2 KO and WT mice were trained to drink a flavored solution (CS+) paired with intragastric (IG) infusion of 16% glucose and a different flavored solution (CS−) paired with IG water. Both groups drank more CS+ than CS− in training and preferred the CS+ to CS− in a 2-bottle test. These results indicate that MCH receptor signaling is not required for flavor preferences conditioned by the post-oral actions of glucose. This contrasts with other findings implicating MCH signaling in other types of sugar reward processing.

Intragastric injection of Lactobacillus casei strain Shirota suppressed spleen sympathetic activation by central corticotrophin-releasing factor or peripheral 2-deoxy-d-glucose in anesthetized rats

Intragastric (IG) administration of probiotic strain Lactobacillus casei Shirota (LcS) decreases the sympathetic nerve outflow of anesthetized rats in a tissue-specific manner. In the present study, we examined the effects of IG administration of LcS on sympathetic activation induced by an intracerebroventricular (ICV) injection of corticotrophin-releasing factor (CRF) and an intravenous (IV) injection of 2-deoxy-d-glucose (2DG) or interleukin (IL)-1β in urethane-anesthetized rats. The IG administration of LcS differently affected the stimulatory responses of sympathetic nerve outflow to CRF. LcS suppressed the increase in splenic sympathetic nerve activity (Spleen-SNA), induced by central CRF, in a dose-dependent manner; however, it did not alter adrenal sympathetic nervous activity (ASNA). In contrast, LcS did not affect spleen-SNA and ASNA following an IV injection of IL-1β. On the other hand, IG administration of LcS suppressed the activation of ASNA following an IV injection of 2DG. These findings suggest that the suppression of central CRF-induced sympathetic activation by LcS is tissue-specific. Moreover, it can suppress the 2DG-induced sympathetic activation. Furthermore, we found that stomach-specific vagotomy attenuates the suppressive effect of LcS on CRF-mediated spleen-SNA activation. Thus, the present study suggests that LcS administered to the stomach may act on the afferent vagal nerve and send afferent signals to the brain to regulate efferent SNA induced by sympathetic stimulators.

SGLT1 sugar transporter/sensor is required for post-oral glucose appetition.

Recent findings suggest that the intestinal sodium-glucose transporter 1 (SGLT1) glucose transporter and sensor mediates, in part, the appetite-stimulation actions of intragastric (IG) glucose and nonmetabolizable α-methyl-d-glucopyranoside (MDG) infusions in mice. Here, we investigated the role of SGLT1 in sugar conditioning using SGLT1 knockout (KO) and C57BL/6J wild-type (WT) mice. An initial experiment revealed that both KO and WT mice maintained on a very low-carbohydrate diet display normal preferences for saccharin, which was used in the flavored conditioned stimulus (CS) solutions. In experiment 2, mice were trained to drink one flavored solution (CS+) paired with an IG MDG infusion and a different flavored solution (CS-) paired with IG water infusion. In contrast to WT mice, KO mice decreased rather than increased the intake of the CS+ during training and failed to prefer the CS+ over the CS- in a choice test. In experiment 3, the KO mice also decreased their intake of a CS+ paired with IG glucose and avoided the CS+ in a choice test, unlike WT mice, which preferred the CS+ to CS-. In experiment 4, KO mice, like WT mice preferred a glucose + saccharin solution to a saccharin solution. These findings support the involvement of SGLT1 in post-oral glucose and MDG conditioning. The results also indicate that sugar malabsorption in KO mice has inhibitory effects on sugar intake but does not block their natural preference for sweet taste.

Operant licking for intragastric sugar infusions: Differential reinforcing actions of glucose, sucrose and fructose in mice

Intragastric (IG) flavor conditioning studies in rodents indicate that isocaloric sugar infusions differ in their reinforcing actions, with glucose and sucrose more potent than fructose. Here we determined if the sugars also differ in their ability to maintain operant self-administration by licking an empty spout for IG infusions. Food-restricted C57BL/6J mice were trained 1h/day to lick a food-baited spout, which triggered IG infusions of 16% sucrose. In testing, the mice licked an empty spout, which triggered IG infusions of different sugars. Mice shifted from sucrose to 16% glucose increased dry licking, whereas mice shifted to 16% fructose rapidly reduced licking to low levels. Other mice shifted from sucrose to IG water reduced licking more slowly but reached the same low levels. Thus IG fructose, like water, is not reinforcing to hungry mice. The more rapid decline in licking induced by fructose may be due to the sugar's satiating effects. Further tests revealed that the Glucose mice increased their dry licking when shifted from 16% to 8% glucose, and reduced their dry licking when shifted to 32% glucose. This may reflect caloric regulation and/or differences in satiation. The Glucose mice did not maintain caloric intake when tested with different sugars. They self-infused less sugar when shifted from 16% glucose to 16% sucrose, and even more so when shifted to 16% fructose. Reduced sucrose self-administration may occur because the fructose component of the disaccharide reduces its reinforcing potency. FVB mice also reduced operant licking when tested with 16% fructose, yet learned to prefer a flavor paired with IG fructose. These data indicate that sugars differ substantially in their ability to support IG self-administration and flavor preference learning. The same post-oral reinforcement process appears to mediate operant licking and flavor learning, although flavor learning provides a more sensitive measure of sugar reinforcement.

Sugar-induced cephalic-phase insulin release is mediated by a T1r2+T1r3-independent taste transduction pathway in mice.

Sensory stimulation from foods elicits cephalic phase responses, which facilitate digestion and nutrient assimilation. One such response, cephalic-phase insulin release (CPIR), enhances glucose tolerance. Little is known about the chemosensory mechanisms that activate CPIR. We studied the contribution of the sweet taste receptor (T1r2+T1r3) to sugar-induced CPIR in C57BL/6 (B6) and T1r3 knockout (KO) mice. First, we measured insulin release and glucose tolerance following oral (i.e., normal ingestion) or intragastric (IG) administration of 2.8 M glucose. Both groups of mice exhibited a CPIR following oral but not IG administration, and this CPIR improved glucose tolerance. Second, we examined the specificity of CPIR. Both mouse groups exhibited a CPIR following oral administration of 1 M glucose and 1 M sucrose but not 1 M fructose or water alone. Third, we studied behavioral attraction to the same three sugar solutions in short-term acceptability tests. B6 mice licked more avidly for the sugar solutions than for water, whereas T1r3 KO mice licked no more for the sugar solutions than for water. Finally, we examined chorda tympani (CT) nerve responses to each of the sugars. Both mouse groups exhibited CT nerve responses to the sugars, although those of B6 mice were stronger. We propose that mice possess two taste transduction pathways for sugars. One mediates behavioral attraction to sugars and requires an intact T1r2+T1r3. The other mediates CPIR but does not require an intact T1r2+T1r3. If the latter taste transduction pathway exists in humans, it should provide opportunities for the development of new treatments for controlling blood sugar.

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 5ml 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.

Ghrelin signaling is not essential for sugar or fat conditioned flavor preferences in mice

The oral and post-oral actions of sugar and fat stimulate intake and condition flavor preferences in rodents through a process referred to as appetition. Ghrelin is implicated in food reward processing, and this study investigated its involvement in nutrient conditioning in mice. In Exp. 1 ghrelin receptor-null (GHSR-null) and C57BL/6 wildtype (WT) mice learned to prefer a flavor (CS+) mixed into 8% glucose over another flavor (CS−) mixed into a “sweeter” but non-nutritive 0.1% sucralose+saccharin (S+S) solution. In Exp. 2 treating WT mice with a ghrelin receptor antagonist [(D-Lys3)-GHRP-6] during flavor training did not prevent them from learning to prefer the CS+ glucose over the CS−S+S flavor. GHSR-null and WT mice were trained in Exp. 3 to drink a CS+ paired with intragastric (IG) infusion of 16% glucose and a CS− paired with IG water. Both groups drank more CS+ than CS− in training and preferred the CS+ to CS− in a choice test. The same (Exp. 4) and new (Exp. 5) GHSR-null and WT mice learned to prefer a CS+ flavor paired with IG fat (Intralipid) over a CS− flavor paired with IG water. GHSR-null and WT mice also learned to prefer a CS+ flavor added to 8% fructose over a CS− added to water. Together, these results indicate that ghrelin receptor signaling is not required for flavor preferences conditioned by the oral or post-oral actions of sugar and fat. This contrasts with other findings implicating ghrelin signaling in food reward processing and food-conditioned place preferences.

Intragastric fat self-administration is impaired in GPR40/120 double knockout mice

Mice acquire strong preferences for flavors paired with intragastric (IG) fat infusions. This IG fat conditioning is attenuated in double knockout (DoKO) mice missing GPR40 and GPR120 fatty acid receptors. Here we determined if GPR40/120 DoKO mice are also impaired in IG fat self-administration in an operant lick task. In daily 1-h sessions the mice were trained with a sipper spout that contained dry food pellets; licks on the spout triggered infusions of IG fat (Intralipid). The training sessions were followed by test sessions with an empty spout. GPR40/120 DoKO mice self-infused more 20% fat than wild type (WT) C57BL/6 mice in training with a food-baited spout (2.4 vs. 2.0kcal/h) but self-infused less 20% fat than WT mice in empty spout tests (1.2 vs. 1.7kcal/h). The DoKO mice also self-infused less 5% fat than WT mice (0.6 vs. 1.3kcal/h) although both groups emitted more licks for 5% fat than 20% fat. The DoKO and WT mice did not differ, however, in their self-infusion of 12.5% glucose (1.5 vs. 1.6kcal/h), which is isocaloric to 5% fat. A second 5% IL test showed that the DoKO mice reverted to a reduced self-infusion compared to WT mice. When the infusion was shifted to water, WT mice reduced licking in the first extinction session, whereas DoKO mice were less sensitive to the absence of infused fat. Our results indicate that post-oral GPR40/120 signaling is not required to process IG fat infusions in food-baited spout training sessions but contributes to post-oral fat reinforcement in empty spout tests and flavor conditioning tests.

Stem cell intervention ameliorates epigallocatechin-3-gallate/lipopolysaccharide-induced hepatotoxicity in mice.

Stem cells are identified as a novel cell therapy for regenerative medicine because of their ability to differentiate into many functional cell types. We have shown earlier a new model of hepatotoxicity in mice by administering (1500 mg/kg) epigallocatechin-3-gallate (EGCG) intragastric (IG) for 5 days after a single intraperitoneal dose (6 mg/kg) of lipopolysaccharide (LPS). In this study, we aimed to study the effect of intrahepatic (IH) injection of mouse embryonic stem cells (MESCs) on the hepatotoxicity induced by EGCG/LPS in mice. Mice were administered EGCG/LPS and rested for 3 days. MESCs were obtained from American Type Culture Collection and cultured in vitro for 4 days. Stem cells were injected IH. Seven days later, a single dose of LPS (6 mg/kg) followed by daily doses of IG administration of EGCG were re-administered for 5 days. At the end of the experiment, blood samples were collected for analysis of biochemical parameters associated with liver. Results showed that the group of mice that were administered MESCs prior to EGCG/LPS showed lower levels of alanine amino transferase, alkaline phosphatase, and bilirubin, higher albumin/globulin ratio, and less remarkable histopathological lesions. Also, that group of mice showed less expression of oxidative stress biomarkers (oxidized low-density lipoprotein Ox.LDL and chemokine CXCL16), less expression of nuclear protein receptors (retinoic acid receptor and retinoid X receptor), and less expression of inflammatory biomarkers (tumor necrosis factor α and transforming growth factor β1) compared with other groups of mice that were not given MESCs. In conclusion, MESCs can ameliorate EGCG/LPS-induced hepatotoxicity in mice.

Flavor Preferences in Animals: Role of Mouth and Gut Nutrient Sensors

Flavor Preferences in Animals: Role of Mouth and Gut Nutrient Sensors

Once a daily Tablet Formulation and In Vitro Evaluation of HPMC Based Intra Gastric Floating Tablet of Levofloxacin

The aim of this study was to develop a new intra-gastric floating tablet for controlled delivery of Levofloxacin for the treatment of peptic ulcer disease caused by Helicobacter pylori (H. pylori). The method of preparation is direct compression method. HPMC, K-grade and effervescent material sodium bicarbonate formed the floating layer. The release layer contained Levofloxacin and various polymers such as HPMC-K15M, HPMC-K100M, PVP-K30 and MCC in combination with the drug. The in vitro drug release was studied in pH 1.2 HCl using USP dissolution Apparatus II at 50 rpm. Zero-order, first-order, Higuchi and Korsmeyer et al. models were used to estimate the kinetics of drug release. Optimized formulation released approximately 98% drug in 12 h in vitro, while the floating lag time was 49 sec and the tablet remained floatable throughout all studies. Optimized formulation (D3) followed the Korsmeyer and Peppas model and showed no significant change in physical appearance, drug content, floatability and invitro dissolution pattern after storage at 45°C/75% RH for three month.

Flavor change and food deprivation are not critical for post-oral glucose appetition in mice

When mice trained to consume a CS− flavored solution paired with intragastric (IG) water self-infusion are given a new CS+ flavor paired with IG glucose self-infusion, their intake is stimulated within minutes in the first CS+ test. They also display a preference for the CS+ over the CS− in two-bottle tests. These indicators of post-oral appetite stimulation (appetition) have been studied in food-restricted mice, with novel CS+ and CS− flavors. Two experiments tested whether deprivation and flavor novelty are needed for stimulation of intake. Exp. 1 compared food-restricted and ad libitum fed C57BL/6 mice trained for 1h/day: 3 sessions with CS− flavor and IG water followed by 3 sessions with a novel CS+ flavor and IG 16% glucose. Ad libitum (AL) fed mice licked less overall, but like the food-restricted (FR) group they increased licking in the first session. In the choice test, FR mice displayed a significant CS+ preference (73%) whereas AL mice had a weaker preference (64%). In Exp. 2, food-restricted mice were trained with a flavor and IG water, and then the Same or a New flavor paired with IG 8% glucose. The glucose infusion rapidly stimulated intakes in the first and subsequent sessions and to the same degree in the two groups. Both groups also showed similar reductions in licking in extinction tests with IG water infusions. These data show that mice need not be explicitly food deprived or given a novel flavor cue to increase ongoing ingestion in response to post-oral glucose stimulation.

Pharmacokinetics of metronidazole in foals: influence of age within the neonatal period.

Neonatal foals have unique pharmacokinetics, which may lead to accumulation of certain drugs when adult horse dosage regimens are used. Given its lipophilic nature and requirement for hepatic metabolism, metronidazole may be one of these drugs. The purpose of this study was to determine the pharmacokinetic profiles of metronidazole in twelve healthy foals at 1-2.5 days of age when administered as a single intravenous (IV) and intragastric (IG) dose of 15 mg/kg. Foals in the intravenous group were studied a second time at 10-12 days of age to evaluate the influence of age on pharmacokinetics within the neonatal period. Blood samples were collected at serial time points after metronidazole administration. Metronidazole concentration in plasma was measured using LC-MS. Pharmacokinetic parameters were determined using noncompartmental analysis and compared between age groups. At 1-2.5 days of age, the mean peak plasma concentration after IV infusion was 18.79 ± 1.46 μg/mL, elimination half-life was 11.8 ± 1.77 h, clearance was 0.84 ± 0.13 mL/min/kg and the volume of distribution (steady-state) was 0.87 ± 0.07 L/kg. At 10-12 days of age, the mean peak plasma concentration after IV infusion was 18.17 ± 1.42 μg/mL, elimination half-life was 9.07 ± 2.84 h, clearance was 1.14 ± 0.21 mL/min/kg and the volume of distribution (steady-state) was 0.88 ± 0.06 L/kg. Oral approximated bioavailability was 100%. Cmax and Tmax after oral dosing were 14.85 ± 0.54 μg/mL and 1.75 (1-4) h, respectively. The elimination half-life was longer and clearance was reduced in neonatal foals at 1-2.5 days as compared to 10-12 days of age (P = 0.006, P = 0.001, respectively). This study warrants consideration for altered dosing recommendations in foals, especially a longer interval (12 h).

Roles of NMDA and dopamine D1 and D2 receptors in the acquisition and expression of flavor preferences conditioned by oral glucose in rats

Animals learn to prefer flavors associated with the intake of sugar (sucrose, fructose, glucose) and fat (corn oil: CO) solutions. Conditioned flavor preferences (CFP) have been elicited for sugars based on orosensory (flavor–flavor: e.g., fructose-CFP) and post-ingestive (flavor–nutrient: e.g., intragastric (IG) glucose-CFP) processes. Dopamine (DA) D1, DA D2 and NMDA receptor antagonism differentially eliminate the acquisition and expression of fructose-CFP and IG glucose-CFP. However, pharmacological analysis of fat (CO)-CFP, mediated by both flavor–flavor and flavor–nutrient processes, indicated that acquisition and expression of fat-CFP were minimally affected by systemic DA D1 and D2 antagonists, and were reduced by NMDA antagonism. Therefore, the present study examined whether systemic DA D1 (SCH23390), DA D2 (raclopride) or NMDA (MK-801) receptor antagonists altered acquisition and/or expression of CFP induced by oral glucose that should be mediated by both flavor–flavor and flavor–nutrient processes. Oral glucose-CFP was elicited following by training rats to drink one novel flavor (CS+, e.g., cherry) mixed in 8% glucose and another flavor (CS−, e.g., grape) mixed in 2% glucose. In expression studies, food-restricted rats drank these solutions in one-bottle sessions (2h) over 10days. Subsequent two-bottle tests with the CS+ and CS− flavors mixed in 2% glucose occurred 0.5h after systemic administration of vehicle (VEH), SCH23390 (50–800nmol/kg), raclopride (50–800nmol/kg) or MK-801 (50–200μg/kg). Rats displayed a robust CS+ preference following VEH treatment (94–95%) which was significantly though marginally attenuated by SCH23390 (67–70%), raclopride (77%) or MK-801 (70%) at doses that also markedly reduced overall CS intake. In separate acquisition studies, rats received VEH, SCH23390 (50–400nmol/kg), raclopride (50–400nmol/kg) or MK-801 (100μg/kg) 0.5h prior to ten 1-bottle training trials with CS+/8%G and CS−/2%G training solutions that was followed by six 2-bottle CS+ vs. CS− tests in 2% glucose conducted without injections. The significant and persistent CS+ preferences observed in the VEH (94–98%) group was significantly reduced by rats receiving SCH23390 at 400nmol/kg (65–73%), raclopride at 200 or 400nmol/kg (76–82%) or MK-801 at 100μg/kg (68–69%). Thus, systemic DA D1 and DA D2 receptor antagonism produced smaller reductions in the expression of oral glucose-CFP relative to fructose-CFP or IG-glucose-CFP. Correspondingly, systemic DA D1, DA D2 and NMDA receptor antagonism also produced smaller reductions in the acquisition of oral glucose-CFP relative to fructose-CFP or IG-glucose-CFP. These data suggest, but do not prove, that the magnitude and persistence of these receptor antagonist effects upon sugar-CFP might depend upon the individual or combined engagement of flavor–flavor and flavor–nutrient processes.

Time-course of extracellular nicotine and cotinine levels in rat brain following administration of nicotine: effects of route and ethanol coadministration.

Nicotine and ethanol are commonly coabused drugs, and nicotine-laced ethanol products are growing in popularity. However, little is known about time-course changes in extracellular nicotine and cotinine levels in rat models of ethanol and nicotine coabuse. The objective of the present study was to determine the time-course changes in brain levels of nicotine and cotinine following subcutaneous (SC) and intragastric (IG) nicotine administration in alcohol-preferring (P) and Wistar rats. In vivo microdialysis was used to collect dialysate samples from the nucleus accumbens shell (NACsh) for nicotine and cotinine determinations, following SC administration of (-)-nicotine (0.18, 0.35, and 0.70mg/kg) in female P and Wistar rats or IG administration of (-)-nicotine (0.35 and 0.70mg/kg) in 15% (v/v) ethanol or water in female P rats. SC nicotine produced nicotine and cotinine dialysate levels as high as 51 and 14ng/ml, respectively. IG administration of 15% EtOH + 0.70mg/kg nicotine in P rats resulted in maximal nicotine and cotinine dialysate levels of 19 and 14ng/ml, respectively, whereas administration of 0.70mg/kg nicotine in water resulted in maximal nicotine and cotinine levels of 21 and 25ng/ml, respectively. Nicotine and cotinine levels were detectable within the first 15 and 45min, respectively, after IG administration. Overall, the results of this study suggest that nicotine is rapidly adsorbed and produces relevant extracellular brain concentrations of nicotine and its pharmacologically active metabolite, cotinine. The persisting high brain concentrations of cotinine may contribute to nicotine addiction.

Rapid post-oral stimulation of intake and flavor conditioning in rats by glucose but not a non-metabolizable glucose analog

Mice adapted to drink a flavored saccharin solution (CS−) paired with intragastric (IG) self-infusions of water rapidly increase their intake of a new flavored solution (CS+) that is paired with IG glucose self-infusions. The present study extends this method to examine post-oral glucose appetition in rats. Food-restricted rats were trained to consume a CS− flavor (e.g., grape saccharin) paired with IG water in 5 daily 1-h tests. In the next 3 tests, they drank a CS+ (e.g., cherry saccharin) paired with IG glucose. Rats infused with 8% glucose increased intake significantly on CS+ Test 1, but those infused with 16% glucose showed only a small increase in intake, which may reflect a counteracting satiating effect. Both groups further increased CS+ intakes in Tests 2 and 3, and preferred (81%) the CS+ to the CS− in a two-bottle test without infusions. A second experiment investigated rats' responses to IG alpha-methyl-d-glucopyranoside (MDG), a non-metabolizable sugar analog which stimulates CS+ intake and preference in mice. The rats reduced their intake of the MDG-paired CS+ flavor over sessions, and preferred the CS− to the CS+ in the choice test. The glucose data show that rats, like mice, rapidly detect the sugar's positive post-oral effects that can stimulate intake within the first hour of exposure. The MDG avoidance may indicate a greater sensitivity to its post-oral inhibitory effects in rats than in mice, or perhaps slower clearance of MDG in rats. The test protocol described here can be used to investigate the peripheral and central processes involved in stimulation of intake by post-oral nutrients in rats.