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Poster Board No. 51 - POPULATION GENETICS AND MIGRATION OF PEROMYCUS LEUCOPUS, A LYME DISEASE RESERVOIR SPECIES. Keaka R. Farleigh, kfarleigh@capital.edu, (Christine S. Anderson, canders2@capital.edu), Capital University, Department of Biological [...]

It has been suggested that high-protein (HP) diets may favor weight management by lowering energy intake and reducing body fat. Whether these effects result from changes in energy metabolism remains unclear. We measured the adaptation of energy metabolism components during 2 wk of HP feeding. Fifty male Wistar rats were switched from a control diet to an HP diet (14 and 55% of protein, respectively) for 1,3, 6, or 14 d. Energy expenditure (EE) and substrate oxidation were measured by indirect calorimetry in feed-deprived rats and after consumption of a test meal. EE components, including the thermic effect of feeding and activity, were not modified during adaptation to an HP diet. Nutrient oxidation in feed-deprived rats was not affected by HP feeding, except for an early increase in protein oxidation. After 1 d, the postprandial inhibition of lipid oxidation (Lox) was blunted, carbohydrate (CHO) oxidation decreased by one-half, and urea clearance decreased by 66%. Thereafter, CHO oxidation gradually rose, resulting in a null CHO balance. Lox and urea clearance recovered after 3 d of adaptation to an HP diet, while protein oxidation reached a plateau. The postprandial oxidation of CliO counterbalanced the amount of ingested CHO as soon as 3 d, leading to a null postprandial CHO balance. We conclude that the inhibition of de novo lipogenesis from dietary CHO, but not EE and Lox, may participate in limiting the adiposity induced by HP feeding. The transient changes occurring during the period of adaptation to the diet highlight that the duration of the diet is critical in HP diet studies. doi: 10.3945/jn.109.120139.

Problem statement: In our earlier study, dietary pectin has been shown to promote nephrocalcinosis in rats when compared with the feeding of cellulose. It was not known whether this pectin effect relates to its structure or to its monosaccharide component, galacturonic acid. Approach: The effects of dietary pectin, galacturonic acid and glucose on kidney calcification were studied in female rats. The purified diets used had similar concentrations of calcium, magnesium and phosphorus. Results: It was confirmed that the feeding of pectin (10%, w/w) versus cellulose in diets containing 0.4% phosphorus results in significantly greater calcium and phosphorus concentrations in kidney. The new finding was that pectin had a nephrocalcinogenic effect when compared with either galacturonic acid or glucose. Conclusion: The stimulatory effect of pectin on the development of nephrocalcinosis is specific and is related to its structure rather than to its galacturonic acid component. Key words: Rats, nephrocalcinosis, diet, pectin, galacturonic acid, INTRODUCTION We have shown that the type of fiber influences nephrocalcinogenesis in female rats: Substitution of pectin for cellulose in a diet containing 0.4% phosphorus increased the severity of nephrocalcinosis [...]

In rats and humans, females have higher liver and/or plasma docosahexaenoic acid (DHA) content than males. We hypothesized that the effect of variation in total fat or essential fatty acid intakes on liver and plasma fatty acid composition would differ between sexes. Rats were fed a low-fat soybean oil (LFS), high-fat soybean oil (HFS), or high-fat linseed oil (HFL) diet for 20 d. There were significant sex differences in LFS rats in proportions of (n-3) and (n-6) fatty acids in plasma and liver contingent on lipid class. Significant diet x sex interactions were observed for eicosapentaenoic acid (EPA), DHA, and arachidonic acid (AA) status. HFL females had a higher proportion of EPA in plasma and liver phosphatidylcholine (PC), DHA in liver triacylglycerol (TAG), and AA in plasma PC than HFS and LFS females. These findings show that the effect of varying dietary fat intake on (n-3) and (n-6) long-chain PUFA (LCPUFA) status is modified by sex. Liver phospholipid and TAG fatty acid product:substrate ratios suggested greater [DELTA]6 desaturase ([DELTA]6D) activity in females than in males. The HFL diet induced higher [DELTA]6D mRNA expression compared with the LFS or HFS diets and HFL females had 10% higher expression of [DELTA]6D mRNA than HFL males. Together, these findings show that sex is an important determinant of the effect of variations in fat and fatty acid intake on LCPUFA status, which may have implications for recommendations for fat and fatty acid intake in humans. J. Nutr. 140: 245-250, 2010. doi: 10.3945/jn.109.115691.

This study comprised 2 experiments that tested the hypothesis that a high-amylose starch diet (AMO) would improve body weight and glycemic control relative to a high-amylopectin starch diet (AMN) in rats with diet-induced obesity. After inducing obesity with a high-fat and -energy diet (Expt. 1), male Sprague-Dawley rats (n = 46) were divided into 4 groups and given free or restricted access to either an AMN or an AMO diet for 4 wk (Expt. 2). After 3 wk, rats from each group underwent an oral glucose tolerance test. At the end of the experiment, food-deprived rats were killed by decapitation and blood and tissues were collected for analyses. AMO led to lower total energy intake, weight gain, fat pad mass, and glycemic response but higher insulin sensitivity index than AMN, only when consumed ad libitum (AL) (P < 0.05). AMO led to higher glucagon-like peptide-1 and peptide YY responses and mRNA levels, independent of feeding paradigm (P < 0.01). The mRNA levels of key neuropeptide systems involved in the regulation of food intake were affected only by energy restriction. On the other hand, AMO resulted in higher expression of uncoupling protein-1 in the brown adipose tissue than AMN in rats that consumed food AL (P < 0.05). The effects of AMO appear to be mediated by its high resistant starch content rather than its glycemic index. We conclude that starches high in AMO can be effective in weight and glycemic control in obesity.

In this study, our aim was to determine whether orally administered ovine serum Ig improved growth performance, organ weights, and gut morphology in growing rats and whether the method of manufacture of ovine serum Ig affected its bioactivity. Ninety Sprague-Dawley male rats were used in a 21-d growth study and were fed a basal control diet (BD; no Ig) and 5 test diets: spray-dried porcine plasma (SDPP), freeze-dried ovine Ig (FDOI), 2 concentrations of spray-dried ovine Ig ([SDO.sub.100] and [SDOI.sub.150]), and inactivated ovine Ig (IOI). Diets were isocaloric and contained the same amount of the first limiting amino acids, methionine plus cysteine. The body weight gain:feed ratio was higher (P < 0.05) for the FDOI-fed rats than for the BD- and IOI-fed groups. FDOI rats had higher jejunum (P < 0.05) and colon weights (P < 0.05) at the end of the study than rats in the BD group. Compared with the [SDOI.sub.100]-fed group, the FDOI group supported higher (P < 0.05) duodenum and colon weights. For gut morphology, the FDOI and the BD and IOI groups differed (P < 0.05). The FDOI-fed rats had longer (P < 0.05) villi and greater villi surface areas in the duodenum, jejunum, and ileum than the rats fed [SDOI.sub.100]. An ovine Ig fraction selectively improved growth performance, organ weight, and gut morphology in growing rats. Compared with spray-drying, a freeze-drying procedure appears to preserve a higher degree of immunological activity.

Dietary selenium (Se) requirements in rats have been based largely upon glutathione peroxidase-1 (Gpx1) enzyme activity and Gpx1 mRNA levels can also be used to determine Se requirements. The identification of the complete selenoprotein proteome suggests that we might identify additional useful molecular biomarkers for assessment of Se status. To characterize Se regulation of the entire rat selenoproteome, weanling male rats were fed a Se-deficient diet (

Although calcium (Ca) supplementation increases bone density, the increase is small and the effect on bone strength and fracture risk is uncertain. To investigate if bone mass, morphology, and biomechanical properties are affected by deficient to copious dietary Ca concentrations, the long bones (tibia and femur) of growing female Sprague-Dawley rats (8/group) were assessed after 13 wk of consuming 1, 2, 3, 4, 5, 6, or 7 g Ca/kg of a modified AIN-93G diet. Dietary phosphorous (P) and vitamin D remained constant at recommended concentrations. The assessment included mineralization, density, biomechanical properties of breaking by a 3-point flexure test, and morphological properties by microcomputed topography scanning of trabecular bone of the proximal tibia metaphysis. Dietary treatment did not affect food intake, weight gain, renal and muscle Ca concentrations, and bone hydroxyproline. All bone parameters measured were significantly impaired by Ca deficiency in rats fed the diet containing 1 g Ca/kg. Modest impairments occurred with some parameters (bone density, biomechanical bending moment, modulus of elasticity, and stress) in rats fed 2 g Ca/kg, but all parameters stabilized between 2 and 3 g/kg diet, with no differences between 3 and 7 g/kg. The results suggest that a threshold response in bone Ca retention or bone mass at ~2.5 g Ca/kg diet is associated with similar threshold responses in bone breaking strength and related biomechanics as well as trabecular structural properties. There was no evidence of a relative P deficiency or of improved or impaired bone strength and structure as Ca intakes increased beyond those needed to maximize bone density.

Our study investigated the potential for important in vivo drug-nutrient transport interactions at the lactating mammary gland using the L-carnitine transporter substrates, cefepime and L-carnitine, as proof-of-concept. On d 4 (n = 6/treatment) and d 10 (n = 6/treatment) of lactation, rats were administered cefepime (250 mg/h) or saline by continuous i.v. infusion (4 h). Serum and milk L-carnitine and cefepime concentrations were quantified by HPLC-UV. In whole mammary gland, organic cation/carnitine transporter (OCTN)I, OCTN2, OCTN3, amino acid transporter [B.sup.0,+] ([ATB.sup.0,+]), and h-carnitine transporter 2 expression were determined by quantitative RT-PCR and by western blot and immunohistochemistry when possible. Cefepime caused a 56% decrease in milk L-carnitine concentrations on lactation d 4 (P = 0.0048) but did not affect milk L-camitine at lactation d 10 or serum L-carnitine concentrations at either time. The mean L-carnitine and cefepime milk:serum ratios (M/S) decreased from 9.1 [+ or -] 0.4 to 4.9 [+ or -] 0.6 (P < 0.0001) and 0.89 [+ or -] 0.3 to 0.12 [+ or -] 0.02 (P = 0.0473), respectively, between d 4 and d 10 of lactation. In both groups, OCTN2 (P < 0.0001), OCTN3 (P = 0.0039), and [ATB.sup.0,+] (P = 0.004) mRNA expression and OCTN2 protein (P < 0.0001) were higher in mammary glands at d 4 of lactation compared with d 10. Immunohistochemistry revealed OCTN1 and OCTN2 localization in the mammary alveolar epithelium and OCTN3 expression in the interstitial space and blood vessel endothelium. In conclusion, cefepime significantly decreased milk L-carnitine concentrations only at d 4 of lactation. Relative to d 10, enhanced expression of OCTN2 and AT[B.sup.0,+] in mammary glands at d 4 of lactation and higher M/S (L-carnitine and cefepime) suggests cefepime competes with L-carnitine for L-carnitine transporters expressed in the lactating mammary gland to adversely affect L-carnitine milk concentrations and these effects depend upon lactation stage.

Recently, it was shown that the exposure to the potent hop phytoestrogen 8-prenylnaringenin (8-PN) depends on intestinal bacterial activation of isoxanthohumol (IX), but this occurs in only one-third of tested individuals. As the butyrate-producing Eubacterium limosum can produce 8-PN from IX, a probiotic strategy was applied to investigate whether 8-PN production could be increased in low 8-PN producers, thus balancing phytoestrogen exposure. Using fecal samples from high (Hop +) and low (Hop -) 8-PN-producing individuals, a Hop + and Hop--dynamic intestinal model was developed. In parallel, Hop + and Hop--human microbiota-associated rats were developed, germ-free (GF) rats acting as negative controls. IX and then IX + E. limosum were administered in the intestinal model and to the rats, and changes in 8-PN production and exposure were assessed. After dosing IX, 80% was converted into 8-PN in the Hop + model and highest 8-PN production, plasma concentrations, and urinary and fecal excretion occurred in the Hop + rats. Administration of the bacterium triggered 8-PN production in the GF rats and increased 8-PN production in the Hop--model and Hop rats. 8-PN excretion was similar in the feces (294.1 [+ or -] 132.2 nmol/d) and urine (8.5 [+ or -] 1.1 nmol/d) of all rats (n = 18). In addition, butyrate production increased in all rats. In conclusion, intestinal microbiota determined 8-PN production and exposure after IX intake. Moreover, E. limosum administration increased 8-PN production in low producers, resulting in similar 8-PN production in all rats.

Recent studies describe an association between poor iron status and obesity in humans, although the mechanism explaining this relationship is unclear. The present study aimed to determine the effect of moderate iron deficiency and physical activity (PA) on body composition in an animal model. Male Sprague-Dawley rats consumed iron-adequate (IA; 40 mg/kg) or moderately iron-deficient (ID; 9 mg/kg) diets ad libitum for 12 wk. Rats were assigned to 4 treatment groups (n = 10 per group): IA, sedentary (IAS); IA, PA (IAPA); ID, sedentary (IDS); or ID, PA (IDPA). Activity involved running on motorized running wheels at 4 m/min for 1 h/d for 5 d/wk. After 12 wk, ID rats were not anemic, but body iron stores were reduced as indicated by diminished (P < 0.05) femur iron compared with IA rats. Treatment group did not affect body weight or feed consumption. However, fat mass was greater (P < 0.05) in IDS rats (38.6 [+ or -] 6.7%) than IAS (31.8 [+ or -] 2.9%), IAPA (31.8 [+ or -] 2.0%), and IDPA (32.8 [+ or -] 4.5%) rats. Furthermore, lean body mass was diminished in IDS rats (58.7 [+ or -] 6.8%) compared with IAS (65.6 [+ or -] 3.0%), IAPA (65.6 [+ or -] 2.1%), and IDPA (64.7 [+ or -] 4.5%) rats. Thus, moderate iron deficiency may cause increased body fat accretion in rats and PA attenuates that effect.

It has been reported previously that the offspring of rat dams consuming low dietary copper (Cu) during pregnancy and lactation experience a deficiency in cardiac cytochrome c oxidase (CCO) characterized by reduced catalytic activity and mitochondrial and nuclear subunit content after postnatal d 10. The present study was undertaken to determine whether the cardiac CCO deficiency was caused directly by low postnatal Cu intake or whether it was a prenatal effect of low Cu intake by the dams that became manifest postnatally. Dams were fed either a Cu-adequate diet (6 mg Cu/kg) or Cu- deficient diet (1 mg Cu/kg) beginning 3 wk before conception and throughout gestation and lactation. One day following parturition, several litters from Cu-adequate dams were cross fostered to Cu-deficient dams and several litters from Cu-deficient dams were cross fostered to Cu-adequate dams. Litters that remained with their birth dams served as controls. CCO activity, the content of the mitochondrial-encoded CCO subunit 1 (COX1), and the content of the nuclear- encoded subunit COX4 in cardiac mitochondria were reduced in the 21-d-old offspring of Cu-deficient dams. COX1 content was normal in the 21-d-old cross-fostered offspring of Cu-deficient dams, but CCO activity and COX4 were reduced. Cross fostering the offspring of Cu-adequate dams to Cu-deficient dams did not significantly affect CCO activity, COX1 content, or COX4 content in cardiac mitochondria of 21-d-old offspring. These data indicate that low prenatal Cu intake by dams was the determinant of CCO activity in cardiac mitochondria of the 21-d-old offspring and may have led to the assembly of a less-than-fully active holoenzyme.

Foods of plant origin contain a large number of phytochemicals that may positively affect health. Phytochemicals are largely excreted in urine as metabolites that are formed in host tissues or by the microbiota and constitute a great proportion of the urinary metabolome. The latter can be characterized by a metabolomics approach. In this work, we compared the metabolism of lignins to that of the structurally related ferulic acid (FA) and sinapic acid (SA). Five groups of rats (n = 5) were fed for 2 d a purified diet alone [control (C)] or supplemented with lignin-enriched wheat bran (3% of the diet, wt:wt), poplar wood lignins (0.42%), FA (0.42%), or SA (0.42%). The metabolomes of urine samples collected after 1 and 2 d of supplementation were analyzed by high-resolution MS (liquid chromatography/quadrupole time-of-flight). Comparing metabolic fingerprints by gathering semiquantitative information on several hundreds of metabolites and using multivariate statistical analysis (partial least squares for discriminant analysis) showed the similarity between both lignin-supplemented and C groups and confirmed that lignins are largely inert and not absorbed in the body. One the other hand, metabolic fingerprints of the 2 phenolic acid-supplemented groups were clearly distinct from the C group. Differences between the groups were mainly from nonmetabolized FA and SA and metabolites excreted in urine. Thirteen of them were identified as sulfate esters and glucuronide and glycine conjugates of the same phenolic acids, and of dihydrosinapic, vanillic, and benzoic acids. This study shows that metabolomics allows the identification of new metabolites of phytochemicals and can be used to distinguish individuals fed different phytochemicaFcontaining foods.

This study determined the contribution of chronic salt loading and early loss of ovarian hormones on diastolic function in the hypertensive female mRen(2). Lewis rat, a monogenetic strain that expresses the mouse renin-2 gene in various tissues. Estrogen-intact mRen2 rats fed a high salt (HS) (8% sodium chloride) diet exhibited early diastolic dysfunction when compared to normal salt-fed (NS) (1% sodium chloride) rats. In contrast, ovariectomized (OVX) rats on either NS or HS diets showed impaired relaxation with evidence of elevated left ventricular filling pressures (E/e') or pseudonormalization. This more advanced stage of diastolic dysfunction was associated with increases in interstitial cardiac fibrosis and high circulating levels of aldosterone, two factors leading to reduced ventricular compliance. These findings may explain the preponderance of diastolic dysfunction and diastolic heart failure in postmenopausal women and provide a potential animal model for evaluating prevention and treatment interventions for this disorder. Key Words: Salt--Hormone, ovarian--Diastolic function--Rat.

Oxidative processes deteriorate the quality of meat products. High tannin sorghums (HTS) contain flavonoid oligomers known as proanthocyanidins of condensed tannins. These compounds act as antioxidants in vitro, but their effectiveness in vivo remains unclear. We tested the hypothesis that moderate amounts of dietary HTS could reduce markers of oxidation on muscle of rats without having detrimental effects in growth. We used 2 groups of 38 male Sprague Dawley rats at 5 and 13 wk of age each. Each age group was fed 4 diets in a completely randomized design. The younger group was fed the experimental diets for 10 wk (10W); whereas the older group was fed for 2 wk (2W). The diets were modified from the NIH-07 diet and contained HTS and corn at ratios of 0:50 (SO, control), 20:30 (S20), 35:15 (S35), and 50:0 (S50) as a percentage of the diet. Growth and the efficiency of gain were assessed periodically measuring BW, ADFI, ADG, and G:F. Oxidation in muscle was measured in fresh tissue and after 6 d of aerobic-refrigerated storage. Muscles evaluated were LM and soleus (SM). Fresh liver was also evaluated. Thiobarbituric acid-reactive substances (TBARS) and carbonyl content were used as markers of lipid and protein oxidation, respectively. No differences in BW, ADFI, ADG, and G:F were observed in 2W rats. Greater (P < 0.05) ADFI and ADG were observed in 10W-S35 group between d 1 and 7 and greater BW (P = 0.049) was observed in group 10W-S35 at d 70 compared with 10W-S0. No differences were observed between SO and any HTS diet in G:F in 10W and 2W rats. No differences in TBARS or carbonyls were observed in liver. No differences in TBARS were observed in fresh and aged LM and SM. When LM samples were aged for 6 d, decreased carbonyl contents (P < 0.01) were observed in 10W-S35 and 10W-S50 diets compared with 10W-S0. Reductions in carbonyls were also observed in aged SM between 2W-S50 and 2W-S0 (P = 0.013). We concluded that inclusion of 35% HTS in the diet increased intake and growth rate of young, fast-growing rats without changing the efficiency of gain. Feeding HTS reduced markers of protein oxidation in rat muscle after 6 d of refrigerated storage. If similar results are observed in animals such as swine or cattle, the use of HTS as animal feed should be reassessed. Key words: growth, muscle, oxidation, rat, sorghum, tannin

CCK-1-receptor-deficient Otsuka Long-Evans Tokushima fatty (OLETF) rats are hyperphagic and exhibit a greater preference for sucrose compared with lean controls [Long-Evans Tokushima Otsuka (LETO)]. To directly assess motivation to work for sucrose reward in this model of obesity and type 2 diabetes, we examined the operant performance of OLETF rats at nondiabetic and prediabetic stages (14 and 24 wk of age, respectively) on fixed-ratio (FR) and progressive-ratio (PR) schedules of reinforcement. To evaluate the involvement of dopamine systems, the effects of the D1 receptor antagonist SCH23390 (100 and 200 nmol/kg ip) and the D2 receptor antagonist raclopride (200 and 400 nmol/kg ip), were also tested on PR responding for sucrose. Compared with age-matched LETO rats, 14-wk-old OLETF rats emitted more licks on the 'active' empty spout operant on the FR-10 schedule of reinforcement to obtain 0.01 M and 0.3 M sucrose and completed higher ratio requirements on the PR schedule to gain access to 0.3 M and 1.0 M sucrose. At 24 wk, this effect was limited to 1.0 M sucrose. Both antagonists were potent in reducing operant responding to 0.3 M sucrose in both strains at both ages, and there was no strain effect to SCH23390 at either age. OLETF rats, on the other hand, showed an increased sensitivity to the higher dose of raclopride, resulting in reduced responding to sucrose reinforcement at 24 wk. Taken together, these findings provide the first direct evidence for an increased motivation for sucrose reward in the OLETF rats and suggest altered D2 receptor regulation with the progression of obesity and prediabetes. dietary obesity; food reward; appetite; palatability; operant lick task

Chronic administration of anorexigenic substances to experimental animals by injections or continuous infusion typically produces no effect or a transient reduction in daily food intake and body weight. Our aim was to identify an intermittent dosing strategy for intraperitoneal infusion of salmon calcitonin (sCT), a homolog of amylin that produces a sustained 25-35% reduction in daily food intake and adiposity in diet-induced obese rats. Rats (649 [+ or -] 10 g body wt, 27 [+ or -] 1% body fat), with intraperitoneal catheters tethered to infusion swivels, had free access to a 45% fat diet. Food intake, body weight, and adiposity during the 7-wk test period were relatively stable in the vehicle-treated rats (n = 16). None of 10 sCT dosing regimens administered in succession to a second group of rats (n = 18) produced a sustained 25-35% reduction in daily food intake for >5 days, although body weight and adiposity were reduced by 9% (587 [+ or -] 12 vs. 651 [+ or -] 14 g) and 22% (20.6 [+ or -] 1.2 vs. 26.5 [+ or -] 1.1%), respectively, across the 7-wk period. The declining inhibitory effect of sCT on daily food intake with the 6-h interinfusion interval appeared to be due in part to an increase in food intake between infusions. The declining inhibitory effect of sCT on daily food intake with the 2- to 3-h interinfusion interval suggested possible receptor downregulation and tolerance to frequent sCT administration; however, food intake increased dramatically when sCT was discontinued for 1 day after apparent loss of treatment efficacy. Together, these results demonstrate the activation of a potent homeostatic response to increase food intake when sCT reduces food intake and energy reserves in diet-induced obese rats. amylin; anorexia; body weight; body fat

The effect of dietary peptides on gut endogenous nitrogen (N) flow (ENFL) and amino acid (AA) flow (EAAFL) was studied. Semisynthetic diets containing enzyme-hydrolyzed casein (HC; 11%) or a free AA mixture devoid of Asp and Ser (A1) or Gly and Ala (A2) were formulated to have similar AA compositions except for the excluded AA and similar dietary electrolyte balances ([Na.sup.+] + [K.sup.+] - [CI.sup.-]). A protein-free diet (PF) served as a control. Sprague-Dawley rats were given the diets 8 times/d for 10 min each hour for 7 d. Rats were killed and digesta were sampled (6 observations within each group) along the intestinal tract 6 h after the first meal on d 7. EAAFL and ENFL, estimated with reference to the dietary marker Ti[O.sub.2], were determined directly (PF, A1, and A2) or after centrifugation and ultrafiltration of the digesta (HC). Endogenous flows of Asp and Ser or Gly and Ala did not differ (P > 0.05) in any of the intestinal sections between rats fed PF and A1 or PF and A2, respectively, except in the stomach where Ser flow was greater for rats fed A1. Ileal endogenous flows for most of the AA and for N were higher (P < 0.05) for rats fed the HC diet compared with those for rats fed the PF, A1, or A2 diets, except for Phe, Tyr, Lys, which did not differ among the groups. Ileal EAAFL and ENFL were not influenced by body N balance per se but were affected by the presence in the gut of dietary peptides derived from casein.

The fatty acid composition of membrane phospholipids affects the physicochemical properties of the membrane and thus influences cell function. In this study, the effects of 1-4 sequential pregnancies on the phospholipid fatty acid compositions of the maternal liver and erythrocytes were determined in female rats fed diets containing [alpha]-linolenic acid (ALA), ALA and preformed docosahexaenoic acid (DHA; ALA+DHA), or minimal ALA (low ALA). Virgin females, fed the diets for commensurate durations, served as a control for reproduction. Liver and erythrocyte total phospholipid compositions were determined at weaning by TLC/GC. In both tissues, significant main effects of diet and reproductive status were detected for many fatty acids, but a significant interaction of diet, reproductive status, and duration of treatment (no. of reproductive cycles or equivalent time period for virgins) was detected only for DHA, 22:6(n-3). Primiparous dams fed the ALA and low ALA diet had decreased liver DHA content of 31% and 74%, respectively, compared with virgin females fed the ALA diet. Liver DHA did not decrease further after additional reproductive cycles. Liver DHA content was unchanged in parous dams fed the ALA+DHA diet, but virgin females fed this diet exhibited a 50% increase in liver DHA after 13 wk of treatment. Changes in erythrocyte DHA were of similar magnitude and time course to those observed in liver, suggesting that this tissue may serve as a marker for liver DHA status.

Adult-onset calorie restriction (A-CR) is an experimental model of life extension and healthy aging less explored, compared with calorie restriction begun at early ages, but one more realistic for human application. We examined the effect of A-CR on the aging rat kidney with respect to common structural age-dependent changes and the accumulation of mitochondrial enzyme abnormalities in tubular epithelial cells. A 40% calorie restriction was initiated in middle-aged rats, before the onset of significant age-related changes in the Fischer x Brown Norway rat kidney. This dietary intervention effectively reduced glomerulo-sclerosis and tubular atrophy within 6 mo and changed the rate of interstitial fibrosis formation within 1 yr and vascular wall thickening and the expression cytochrome c oxidase (COX)-deficient tubular epithelial cells in 18 mo compared with age-matched ad libitum-fed rats. Our histological approach (histochemical staining for mitochondrial enzyme activity and laser capture microdissection) coupled with mitochondrial DNA (mtDNA) PCR analyses demonstrated that COX-deficient renal tubular epithelial ceils accumulated mtDNA deletion mutations and that these cells contained unique, clonally expanded mtDNA deletion mutations. Renal tubular epithelial cells with mitochondrial abnormalities presented cellular characteristics indicative of physiological dysfunction. dietary restriction; cytochrome c oxidase; mtDNA deletion mutations doi:10.1152/ajprenal.00307.2006.

The trafficking of dietary fat was assessed in obesity-prone (OP) and obesity-resistant (OR) male and female rats. Test meals containing [1-[sup.14]C]palmitate were delivered through gastric feeding tubes while rats consumed a high-carbohydrate diet (HCD) or after 5 days of a high-fat diet (HFD). Over the subsequent 24 h, the appearance of [sup.14]C was followed in the GI tract, skeletal muscles (SM), liver, adipose tissues (AT), and expired C[O.sub.2]. There was no difference in the production of [sup.14]C[O.sub.2] between OP and OR rats consuming a HCD. However, after 5 days on HFD, OR rats produced significantly more [sup.14]C[O.sub.2] after the test meal than OP rats (P < 0.001 females, P = 0.03 males). The differential oxidation of dietary fat between OP and OR rats on HFD was not due to differences in absorption but rather was associated with preferential disposition of tracer to AT in OP rats. Measurements of lipoprotein lipase in part explained increased tracer uptake by AT in OP rats but were not consistent with increased SM tracer uptake in OR rats. Surprisingly, female rats oxidized more tracer than male rats irrespective of phenotype or diet. These results are consistent with the notion that differences in the partitioning of dietary fat between storage in AT and oxidation in SM and liver that develop shortly after the introduction of a HFD may in part underlie the differential tendency for OR and OP rats to gain weight on this diet. thinness; lipoprotein lipase; sex differences; dietary fat; high-fat diet

We fed 2 low protein diets (LPD) to rats during pregnancy and lactation, and compared food intake and reproductive performance in the dams, and long-term growth in their offspring. The L93 and LM76 LPDs were derived from the American Society of Nutrition's recommended AIN93G and a modified version of the AIN76A purified control diets, respectively. The LPDs contained 8% crude protein in the form of casein and differed in their fat and carbohydrate sources. The purified control diets contained 19% crude protein. A regular cereal-based diet was also included, therefore, a total of 5 groups were tested. Blood urea nitrogen concentrations in dams of both LPD groups were lower than their respective controls, confirming decreased protein intake. The LM76 diet lowered food consumption of dams and produced energy malnourishment during pregnancy that persisted throughout lactation. In contrast, the L93 diet produced energy malnourishment only during lactation. Offspring of both LPD groups exhibited lower birth weights than their respective controls. Despite initiating nutritional rehabilitation at weaning (d 28), perinatal administration of both low protein diets produced long-term reductions in the body weight of male offspring. Interestingly, in the female offspring, the LM76 diet reduced birth weight for the entire duration of the study (180 d), whereas the L93 diet produced a relatively short-term (up to 58 d) reduction in body weight. This suggests that the imprinting effect of the perinatal nutritional environment on body weight is diet and gender dependent. The performance of the purified control diet groups were similar to the nonpurified diet group in most measured biochemical indices, with the notable exception of a decrease in the body-weight normalized kidney weight of the dams.

Retinoic acid (RA), produced from vitamin A (VA, retinol), is required for normal lung development and postnatal lung maturation. The concentration of retinyl ester (RE), the major storage form of retinol, decreases in the lungs in the perinatal period. Previously, we tested VARA, a nutrient-metabolite combination of VA and RA (10:1 molar ratio), on lung RE formation in postnatal rats and showed that the components of VARA acted synergistically to increase lung RE, as compared with the effects of equal amounts of VA and RA given alone. In this study, we first determined the equivalency of orally administered VARA in comparison to a standard oral supplement of VA, with respect to lung and liver RE storage. In a dose-dilution study, VARA was 4 times as effective as the standard dose of VA (VARA-25% did not differ from VA100%). The synergistic effect of VARA was selective for the lungs, compared with the liver, in which VA and VARA had equal effects. Secondly, we tested whether the 2 components of VARA must be coadministered to exert their synergistic effect on lung RE content. RA and VA and were administered separately and together as VARA. Although RA alone had no effect on lung RE in this 24-h experiment, RA synergized with VA administered either 12 h before RA or 12 h after RA, as well as when coadministered as VARA. We infer that VA and RA are both limiting for lung RE formation in neonates. Given the importance of bioactive retinoids in cell differentiation and lung development, assuring an adequate lung RE content postnatally could be of benefit for lung maturation.

Almond preferences were produced by giving rats a mixture of almond and sucrose (Experiments 1-4) or saccharin (Experiment 4). A subsequent extinction procedure consisted of either repeated 2-bottle almond versus water tests (Experiment 1) or repeated exposure to almond alone (Experiments 2-4). The main independent variable was whether access to food following a session was given immediately, 30 min later, or 120 min later. No effect of extinction was found in any experiment. An important finding was that varying the delay until food access had no detectable effect. It was concluded that inadvertent flavor-food associations do not maintain preference for the flavor under extinction conditions. Keywords: flavor preference, extinction, delayed food, rats

Low tissue levels of (n-3) polyunsaturated fatty acids (PUFAs), particularly docosahexaenoic acid [DHA, 22:6(n-3)], are implicated in postpartum depression. The effects of 1-4 sequential reproductive cycles on maternal brain phospholipid fatty acid composition were determined in female rats fed diets containing [alpha]-linolenic acid (ALA), containing ALA and preformed DHA (ALA + DHA), or lacking ALA (low-ALA). Virgin females, fed the diets for commensurate durations served as a control for reproduction. Whole-brain total phospholipid composition was determined at weaning by TLC/GC. A single reproductive cycle on the low-ALA diet decreased brain DHA content by 18% compared to ALA primiparas (P < 0.05), accompanied by incorporation of docosapentaenoic acid ((n-6) DPA, 22:5(n-6)) to 280% of ALA primiparas (P < 0.05). DHA was not further decreased after subsequent cycles; however, there was an additional increase in (n-6) DPA after the second cycle (P < 0.05). Brain DHA of virgin females fed the low-ALA diet for 27 wk decreased 15% (P < 0.05), but was accompanied by a more modest increase in (n-6) DPA than in parous low-ALA dams (P < 0.05). Virgin females and parous dams fed the diet containing ALA + DHA exhibited only minor changes in brain fatty acid composition. These observations demonstrate that brain DHA content of adult animals is vulnerable to depletion under dietary conditions that supply inadequate (n-3) PUFAs, that this effect is augmented by the physiological demands of pregnancy and lactation, and that maternal diet and parity interact to affect maternal brain PUFA status.

Obesity is often associated with insulin resistance and mitochondrial dysfunction within skeletal muscles, but the causative factors are not clearly identified. The present study examined the role of nutrition, both qualitatively and quantitatively, in the induction of muscle mitochondrial defects. Two experimental diets [high sucrose (SU) and high fat (F)] were provided for 6 wk to male Wistar rats at 2 levels of energy [standard (N) and high (H)] and compared with a standard energy cornstarch-based diet (C). Insulin sensitivity (intraperitoneal glucose tolerance test, IPGTT) and intramyocellular triglyceride (IMTG) content ([sup.1]H MRS) were determined at wk 5. Mitochondrial oxidative phosphorylation and superoxide anion radical (MSR) production were assessed on soleus (oxidative) and tibialis (glycolytic) muscles. Experimental diets induced hyperinsulinemia during IPGTT (P < 0.01 vs. C). Rats in the HSU and HF groups were hyperglycemic relative to the C group, P < 0.05 vs. C. The severity of insulin resistance paralleled IMTG accumulation (P < 0.05). In soleus, mitochondrial respiration and ATP production rates were lower in HSU and HF than in C (P < 0.05). By contrast, respiration was unaffected by the diets in tibialis, whereas ATP production tended to be lower in rats fed the experimental diets compared with C (P = 0.09). Mitochondrial adaptations were associated with more than a 50% reduction in MSR production in HSU and HF compared with C in both soleus (P < 0.05) and tibialis (P < 0.01). Changes in mitochondrial functions in the NSU and NF groups were intermediate and not significantly different from C. Therefore, excess fat or sucrose and more importantly, excess energy intake by rats is associated with muscle type-specific mitochondrial adaptations, which contribute to decrease mitochondrial production of ATP and reactive oxygen species.

The effect of Bifidobacterium lactis (Bb-12) and Lactobacillus salivarius (UCC500) on the formation of carboxylic acids (CAs) was studied in the hindgut of rats fed pectin, inulin of low solubility, and lactitol. When the pectin diet was supplemented with Bb-12, the formation of CAs was larger throughout the colon of rats, due to increased formation of acetic acid (P < 0.01) and, in the distal part of the colon, also because of propionic and butyric acids (P < 0.01). In rats fed pectin and UCC500, there was a shift in the formation of CAs from the cecum to the distal colon. Thus, the cecal pool of CAs in the rats was lower (P < 0.05), whereas the concentration of CAs in the distal part of colon was larger (P < 0.01) than without this strain. Concerning the slowly fermentable inulin, there was a greater formation of CAs in the cecum (P < 0.05) of rats, especially propionic acid, and a lower formation in the distal part of the colon (P < 0.01) when the diets were supplemented with Bb-12, whereas UCC500 had no effect except for a lower proportion of acetic acid in the distal part of the colon (P < 0.001). In rats fed lactitol and Bb12, the concentration of CAs was lower in the distal part of colon (P < 0.001) than without this strain, whereas the cecal pool of CAs was greater in rats supplemented with UCC500 (P < 0.001). We conclude that the probiotic bacteria affect the amount, the pattern, and the site of release of CAs in the hindgut of rats, but the combination of pre- and probiotics is of great importance for the outcome.

We showed previously that [alpha]-eleostearic acid ([alpha]-ESA; 9Z11 E13 E-18:3) is converted to 9Z11 E-conjugated linoleic acid (CLA) in rats through a [DELTA] 13-saturation reaction. To investigate this further, we examined the absorption and metabolism of [alpha]-ESA in rat intestine using a lipid absorption assay in lymph from the thoracic duct. In this study, we used 4 test oils [tung oil, perilla oil, CLA-triacylglycerol (TG), and pomegranate seed oil, containing [alpha]-ESA, [alpha]-linolenic acid (LnA; 9Z12 Z15Z-18:3), CLA, and punicic acid (PA; 9Z11 E13Z-18:3), respectively]. Emulsions containing the test oils were administered to rats, and lymph from the thoracic duct was collected over 24 h. The positional and geometrical isomerism of CLA produced by PA metabolism was determined using GC-electron impact (EI)-MS and [sup.13]C-NMR, respectively; the product was confirmed to be 9Z11 E-CLA. A part of [alpha]-ESA and PA was converted to 9Z1 1E-CLA 1 h after administration; therefore the lymphatic recoveries of [alpha]-ESA and PA were modified by the amount of recovered CLA. Cumulative recovery of CLA, [alpha]-ESA, and PA was lower than that of LnA only during h 1 (P < 0.05), and cumulative recovery of [alpha]-ESA and PA was significantly lower than that of LnA and CLA for 8 h (P < 0.05). Therefore, the absorption rate was LnA > CLA > [alpha]-ESA = PA. The conversion ratio of [alpha]-ESA to 9Z11 E-CLA was higher than that of PA to 9Z11 E-CLA over 24 h (P < 0.05). These results indicated that [alpha]-ESA and PA are slowly absorbed in rat intestine, and a portion of these fatty acids is quickly converted to 9Z11 E-CLA.

The transsulfuration pathway, which aids in regulating homocysteine concentration and mediates cysteine synthesis, may be sensitive to vitamin B-6 status because cystathionine [beta]-synthase (CBS) and cystathionine [gamma]-lyase (CGL) require pyridoxal 5'-phosphate (PLP). To assess relations between vitamin B-6 and transsulfuration, we evaluated the effects of dietary pyridoxine (PN) on the hepatic concentration of relevant metabolites and in vitro activity of CBS and CGL. Growing rats were fed AIN-93G- or AIN-76A-based diets that ranged from adequate to deficient in vitamin B-6 (2, 1, 0.5, 0.1, or 0 mg of PN/kg diet, n = 5). This design allowed assessment of the effects of supplemental methionine (AIN-76A) vs. cysteine (AIN-93G) in common research diets over a range of vitamin B-6 levels. CBS activity, assayed in the presence or absence of added S-adenosylmethionine, was independent of diet type and PN level. CGL activity was independent of diet type but proportional to dietary PN. Rats fed deficient (0 and 0.1 mg PN/kg) diets exhibited only ~30% of the CGL activity of those fed the 2 mg PN/kg diets. Hepatic cystathionine increased from 20 to 30 nmol/g for the 1-2 mg PN/kg diets to ~85 nmol/g for the 0 mg PN/kg diet; however, cysteine was reduced only in B-6--deficient rats consuming the AIN-93G diet (means of 30-40 nmol/g for adequate to 11.6 nmol/g for 0 mg PN/kg AIN-76A diet). In spite of these effects, hepatic glutathione concentration increased in vitamin B-6 deficiency. These results suggest that vitamin B-6-dependent changes in transsulfuration do not limit hepatic glutathione production.

Although many animal studies have reported that dietary excess of methionine causes toxic changes including growth suppression and hemolytic anemia, the biochemical mechanism and biomarkers for methionine toxicity have not been well elucidated. The present study aimed to identify toxicity biomarkers from plasma metabolites in rats fed excessive methionine. Young growing rats were fed graded doses of additional methionine for 2 wk. Cluster analysis of multivariate correlations was performed on the physiological and toxicity variables with plasma metabolites detected by GC/MS, amino acid analyzer, and thiol-specific analysis. Indicative variables for hemolysis such as splenic nonheme iron content and plasma bilirubin were grouped in the same cluster as many methionine metabolites. Homocysteine and some undefined metabolites in this cluster were found to be strong discriminators between nontoxic and toxic levels of methionine intake. Product-to-precursor ratios of each methionine metabolite demonstrated that excessive methionine intake caused a marked decrease only in the ratio of cystathionine to homocysteine, suggesting that metabolism from homocysteine to cystathionine would be rate limiting in the disposal of excessive methionine. Collectively from these results, homocysteine appeared to be the most plausible biomarker to assess methionine excess as a surrogate marker both for toxicity and for setting a metabolic upper limit. KEY WORDS: * amino acid * dietary reference intake * tolerable upper level * acceptable daily intake * adequate intake

The health benefits of lycopene as an anticarcinogenic compound have been widely studied but little is known about the metabolic products of lycopene produced in vivo. We investigated lycopene metabolites in the liver of F344 male rats that had been prefed a lycopene-containing diet (0.25 g lycopene/kg diet). After 30 d of feeding, they were given a single oral dose of [sup.14]C-labeled lycopene (421.8 kBq). The metabolic products of both nonradioactive and [sup.14]C-labeled lycopene in rat liver were extracted and separated using HPLC and analyzed by UV/VIS spectrometry, online radioactive detection, and off-line and in-line positive ion electrospray ionization MS. Among a number of metabolite products formed, we identified apo-8'-lycopenal ([[lambda].sub.max] = 473 nm and m/z = 417). The putative compound, apo-12'-lycopenal, was detected but no apo-10'-lycopenal was present. A number of other very polar, short-chain and/ or short chromophore compounds with UV/VIS absorption KEY WORDS: * lycopene * lycopene metabolites * apo-8'-lycopenal * apo-12'-lycopenal * rats

Docosahexaenoic acid [22:6(n-3)] is enriched in brain membrane phospholipids and essential for brain function. Neurogenesis during embryonic and fetal development requires synthesis of large amounts of membrane phospholipid. We determined whether dietary (n-3) fatty acid deficiency during gestation alters neurogenesis in the embryonic rat brain. Female rats were fed diets with 1.3% energy [(n-3) control] or 0.02% energy [(n-3) deficient], from [alpha]-linolenic acid [18:3(n-3)], beginning 2 wk before gestation. Morphometric analyses were performed on embryonic day 19 to measure the mean thickness of the neuroepithelial proliferative zones corresponding to the cerebral cortex (ventricular and subventricular zones) and dentate gyrus (primary dentate neuroepithelium), and the thickness of the cortical plate and sectional area of the dentate gyrus. Phospholipids and fatty acids were determined by HPLC and GLC. Docosahexaenoic acid was 55-65% lower and (n-6) docosapentaenoic acid [22:5(n-6)] was 150-225% higher in brain phospholipids at embryonic day 19 in the (n-3) deficient (n 6 litters) than in the control (n = 5 litters) group. The mean thickness of the cortical plate and mean sectional area of the primordial dentate gyrus were 26 and 48% lower, respectively, and the mean thicknesses of the cortical ventricular zone and the primary dentate neuroepithelium were 110 and 70% higher, respectively, in the (n-3) deficient than in the control embryonic day 19 embryos. These studies demonstrate that (n-3) fatty acid deficiency alters neurogenesis in the embryonic rat brain, which could be explained by delay or inhibition of normal development. KEY WORDS: * (n-3) fatty acids * docosahexaenoic acid * neurogenesis * cerebral cortex * dentate gyrus

Aberrations in methylation profile of the genome occur in human cancers induced by folate deficiency. To elucidate the underlying mechanism, male F344 rats were fed a diet deficient in L-methionine and devoid of folic acid and choline (FMD diet), which is known to induce hepatocellular carcinomas. We investigated changes in the DNA methylation machinery, namely, de novo DNA methyltransferases (Dnmt3a and 3b), maintenance DNA methyltransferase (Dnmt1), and methyl CpG binding proteins (MBDs), in rat livers during early stages of tumorigenesis. RTPCR and Western blot analyses revealed differential expression of these proteins in the livers of rats fed the FMD diet. Although the hepatic Dnmt1 mRNA level declined with age (P < 0.001), it was elevated (P < 0.001) in deficient rats compared with controls. The changes in hepatic Dnmt1 protein level with the diet correlated with its mRNA levels (r = 0.60, P = 0.002). Similarly, the Dnmt3a mRNA level was elevated in rats fed the FMD diet (P < 0.001), whereas the Dnmt3b level (mRNA and protein) was not affected by diet or age. Compared with controls, hepatic MBD1-3 RNA levels increased (P < 0.001) and the protein levels of MBD1, 2, and 4 were elevated (P < 0.001) in the deficient rats. In both diet groups, hepatic MBD2 protein decreased (P < 0.001), whereas MeCP2 protein increased (P < 0.001) with age. These results demonstrate that a combined folate and methyl deficiency alters components of the DNA methylation machinery by both transcriptional and posttranscriptional mechanisms during early stages of hepatocarcinogenesis. KEY WORDS: * hepatocarcinogenesis * folate- and methyl-deficient diet * Dnmt1/3a/3b * MBD1-4

Foods high in resistant starch have the potential to improve human health and lower the risk of serious noninfectious diseases. RNA interference was used to down-regulate the two different isoforms of starch-branching enzyme (SBE) II (SBEIIa and SBEIIb) in wheat endosperm to raise its amylose content. Suppression of SBEIIb expression alone had no effect on amylose content; however, suppression of both SBEIIa and SBEIIb expression resulted in starch containing >70% amylose. When the >70% amylose wheat grain was fed to rats in a diet as a wholemeal, several indices of large-bowel function, including short-chain fatty acids, were improved relative to standard wholemeal wheat. These results indicate that this high-amylose wheat has a significant potential to improve human health through its resistant starch content. genetic engineering | nutrition | starch

Exposure to hypoxia induces anorexia in humans and rodents, but the role of leptin remains under discussion and that of orexigenic and anorexigenic hypothalamic neuropeptides remains unknown. The present study was designed to address this issue by using obese ([Lepr.sup.fa]/[Lepr.sup.fa]) Zucker rats, a rat model of genetic leptin receptor deficiency. Homozygous lean ([Lepr.sup.FA]/[Lepr.sup.FA]) and obese ([Lepr.sup.fa]/ [Lepr.sup.fa]) rats were randomly assigned to two groups, either kept at ambient pressure or exposed to hypobaric hypoxia for 1, 2, or 4 days (barometric pressure, 505 hPa). Food intake and body weight were recorded throughout the experiment. The expression of leptin and vascular endothelial growth factor (VEGF) genes was studied in adipose tissue with real-time quantitative PCR and that of selected orexigenic and anorexigenic neuropeptides was measured in the hypothalamus. Lean and obese rats exhibited a similar hypophagia (38 and 67% of initial values at day 1, respectively, P < 0.01) and initial decrease in body weight during hypoxia exposure. Hypoxia led to increased plasma leptin levels only in obese rats. This resulted from increased leptin gene expression in adipose tissue in response to hypoxia, in association with enhanced VEGF gene expression. Increased hypothalamic neuropeptide Y levels in lean rats 2 days after hypoxia exposure contributed to accounting for the enhanced food consumption. No significant changes occurred in the expression of other hypothalamic neuropeptides involved in the control of food intake. This study demonstrates unequivocally that altitude-induced anorexia cannot be ascribed to anorectic signals triggered by enhanced leptin production or alterations of hypothalamic neuropeptides involved in anabolic or catabolic pathways. altitude; anorexia; energy balance; adipose tissue

Garlic has been reported to have chemopreventive effects against a variety of cancers. However, different garlic preparations contain different constituents. We investigated the chemopreventive effect of aged garlic extract (AGE), an odorless product from prolonged extraction of fresh garlic, on colon carcinogenesis and cell proliferation in 1,2-dimethylhydrazine (DMH)-induced colon neoplastic rats. Rats were given weekly subcutaneous injections of DMH (20mg/kg) for 20 wk, and fed either a basal diet or one containing 4% AGE. Serum from AGE-treated rats contained detectable S-allylcysteine. The AGE diet significantly reduced the number of colon tumors and aberrant crypt foci compared to the basal diet. Cell proliferation of normal-appearing colonic mucosa was assessed by MIB-5 immunohistochemistry. AGE treatment significantly decreased the mean MIB-5-1abeling index. These findings suggest AGE has a chemopreventive effect on colon carcinogenesis through suppression of cell proliferation. KEY WORDS: * aged garlic extract * S-allylcysteine * colon carcinogenesis * chemoprevention * cell proliferation

A unique garlic preparation, aged garlic extract (AGE), was examined for its modifying effect on diethylnitrosamine (DEN)-induced neoplasia of the liver in male F344 rats, using the medium-term bioassay system based on the 2-step model of hepatocarcinogenesis. Carcinogenic potential was scored by comparing the numbers and areas of induced glutathione S-transferase placental form (GST-P)-positive hepatocellular foci. GST-P-positive foci were significantly decreased in rats treated with AGE at doses of 2, 5, and 10 mL/kg, i.g., 5 times per week during the promotion phase. In addition, to clarify the mechanism underlying the inhibitory effect of AGE, the effect of AGE on hepatocellular proliferation was evaluated using partially hepatectomized rats as a liver-regeneration model. The bromodeoxyuridine-labeling indices in the livers of the AGE group were significantly lower than those in the control group at 24 h, the maximum proliferation period after partial hepatectomy. These findings indicate that AGE inhibited the development of putative preneoplastic lesions in rat hepatocarcinogenesis, involving a slowing in the proliferation rate of liver cells after partial hepatectomy. KEY WORDS: * hepatocarcinogenesis * aged garlic extract * chemoprevention * medium-term bioassay system * glutathione S-transferase

We studied the effects of two garlic sources on systolic blood pressure (SBP) using spontaneously hypertensive rats (SHRs). Beginning at 12 wk of age, male SHRs were fed diets containing either aged garlic extract (AGE) or raw garlic (RG) powder for 10 wk. Both AGE and RG reduced the increase of SBP compared with the control group from 4 wk after beginning the experimental diets. The effect of AGE was accompanied by a decrease of pulse pressure (PP), suggesting an improvement of the pliability of the artery, although RG did not affect PP. However, harmful effects were observed in the RG group, including a decrease in erythrocytes, an increase in reticulocytes, and generation of papilloma in the forestomach. These results suggest that AGE may safely improve several factors related to blood vessel physiology and circulatory disease. KEY WORDS: * hypertension * aged garlic extract * rat

We investigated hypohomocysteinemic action as a cardiovascular protective property of aged garlic extract (AGE). Hyperhomocysteinemia was induced in rats by feeding folate-depleted diets. Plasma folate concentrations of 5, 24, and 202 nmol/L were detected in rats fed a folate-deficient L-amino acid diet containing succinyl sulfathiazole, an AIN-93G folate-deficient diet, and an AIN-93G folate-sufficient diet, respectively. Plasma concentrations of total homocysteine were elevated to the highest level (32 [micro]mol/L) by severe folate deficiency and to a moderate level (9 [micro]mol/L) by mild folate deficiency, compared with the lowest level of (5 [micro]mol/L), noted for the folate-sufficient group. The addition of AGE to the severely folate-deficient diet decreased plasma total homocysteine concentration by 30%. Hyperhomocysteinemia caused by mild folate deficiency remained unaltered by AGE supplementation. The reduction in total homocysteine of the severely folate-deficient rats was accompanied by a proportional decrease in protein-bound and free homocysteine, resulting in an unchanged protein-bound:free homocysteine ratio. AGE added to the diet did not alter plasma concentrations of other aminothiol compounds: cysteine, glutathione, and cysteinylglycine. These data, together with increased S-adenosylmethionine and decreased S-adenosylhomocysteine concentrations in the liver, suggest that the hypohomocysteinemic effect of AGE most likely stems from impaired remethylation of homocysteine to methionine and enhanced transsulfuration of homocysteine to cystathionine. More importantly, in addition to its cholesterol-lowering potential, blood pressure-lowering effect, and antioxidant property, a hypohomocysteinemic action may be another important cardiovascular protective factor of AGE. KEY WORDS: * folate deficiency * garlic * hyperhomocysteinemia * total homocysteine

Oligosaccharides are included among the anti-inflammatory components of milk because of their prebiotic properties and their capacity to act as receptors of microorganisms. Here the intestinal anti-inflammatory effect of goat milk oligosaccharides (O) was assessed in trinitrobenzenesulfonic (T) acid-induced colitis in rats. Rats were randomly assigned to three different groups. Two groups (T and OS) of colitic rats and a control group (C) were studied. Group OS received 500 mg/(kg*d) of goat milk oligosaccharides orally, starting 2 d before the colitis induction until d 6, and groups T and C received the vehicle. When compared with the T group, the OS group showed decreased anorexia and body weight loss; reduced bowel wall thickening and longitudinal extension of necrotic lesions; downregulated colonic expression of interleukin 1/3, inducible nitric oxide synthase, cyclooxygenase 2, and mucin 3; and increased trefoil factor 3. Thus, goat milk oligosaccharides have anti-inflammatory effects in rats with experimental colitis and may be useful in the management of inflammatory bowel disease. KEY WORDS: * colon *goat milk * inflammatory bowel disease *intestine * oligosaccharides

Selenium deficiency is associated with impaired bone metabolism and osteopenia in rats. However, it is not known how combined selenium and iodine deficiency affects bone metabolism. Therefore, we investigated the effect of selenium and iodine deficiency on bone metabolism in 2nd-generation selenium- and iodine-deficient rats. Selenium-deficient (Se-), iodine-deficient (I-), selenium- and iodine-deficient (Se-/I-), and control rats (Se+/I+), were pair-fed their respective diets until they were 74 d old. Each pair-fed rat was fed a selenium-adequate diet in the same amount as that consumed the day before by its selenium-deficient counterpart, taking food spillage into account. The skeletal phenotype was analyzed by dual energy X-ray absorptiometry, histomorphometry, and bone metabolism markers. Erythrocyte glutathione peroxidase activity (Gpx) and plasma thyroid hormones were measured to assess selenium and iodine status, respectively. In both Se-/I+ and Se-/I- rats, Gpx was reduced by 99% compared with pair-fed Se +/I+ and Se+/I- rats (P < 0.001). Iodine deficiency reduced plasma thyroxine by 64% in the 2 iodine-deficient groups (P < 0.001). Body weight, tail length, plasma insulin-like growth factor, pituitary growth hormone concentration, and femur and tibia bone mineral density were significantly greater in the Se-/I- rats than in the Se-/I+ rats. This study shows that iodine deficiency mitigated growth retardation and osteopenia in 2nd-generation selenium-deficient rats and suggests that adequate selenium status should be ensured before measures are taken to correct iodine deficiency. KEYWORDS: * seleniumdeficiency * iodinedeficiency * bonemineraldensity * osteopenia * bonemetabolism

The The mdx mouse is an animal model for Duchenne muscular dystrophy. Mdx mice fed a 12% NaCl diet from birth up to 20 days of age (mdx-Na mice) had an ~50% reduction in serum creatine kinase (CK) activity compared with mdx mice fed a standard diet. Most notably, necrotic fibers in tibialis anterior (TA) muscle of mdx-Na mice were reduced by 99% and were similar in control mice. These mdx mice displayed significantly elevated blood [Ca.sup.2+] and [Na.sup.+] levels, while the total calcium content of their TA muscle was reduced to the level of control mice. In addition, mdx-Na mice had elevated zinc and magnesium contents in their TA muscle. These results suggest that elevated serum [Na.sup.+] leads to [Ca.sup.2+] extrusion from muscle via the [Na.sup.+]/[Ca.sup.2+] exchanger causing a decrease in intracellular [Ca.sup.2+] levels and an increase in blood [Ca.sup.2+] levels. Extracellular [Ca.sup.2+] and, in addition, [Zn.sup.2+] and [Mg.sup.2+] might also contribute to the stabilization of the cell membrane. Other possibilities explaining the surprisingly efficacious beneficial effect of dietary sodium exist and are discussed. therapy; calcium and zinc; potassium; blood; serum creatine kinase activity

The gut hormone peptide YY(3-36) [PYY(3-36)] decreases food intake when administered by intravenous infusion to lean and obese humans and rats. Whether chronic administration of PYY(3-36) produces a sustained reduction in food intake and adiposity is the subject of intense debate. Batterham et al. (R. L. Batterham, M. A. Cowley, C. J. Small, H. Herzog, M. A. Cohen, C. L. Dakin, A. M. Wren, A. E. Brynes, M. J. Low, M. A. Ghatei, R. D. Cone, and S. R. Bloom. Nature 418: 650-654, 2002) first reported that PYY(3-36) reduces food intake and weight gain in rats when injected into the peritoneal cavity twice daily for 7 days. Numerous laboratories have failed to confirm that daily injections of PYY(3-36) decrease body weight. Continuous subcutaneous administration of PYY(3-36) by osmotic minipump has been reported to reduce daily food intake in rodents but only during the first 3-4 days of administration. Here we show the effects of different daily patterns of intravenous infusion of PYY(3-36) on food intake, body weight, and adiposity in rats tethered via infusion swivels to computer-controlled pumps. Measurement of food bowl weight recorded by computer every 20 s permitted daily assessment of the instantaneous effects of PYY(3-36) administration on food intake and meal patterns. One-hour intravenous infusions of PYY(3-36) at 30 pmol*[kg.sup.-1]*[min.sup.-1] every other hour for 10 days produced a sustained reduction in daily food intake of ~20% and decreased body weight and adiposity by 7 and 35%, respectively. Thus dosage pattern is critical for producing a sustained effect of PYY(3-36) on food intake and adiposity. gastrointestinal; body weight; body composition

Nonexercise activity thermogenesis (NEAT), the most variable component of energy expenditure, can account for differential capacities for human weight gain. Also highly variable, spontaneous physical activity (SPA) may similarly affect weight balance in animals. In the following study, we utilized the rat model of obesity, the diet-induced obese (DIO) rat, as well as the diet-resistant (DR) rat strain, to investigate how access to a high-fat diet alters SPA and the associated energy expenditure (i.e., NEAT). DIO and DR rats showed no differences in the amount of SPA before access to the high-fat diet. After 29 days on a high-fat diet, the DIO rats showed significant decreases in SPA, whereas the DR rats did not. Next, we wanted to determine whether the DIO and DR rats showed differential sensitivity to microinjections of orexin into the paraventricular nucleus of the hypothalamus (PVN). Unilateral guide cannulae were implanted, aimed at the PVN. Orexin A (0, 0.125, 0.25, and 1.0 nmol in 500 nl) was microinjected through the guide cannula into the PVN, then SPA and energy expenditure were measured for 2 h. Using the response to vehicle as a baseline, the DR rats showed significantly greater increase in NEAT compared with the DIO rats. These data indicate that diet-induced obesity is associated with decreases in SPA and a lack of increase in NEAT. A putative mechanism for changes in NEAT that accompany obesity is a decreased sensitivity to the NEAT-activating effects of neuropeptides such as orexin. nonexercise activity thermogenesis; paraventricular nucleus of the hypothalamus; high-fat diet; hypocretin

The objective of this study was to investigate meal-related endocrine changes that permit one to identify Sprague-Dawley rats at normal weight that are prone (OP) vs. resistant (OR) to obesity. In blood collected via chronic cardiac catheters, a 2-h high-fat meal (HFM, 50% fat, 40 kcal) at dark onset caused a significant increase in leptin, insulin, and triglycerides compared with premeal levels. Similar to patterns in already obese compared with lean rats on a high-fat diet, these meal-induced endocrine changes in normal-weight rats on lab chow were almost twofold larger in OP rats that, compared with OR rats, subsequently accumulated 100% more fat mass on a chronic high-fat diet. These exaggerated endocrine changes were similarly observed in blood collected using a simpler tail vein puncture procedure. In three separate experiments, the HFM-induced rise in leptin was found to be the strongest, positive correlate (r = +0.58, +0.62 and +0.64) of long-term body fat accrual. The lowest (2-5 ng/ml) vs. highest (6-9 ng/ml) scores for this post-HFM leptin measurement identified distinct OR and OP subgroups, respectively, when they were similar in body weight (340-350 g), premeal leptin (2.6-3.4 ng/ml), and meal size (40 kcal). Subsequent tests in these normal-weight OP rats revealed a distinct characteristic compared with OR rats, namely, exaggerated HFM-induced rise in expression of the orexigenic peptide galanin in the paraventricular nucleus. Thus, with this HFM-induced leptin measurement, OP rats can be identified while still at normal weight and then investigated for mechanisms that contribute to their excessive body fat accrual on a high-fat diet. insulin; triglycerides; galanin; neuropeptide Y

White-footed mice prey on gypsy moth pupae while foraging for other, more abundant food. Mice appear capable of locally extirpating moths since mice exert high predation pressure on sparse pupae and are numerically decoupled from moth populations. Nevertheless, during 23 years of monitoring, moths persisted at scales > 1 ha despite frequent extinctions at smaller spatial scales. We hypothesized that spatially heterogeneous intensity in mouse foraging and/or limited moth dispersal might allow moth persistence. Using a spatially explicit, individual-based, empirically parameterized model, we show that neither spatially heterogeneous predation by mice, nor limited moth dispersal alone allows moth persistence at typical mouse densities. However, synergy between both factors allows moth population persistence at naturally occurring mouse densities. For example, in models with 40 mice/ha, both limited moth dispersal with spatially homogeneous predation risk and spatially heterogeneous predation risk with unlimited moth dispersal had a 0% chance of moth persistence, but the combination of limited dispersal and heterogeneous predation risk resulted in a ~75% chance of moth persistence. Furthermore, both for limited moth dispersal with spatially homogeneous predation risk and for spatially heterogeneous predation risk with unlimited moth dispersal, moth persistence was only guaranteed at very low mouse densities, while the combination of limited moth dispersal with heterogeneous predation guaranteed moth persistence within a broad range of mouse densities. The findings illustrate a novel mechanism of 'spatial selection and satiation' that can enhance rare species persistence under intense incidental predation by generalist predators. Key words: gypsy moth; limited dispersal; Lymantria dispar; Peromyscus leucopus; population persistence; predation risk; spatial heterogeneity; white-footed mouse.

Rats acquired aversions to food pellets when a previously trained signal for that food was paired with a toxin, but only after minimal signal--food training. After extensive signal--food training, signal--toxin pairings had no effect on food consumption even after manipulations that enhanced the associability of the signal. By contrast, conditioned responding to the signal retained its sensitivity to devaluation of the food reinforcer by food--toxin pairings after extensive training. These results suggest that the nature of associatively activated event representations changes over the course of training.

Rats exposed to simultaneous compounds of 1 neutral flavor with dilute (2%) sucrose and a 2nd flavor with dilute (2%) maltodextrin subsequently consumed both flavors in preference to a 3rd flavor that was never paired with a palatable taste. Brief training exposure under ad lib food and water minimized the postingestive effects of nutrients, emphasizing the contribution of palatability to these preferences. Devaluation of sucrose or maltodextrin by pairing with illness (Experiment 1) or sensory-specific satiety (Experiment 2) selectively reduced the preference for the flavor previously paired with the devalued reinforcer. Such reinforcer-specific devaluation effects suggest that palatability-based learned flavor preferences are underpinned by a Pavlovian process whereby the cue flavor is associated with the taste of the concurrently consumed palatable reinforcer. Keywords: flavor preference learning, palatability, association, outcome

Five experiments investigated how rats' conditioned preferences or aversions for aqueous odors paired with sucrose or salt are affected by their unconditioned response to those tastes. Rats preferred an odor paired with 30% sucrose over an odor paired with 5% sucrose when both were presented in 5% sucrose, but they showed no preference or, if thirsty, showed the reverse preference, when the odors were presented in 30% sucrose. These changes in conditioned preference corresponded to changes in the rats' unconditioned preference for the accompanying sucrose solution. Rats' conditioned aversions for odors paired with salt showed a similar dependence on their reaction to the accompanying salt solution. The results were interpreted as showing that conditioned and unconditioned flavor preferences combine additively, as if mediated by the same sensory representation. Keywords: sucrose, salt, thirst, rat