Laboratory diet profoundly alters gene expression and confounds genomic analysis in mouse liver and lung

Nutritional studies in laboratory animals have long shown that various dietary components can contribute to altered gene expression and metabolism, but diet alone has not been considered in whole animal genomic studies. In this study, global gene expression changes in mice fed either a non-purified chow or a purified diet were investigated and background metal levels in the two diets were measured by ICP-MS. C57BL/6J mice were raised for 5 weeks on either the cereal-based, non-purified LRD-5001 diet or the purified, casein-based AIN-76A diet, as part of a larger study examining the effects of low dose arsenic (As) in the diet or drinking water. Affymetrix Mouse Whole Genome 430 2.0 microarrays were used to assess gene expression changes in the liver and lung. Microarray analysis revealed that animals fed the LRD-5001 diet displayed a significantly higher hepatic expression of Phase I and II metabolism genes as well as other metabolic genes. The LRD-5001 diet masked the As-induced gene expression changes that were clearly seen in the animals fed the AIN-76A diet when each dietary group was exposed to 100 ppb As in drinking water. Trace metal analysis revealed that the LRD-5001 diet contained a mixture of inorganic and organic As at a total concentration of 390 ppb, while the AIN-76A diet contained approximately 20 ppb. These findings indicate that the use of non-purified diets may profoundly alter observable patterns of change induced by arsenic and, likely, by other experimental treatments, particularly, altering gene and protein expression.     

Beneficial effect of goat milk on bioavailability of copper, zinc and selenium in rats

We studied the effects of dietary inclusion of freeze-dried goat and cow milk on the utilization of copper, zinc and selenium, and on the metabolic fate of copper and zinc, in rats using a standard (non-milk) control diet recommended by the American Institute of Nutrition and diets based on goat or cow milk. For animals given the goat milk diet, the apparent digestibility coefficient (ADC) of copper is similar to that obtained with the standard diet and higher than that in animals given the cow milk diet. The copper balance was higher among the rats given the goat milk and the standard diets than among those given cow milk. The ADC and retention of zinc and selenium were higher for the goat milk diet than for the other two diets. The copper content in the kidneys and in the femur was greater when the animals consumed a goat milk diet than a cow milk diet. Zn deposits in femur, testes, liver, kidney, heart and longissimus dorsi muscle were greatest with the goat-milk diet, followed by the standard diet and were lowest for the rats given cow-milk diet. This study shows that the goat-milk has an important and beneficial effect on the bioavailability of copper, zinc and selenium.     

Intracellular distribution and chemical forms of arsenic in rabbits exposed to arsenate

Inhibition of the methylation of arsenic in rabbits by ip injection of periodate-oxidized adenosine (PAD) prior to an iv injection of⁷⁴As-arsenate (AsV; 0.4 mg As/kg body wt) caused a marked increase in the retention of⁷⁴As in both the cellular organelles and the soluble fractions of liver and kidney. One day after exposure, almost 30% of the arsenic in the liver and about 40% of the arsenic in the kidney was recovered in the nuclear fraction. In the liver nuclei, the inhibition of the methylation increased the⁷⁴As content of the insoluble fraction and most of this arsenic was protein-bound. The major part of the soluble intranuclear⁷⁴As was in the form of AsIII, formed by reduction of the administered AsV. In the liver, PAD also caused a pronounced increase in the⁷⁴As content of the microsomal fraction. In the kidneys, where most of the arsenic was present as AsV, there was a marked accumulation of arsenic in the mitochondria.     

Dehydrogenase activity of liver parenchyma in mice exposed to arsenic in drinking water.

The effect of arsenic administered in drinking water in the form of sodium arsenite in concentrations of 5,50 and 250 mg As-3+/1 on dehydrogenase activity (DHA) of liver parenchyma homogenate of exposed mice was studied. After an exposure lasting 2, 4, 8, 16, 32, and 64 days, 8--10 exposed mice and 5--10 animals of the control series were studied at each interval. DHA was determined by means of tetrazolium chloride reduction to formazan whose quantity was assessed spectrophotometrically after extraction with butanol. At a concentration of 5 mg A3+/1 a moderate increase in DHA was observed which was replaced by a moderate decrease at the following intervals. At a concentration of 250 mg A3+/1 a deep decrease in DHA (p smaller than 0.05) was recorded which did not return to values compararable with those of the control series in spite of its fluctuating increase in the course of exposure. At a concentration of 50 mg A3+/1 in drinking water ensuring the supply of doses which induce an increase in the tolerance to arsenic a decrease in DHA was found only at the first exposure interval. DHA is thus substantially less altered in comparison with the metabolic consumption of oxygen (MCO) under the same conditions. This finding signifies that MCO is a more sensitive indicator of arsenic exposure than DHA in the used arrangement of the experiment.     

Distribution and Speciation of Arsenic by Transplacental and Early Life Exposure to Inorganic Arsenic in Offspring Rats

The amount of arsenic compounds was determined in the liver and brain of pups and in breast milk in the pup's stomach in relation to the route of exposure: transplacental, breast milk, or drinking water. Forty-eight pregnant rats were randomly divided into four groups, each group was given free access to drinking water that contained 0, 10, 50, and 100 mg/L NaAsO₂ from gestation day 6 (GD 6) until postnatal day 42 (PND 42). Once pups were weaned, they started to drink the same arsenic-containing water as the dams. Contents of inorganic arsenic (iAs), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), and trimethylarsenic acid (TMA) in livers and brains of the pups on PND 0, 15, 28, and 42 and breast milk taken from the pup's stomach on PND 0 and 15 were detected using the hydride generation atomic absorption spectroscopy method. Concentrations of iAs, MMA, and DMA in the breast milk, the brain, and the liver of the pups increased with the concentration of arsenic in drinking water on PND 0, 15, 28, and 42. Compared to the liver or brain, breast milk had the lowest arsenic concentrations. There was a significant decrease in the levels of arsenic species on PND 15 compared to PND 0, 28, or 42. It was confirmed that arsenic species can pass through the placental barrier from dams to offspring and across the blood–brain barrier in the pups, and breast milk from dams exposed to arsenic in drinking water contains less arsenic than the liver and brain of pups.     

High dietary fat exacerbates arsenic-induced liver fibrosis in mice

Many factors could potentially affect the process of arsenic-induced liver fibrosis. The present study was undertaken to examine the effect of high fat diet on arsenic-induced liver fibrosis and preneoplastic changes. Mice were given sodium arsenite (As3+, 200 ppm) or sodium arsenate (As5+, 200 ppm) in the drinking water for 10 months, and provided a normal diet or a diet containing 20% added fat. Serum aspartate aminotransferase (AST), indicative of liver injury, was elevated in both arsenite and arsenate groups, and a high fat diet further increased these levels. Histopathology (H&E and Masson stain) showed that liver inflammation, steatosis (fatty liver), hepatocyte degeneration, and fibrosis occurred with arsenic alone, but their severity was markedly increased with the high fat diet. Total liver RNA was isolated for real-time RT-PCR analysis. Arsenic exposure increased the expression of inflammation genes, such as TNF-alpha, IL-6, iNOS, chemokines, and macrophage inflammatory protein-2. The expression of the stress-related gene heme oxygenase-1 was increased, while metallothionein-1 and GSH S-transferase-pi were decreased when arsenic was combined with the high fat diet. Expression of genes related to liver fibrosis, such as procollagen-1 and -3, SM-actin and TGF-beta, were synergistically increased in the arsenic plus high fat diet group. The expression of genes encoding matrix metalloproteinases (MMP2, MMP9) and tissue inhibitors of metalloproteinases (TIMP1, TIMP2) was also enhanced, suggestive of early oncogenic events. In general, arsenite produced more pronounced effects than arsenate. In summary, chronic inorganic arsenic exposure in mice produces liver injury, and a high fat diet markedly increases arsenic-induced hepatofibrogenesis.     

Effect of arsenic trioxide on metallothionein and its conversion to different arsenic metabolites in hen liver

The metabolism of arsenic, its affinity to metallothionein (MT), its influence on selenium levels, and its biotransformation to different metabolites in the liver tissue of laying hens exposed to arsenic trioxide (As2O3) was investigated. The experiment was performed with two groups of hens fed for 19 d with either a standard diet or with the same diet enriched in arsenic (30 microg/g). The major findings were as follows: 1. After 19 d exposure, about 65% of the total liver As was found in the water-soluble phase (100,000g centrifuged supernatant). In liver supernatant, As binding was found mostly in the range of very low-molecular-weight proteins (Mr < 10,000). Although after exposure the amount of MT-like proteins increased, the As bound to it was only in trace amounts. The protein was identified by convential procedures as Zn,Cu-thionein with traces of selenium and arsenic. 2. Arsenic exposure resulted in almost unchanged Se levels regarding its tissue concentrations and distribution between supernatant and pellet, where about 10% of total Se was found in the supernatant. On the contrary, As exposure did affect Cd levels. Tissue Cd concentration was slightly diminished, but the percentage of tissue Cd found in the water-soluble phase was increased from 20% to 40%. 3. In methanol extracts of tissue and supernatant of the As-exposed group, only two arsenic compounds were detected, As(III) and dimethylarsinic acid (DMA), the latter prevailing.     

Effect of long-term administration of arsenic (III) and bromine with and without selenium and iodine supplementation on the element level in the thyroid of rat

The aim of this study was to evaluate the influence of arsenic and bromine exposure with or without iodine and selenium supplementation on the element level in the thyroid of rats. Four major groups of Wistar female rats were fed with respective diets: group A - standard diet, group B - iodine rich diet (10 mg I/kg food), group C - selenium rich diet (1 mg Se/kg) and group D - iodine and selenium rich diet (as in group B and C). Each group was divided into four subgroups per 7 animals each receiving either NaAsO(2) ip (6.5 mg.kg(-1) twice a week for two weeks and 3.25 mg.kg(-1) for six weeks) or KBr in drinking water (58.8 mg.l(-1)) for 8 weeks or combined administration of both substances. Remaining subgroup served as controls. After 8 weeks thyroid glands were analyzed by ICP-MS for As, Br, Se, and I content. The exposition of rat to arsenic or bromine causes the accumulation of these elements in the thyroid gland ( approximately 18 ppm of As, approximately 90 ppm of Br) and significantly affects iodine and selenium concentration in the thyroid. In iodine and/or selenium supplemented rats the bromine intake into the thyroid was lowered to approximately 50% of the level in unsupplemented animals. Also selenium thyroid level elevated due to KBr administration was lowered by iodine supplementation in the diet. The accumulation of arsenic in the thyroid was not influenced by selenium or iodine supplementation; however, As(III) administration increased iodine thyroid level and suppressed selenium thyroid level in selenium or iodine supplemented group of animals.     

Oxidative stress in mice is dependent on the free glucose content of the diet

In animals, chronic intake of diets with high proportions of rapidly absorbable glucose promotes the development of insulin resistance. High levels of glucose can produce permanent chemical alterations in proteins and lipid peroxidation. delta-Aminolevulinate dehydratase (delta-ALA-D) is a sulfhydryl-containing enzyme essential for all aerobic organisms and is highly sensitive to the presence of pro-oxidants elements. The heme synthetic pathway is impaired in porphyria and a frequent coexistence of diabetes mellitus and porphyria disease has been reported in humans and experimental animal models, which can be casually linked to the delta-ALA-D inhibition found in diabetics. The present study was designed to evaluate the effect of two different diets, a high glucose (HG) diet and a high starch (HS) diet, on lipid peroxidation levels in different tissues (brain, liver, and kidney) and on delta-ALA-D activity (from liver and kidney) in mice. Plasma glucose and triglyceride levels were significantly higher in mice fed HG than in mice fed HS (P < 0.02 and P < 0.03, respectively). Thiobarbituric acid reactive species (TBA-RS) content was significantly increased in kidney and liver from HG diet-fed mice when compared with animals fed HS diets (P < 0.001). Hepatic delta-ALA-D activity of HG diet-fed animals was significantly lower than that of HS diet-fed animals (P < 0.01). The results of this study support the hypothesis that consumption of a diet with high free glucose can promote the development of oxidative stress that we tentatively attribute to hyperglycemia.     

Noncancer risk assessment of arsenic safe and unsafe water uses: Bayesian estimation on cohort study

Cancer and noncancer risk of arsenic exposure depends on arsenic intake through drinking water and diets. The present study evaluated the probability of noncancer effects of arsenic exposure from drinking water and diets in a cohort of 82 participants in arsenic-endemic rural areas, considering arsenic-safe and arsenic-unsafe water uses for three consecutive years. The risk assessment included the collection of last 24 hours' diet replica and urine of the participants followed by total arsenic analysis of the same. Toxic dose emerging from exposure duration is a nonlinear variable. So, Bayesian estimation of the data for noncancer risk assessment of the variable arsenic consumption was performed. In spite of using arsenic-safe water, we observed arsenic consumption and release. Participants with skin lesions had more arsenic in urine than participants without skin lesions. Future risk for participants without skin lesions was twice due to less arsenic release in urine. For the first time, Bayesian simulation was used to assess noncancer risk on a cohort for a consecutive three-year study. A significant finding was the higher assessed noncancer risk of the participants without skin lesions than the participants with skin lesions.     

A Review on the Role of Chromium Supplementation in Ruminant Nutrition—Effects on Productive Performance,Blood Metabolites,Antioxidant Status,and Immunocompetence

Interactions of arsenic with essential trace elements may result in disturbances on body homeostasis. In the present study, we aimed to investigate the effects of different arsenic compounds on micromineral content and antioxidant enzyme activities in rat liver. Male Wistar rats were randomly divided into five groups and exposed to sodium arsenite and sodium arsenate at 0.01 and 10 mg/L for 8 weeks in drinking water. The concentration of arsenic increased in the liver of all arsenic-exposed animals. The proportion of zinc and copper increased in animals exposed to 0.01 mg/L sodium arsenite. In addition, these animals presented a reduction in magnesium and sodium content. Superoxide dismutase activity decreased mainly in arsenite-exposed animals, whereas catalase activity decreased in animals exposed to 10 mg/L sodium arsenate. Further, exposure to sodium arsenate at 10 mg/L altered copper and magnesium content in the liver, and reduced total protein levels. Overall, both arsenic compounds altered the liver histology, with reduction in the proportion of cytoplasm and hepatocyte, and increased the percentage of sinusoidal capillaries and macrophages. In conclusion, our findings showed that oral exposure to arsenic compounds disturbs the trace elements balance in the liver, especially at low concentration, altering enzymatic and stereological parameters. We concluded that despite the increase in trace elements content, the antioxidant enzyme activities were downregulated and did not prevent morphological alterations in the liver of animals exposed to both arsenic compounds.     

Incorporation of C14-thymidine and C14-uridine nucleosides into nucleic acid molecules in the liver of mice exposed to trivalent arsenic in drinking water

The effect of trivalent arsenic on the C14-thymidine and C14-uridine incorporation into the liver nucleic acid molecules was studied in groups of mice exposed to 3.3 mmol . l-1 sodium arsenite in drinking water for 2, 4, 8, 16 and 32 days. In each exposure group, 18 h before the animals were killed by decapitation, six to ten mice were injected with 0.1 ml of C14-thymidine or C14-uridine of an activity of 0.37 MBq. The radioactivity of liver DNA and liver RNA fractions isolated and separated by extraction from the mouse liver homogenate was measured on a Betazcint BZ 5,000 scintillation counter and expressed as cpm . mg-1 of isolated nucleic acids. Initially, the rates at which the labeled thymidine and uridine nucleosides incorporated into the nucleic acid molecules showed a marked drop below the control value, then started to rise again, continuing to gradually increase as the time of arsenic exposure was increasing; they attained the control value on day 8, and at the last interval of observation (i.e. at 32 days of exposure to arsenic) they reached, respectively, 143% and 127% of the control value. The initial drop in the rate of nucleoside incorporation is believed to reflect the general metabolic inhibition caused by arsenic; the subsequent enhancement of the process of incorporation is ascribed to the induction of enzymes that are involved in the development of a tolerance for arsenic.     

Ornithine decarboxylase and polyamines in liver and kidneys of rats on cyclical regimen of protein-free and protein-containing diets. Relationship to deoxyribonucleic acid synthesis in liver

1. The activity of ornithine decarboxylase in the liver and kidneys of rats maintained on a cyclical regimen of protein-free and protein-containing diets was investigated. There was a daily activation of the enzyme in response to the feeding of protein after 3 days feeding of protein-free diet. 2. The activation of ornithine decarboxylase in the liver and kidneys of rats re-fed on protein was demonstrable throughout 16 cycles of alternating 3-day periods of protein-free and protein-containing diets. The magnitude of the activation in the kidneys diminished from 20-fold stimulation in the first cycle to 5-fold stimulation (compared with animals fed with protein-free diet) in the later cycles of protein re-feeding. The activation of the enzyme in liver was decreased from 20-fold stimulation in the first cycle to approx. 10-fold stimulation in later cycles. 3. The concentration of spermidine was increased by approx. 50% in the liver of animals during cycling from protein-free to protein-containing diets. Spermine was unchanged, and putrescine was maintained at a low concentration approx. one-fifth to one-tenth that of spermidine after protein re-feeding. 4. The incorporation of [(3)H]thymidine into liver DNA was increased 10-fold in animals re-fed with protein compared with animals receiving protein-free diets. 5. The activation of ornithine decarboxylase by re-feeding of protein was inhibited 90% by the injection of propane-1,3-diamine during re-feeding. The stimulation of DNA synthesis was inhibited 60% by multiple injections of propane-1,3-diamine during the re-feeding of protein.     

Importance of Arsenic Speciation in Populations Exposed to Arsenic in Drinking Water

Total arsenic in urine is often the principal means for assessing chronic exposure to arsenic-contaminated drinking water. This approach ignores many components of the human diet, especially fish and seafood that contain arsenic at significant concentrations. The toxicity differences between the inorganic forms and the dietary forms suggest both should be evaluated when attempting to assess risk from arsenic exposure. Urine biomonitoring for 53 participants was used to confirm reduction in arsenic exposure resulting from well water remediation removing inorganic arsenic from drinking water. Initially, only total arsenic urine assays were performed, but spikes in total arsenic urine concentrations were determined to be diet related and demonstrated the need for analytical methods that differentiate the arsenic species. A secondary analysis was added that quantified inorganic-related arsenic in urine and the dietary forms related to fish and seafood by subtraction from total arsenic. Significant differences were found between the inorganic arsenic component and the total arsenic measured in their urine. On average, approximately 76% of total arsenic in urine was attributed to fish and other organo-arsenic dietary sources, implying a potential significant overestimate of exposure, and demonstrating the need for differentiation of the inorganic-related arsenic from dietary arsenic.     

Effects of dietary mulberry, Korean red ginseng, and banaba on glucose homeostasis in relation to PPAR-alpha, PPAR-gamma, and LPL mRNA expressions

Despite lack of scientific evidences to support its therapeutic efficacy, the use of herbal supplements has significantly increased. The purpose of this study was to evaluate the effects of traditional anti-diabetic herbs on the progress of diabetes in db/db mice, a typical non-insulin-dependent model. Five different experimental diets were as follows: control diet, 0.5% mulberry leaf water extract diet, 0.5% Korean red ginseng diet, 0.5% banaba leaf water extract diet, and 0.5% combination diet (mulberry leaf water extract/Korean red ginseng/banaba leaf water extract, 1:1:1). Blood levels of glucose, insulin, HbA1c, and triglyceride were measured every 2 weeks. At 12 weeks of age, animals were sacrificed, and tissue mRNA levels of PPAR-alpha, PPAR-gamma, and LPL were determined. Results indicated that mulberry leaf water extract, Korean red ginseng, banaba leaf water extract, and the combination of above herbs effectively reduced blood glucose, insulin, TG, and percent HbA1c in study animals (p<0.05). We also observed that the increased expressions of liver PPAR-alpha mRNA and adipose tissue PPAR-gamma mRNA in animals fed diets supplemented with test herbs. The expression of liver LPL mRNA was also increased with experimental diets containing herbs. The efficacy was highest in animals fed the combination diet for all of the markers used. These results suggest that mulberry leaf water extract, Korean red ginseng, banaba leaf water extract, and the combination of these herbs fed at the level of 0.5% of the diet significantly increase insulin sensitivity, and improve hyperglycemia possibly through regulating PPAR-mediated lipid metabolism.     

The effect of dietary protein on the mineral status of vervet monkeys with special reference to the impact of milk solids on calcium excretion

This study assessed the impact of Westernised and traditional African diets on mineral metabolism in general and calcium status in particular in vervet monkeys. Twelve adult male vervet monkeys (Cercopithecus aethiops), with an average weight of 5+/-0.58 kg each, were divided into two groups of six individuals each and fed traditional diets containing largely maize + legumes (17.4%) or Westernised diets containing milk solids (17.2%) as the source of high crude protein for 8 weeks. Blood was taken at 2-week intervals, the animals were weighed, while urine and stool samples were collected over 24 hours. The monkeys on the milk solids diet had diarrhoea for 6 weeks post-dietary intervention, and produced significantly greater quantities (P<0.02) of stool. These animals also produced significantly more urine (P<0.02). There was no difference in the degree of calciuresis of the two groups, but the monkeys on maize + legume proteins absorbed significantly more calcium during weeks two and six (P<0.04). Furthermore, both groups of monkeys showed a significant decline in plasma calcium levels over the experimental period (P<0.001). The diets had no effect on phosphate levels in the plasma or urine. However, both groups of animals absorbed less phosphate (P<0.09). There was an increasing loss of urinary magnesium (P = 0.03) in both groups, with the milk solids group showing lower plasma levels of this element (P = 0.09). However, the milk solids group lost less magnesium through the stool (P<0.03). In addition, the animals on milk solids showed significant natriuresis (P<0.05), while plasma sodium levels in both groups declined over time (P<0.03). Both diets induced a state of urinary potassium loss (P = 0.0003) and decrease in plasma potassium (P<0.0002). Urinary pH and plasma urea were unaffected by the diets, but the monkeys on maize + legumes excreted significantly less (P<0.001) urinary urea. This study indicates that the milk solids diet compromised mineral homeostasis by interfering with gut and renal functioning, while the traditional African diet did not induce these effects.     

Dietary methionine effects on plasma homocysteine and HDL metabolism in mice

The effects of dietary manipulation of folate and methionine on plasma homocysteine (Hcy) and high-density lipoprotein cholesterol (HDL-C) levels in wild-type and apolipoprotein-E-deficient mice were determined. A low-folate diet with or without folate and/or methionine supplementation in drinking water was administered for 7 weeks. Fasted Hcy rose to 23 microM on a low-folate/high-methionine diet, but high folate ameliorated the effect of high methionine on fasted plasma Hcy to approximately 10 microM. Determination of nonfasted plasma Hcy levels at 6-h intervals revealed a large diurnal variation in Hcy consistent with a nocturnal lifestyle. The daily average of nonfasted Hcy levels was higher than fasted values for high-methionine diets but lower than fasted values for low-methionine diets. An acute methionine load by gavage of fasted mice increased plasma Hcy 2.5 h later, but mice that had been on high-methionine diets had a lower fold induction. Mice fed high-methionine diets weighed less than mice fed low-methionine diets. Based on these results, two solid-food diets were developed: one containing 2% added methionine and the other containing 2% added glycine. The methionine diet led to fasted plasma Hcy levels of >60 microM, higher than those with methionine supplementation in drinking water. Mice on methionine diets had >20% decreased body weights and decreased HDL-C levels. An HDL turnover study demonstrated that the HDL-C production rate was significantly reduced in mice fed the methionine diet.     

Overeating of palatable food is associated with blunted leptin and ghrelin responses

Palatable food is rich in fat and/or sucrose. In this study we examined the long-term effects of such diets on food intake, body weight, adiposity and circulating levels of the satiety peptide leptin and the hunger peptide ghrelin. The experiments involved rats and mice and lasted 5 weeks. In rats, we examined the effect of diets rich in fat and/or sucrose and in mice the effect of a high fat diet with or without sucrose in the drinking water. Animals fed with the palatable diets had a larger intake of calories, gained more weight and became more adipose than animals fed standard rat chow. Fasted animals are known to have low serum leptin and high serum ghrelin and to display elevated serum leptin and lowered serum ghrelin postprandially. With time, a sucrose-rich diet was found to raise the fasting level of leptin and to lower the fasting level of ghrelin in rats. A fat-rich diet suppressed serum ghrelin without affecting serum leptin; high sucrose and high fat in combination greatly reduced serum ghrelin and raised serum leptin in the fasted state. The mRNA expression of leptin in the rat stomach was up-regulated by sucrose-rich (but not by fat-rich) diets, whereas the expression of ghrelin seemed not to be affected by the palatable diets. Mice responded to sucrose in the drinking water with elevated serum leptin (fasted state) and to all palatable diets with low serum ghrelin. The expression of both leptin and ghrelin mRNA in the stomach was suppressed in fasted mice that had received a high fat diet for 5 weeks. We conclude that the expression of leptin mRNA in stomach and the concentration of leptin in serum were elevated in response to sucrose-rich rather than fat-rich diets, linking leptin with sucrose metabolism. In contrast, the expression of ghrelin and the serum ghrelin concentration were suppressed by all palatable diets, sucrose and fat alike. In view of the increased body weight and adiposity neither elevated leptin nor suppressed ghrelin were able to control/restrain the overeating that is associated with palatable diets.     

Effects of individual and combined exposure to sodium arsenite and sodium fluoride on tissue oxidative stress, arsenic and fluoride levels in male mice

Arsenic and fluoride are potent toxicants, widely distributed through drinking water and food and often result in adverse health effects. The present study examined the effects of sodium meta-arsenite (100 mg/l in drinking water) and sodium fluoride (5 mg/kg, oral, once daily), administered either alone or in combination for 8 weeks, on various biochemical variables indicative of tissue oxidative stress and cell injury in Swiss albino male mice. A separate group was first exposed to arsenic for 4 weeks followed by 4 weeks of fluoride exposure. Exposure to arsenic or fluoride led to a significant depletion of blood delta-aminolevulinic acid dehydratase (ALAD) activity and glutathione (GSH) level. These changes were accompanied by increased level of blood and tissues reactive oxygen species (ROS) level. An increase in the level of liver and kidney thiobarbituric acid reactive substance (TBARS) along with a concomitant decrease in the activities of superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx) and reduced GSH content were observed in both arsenic and fluoride administered mice. The changes were significantly more pronounced in arsenic exposed animals than in fluoride. It was interesting to observe that during combined exposure the toxic effects were less pronounced compared to the effects of arsenic or fluoride alone. In some cases antagonistic effects were noted following co-exposure to arsenic and fluoride. Arsenic and fluoride concentration increased significantly on exposure. Interestingly, their concentration decreased significantly on concomitant exposure for 8 weeks. However, the group which was administered arsenic for 4 weeks followed by 4 weeks of fluoride administration showed no such protection suggesting that the antagonistic effect of fluoride on arsenic or vice versa is possible only during interaction at the gastro intestinal sites. These results are new and interesting and require further exploration.     

Utilization by sheep of dried pig faeces containing a high concentration of copper

Seven groups of three sheep were given pelleted diets as follows: A, 97% hay + 3% molasses; B, C, D, 82% hay + 3% molasses + 15% dried pig faeces; E, F, G, 67% hay + 3% molasses + 30% dried pig faeces. In an attempt to counteract potential toxic effects of copper in the pig faeces (613 mg/kg dry matter), molybdenum was added to diets C and F at the rate of 90 mg/kg diet, and to diets D and G at the rate of 175 mg/kg diet. Sulphate was included in all diets at the rate of 1.08% of diet. Measurements were made of digestibility, copper retention, and plasma glutamic oxaloacetic transaminase (GOT) values during a 70-day feeding period, after which the animals were slaughtered.Dry matter digestibility coefficients were : diet A, 49.2; diets B, C, D, 44.4; diets E, F, G, 42.5, from which the coefficient for pig faeces alone is calculated to be < 30. Mean copper retention over the 70 days ranged from 210 mg for diet A to 1225 mg for diet G (excluding diet E which showed an aberrant retention of 53 ± 185 mg). Plasma GOT values fluctuated between diets and between sheep with a maximum value of 182 units/ml. Mean liver copper concentrations ranged from 718 mg/kg dry matter for diet A to 1740 mg/kg for diet E. Necrotic lesions occurred in some livers. Kidneys were apparently normal. There were no clear differences in copper status of animals receiving 15% versus 30% pig faeces except for a tendency for liver damage to be greater in the latter. There were no apparent effects of additions of molybdenum.The pig faeces were poorly utilized and, because of the high copper content, potentially hazardous to the health of the sheep.