The Protective Effect of Nutraceuticals on Hepatic Ischemia-Reperfusion Injury in Wistar Rats.

Nutraceuticals are bioactive compounds present in foods, utilized to ameliorate health, prevent diseases, and support the proper functioning of the human body. They have gained attention due to their ability to hit multiple targets and act as antioxidants, anti-inflammatory agents, and modulators of immune response and cell death. Therefore, nutraceuticals are being studied to prevent and treat liver ischemia-reperfusion injury (IRI). This study evaluated the effect of a nutraceutical solution formed by resveratrol, quercetin, omega-3 fatty acid, selenium, ginger, avocado, leucine, and niacin on liver IRI. IRI was performed with 60 min of ischemia and 4 h of reperfusion in male Wistar rats. Afterward, the animals were euthanized to study hepatocellular injury, cytokines, oxidative stress, gene expression of apoptosis-related genes, TNF-α and caspase-3 proteins, and histology. Our results show that the nutraceutical solution was able to decrease apoptosis and histologic injury. The suggested mechanisms of action are a reduction in gene expression and the caspase-3 protein and a reduction in the TNF-α protein in liver tissue. The nutraceutical solution was unable to decrease transaminases and cytokines. These findings suggest that the nutraceuticals used favored the protection of hepatocytes, and their combination represents a promising therapeutic proposal against liver IRI.

Echium oil with oxidative stability increased by emulsion preparation in the presence of the phenolic compound sinapic acid followed by dehydration by spray and freeze drying processes.

Echium oil is rich in omega-3, however, is unstable. The objective of this work was the co-encapsulation of echium oil and sinapic acid (SA) by emulsification using Arabic gum as emulsifier/carrier, followed by spray or freeze-drying. Eight treatments (S0, S200, S600 and S1000: particles spray dried with different concentrations of SA; L0, L200, L600 and L1000: particles freeze dried with different concentrations of SA) were analyzed in relation to microscopy, water activity (Aw), hygroscopicity, moisture, solubility, particle size, X-ray diffraction, thermogravimetry and accelerated oxidation. Particles of rounded shape and undefined form were obtained by spray and freeze-drying, besides ideal physicochemical properties for application (values from 0.091 to 0.365, 3.22 to 4.89%, 57 to 68% and 2.32 to 12.42 µm for Aw, moisture, solubility and particle size, respectively). All treatments protected the oil against oxidation, obtaining induction time of 5.31 h for oil and from 7.88 to 12.94 h for treatments. The better protection to oil was obtained with it emulsified and freeze-dried (L600); the encapsulation increased oxidative stability of the oil, besides facilitating its application over the fact the material is in powder form.

Protective effect of genetic deletion of pannexin1 in experimental mouse models of acute and chronic liver disease.

Pannexins are transmembrane proteins that form communication channels connecting the cytosol of an individual cell with its extracellular environment. A number of studies have documented the presence of pannexin1 in liver as well as its involvement in inflammatory responses. In this study, it was investigated whether pannexin1 plays a role in acute liver failure and non-alcoholic steatohepatitis, being prototypical acute and chronic liver pathologies, respectively, both featured by liver damage, oxidative stress and inflammation. To this end, wild-type and pannexin1-/- mice were overdosed with acetaminophen for 1, 6, 24 or 48h or were fed a choline-deficient high-fat diet for 8weeks. Evaluation of the effects of genetic pannexin1 deletion was based on a number of clinically relevant read-outs, including markers of liver damage, histopathological analysis, lipid accumulation, protein adduct formation, oxidative stress and inflammation. In parallel, in order to elucidate molecular pathways affected by pannexin1 deletion as well as to mechanistically anchor the clinical observations, whole transcriptome analysis of liver tissue was performed. The results of this study show that pannexin1-/- diseased mice present less liver damage and oxidative stress, while inflammation was only decreased in pannexin1-/- mice in which non-alcoholic steatohepatitis was induced. A multitude of genes related to inflammation, oxidative stress and xenobiotic metabolism were differentially modulated in both liver disease models in wild-type and in pannexin1-/- mice. Overall, the results of this study suggest that pannexin1 may play a role in the pathogenesis of liver disease.

Bioactive compounds as an alternative for drug co-therapy: Overcoming challenges in cardiovascular disease prevention.

Different pharmacological interventions have been applied with success to reduce the progression of atherosclerosis. However, many patients are not good responders or must interrupt treatment due to adverse effects. Bioactive compounds such as omega-3 fatty acids (n-3 FA), plant sterol esters (PSE) and phenolic compounds (PHC) are natural molecules with great potential to reduce the atherosclerosis burden by reducing inflammation, LDL cholesterol (LDL-C) and oxidative stress, respectively. Although their physiological effects on biomarkers are much lower than those expected by drugs used for the same purpose, bioactive compounds can easily be incorporated into the daily diet and present no adverse effects. However, little is known about the combination of n-3 FA, PSE, PHC, and drugs in atherosclerosis progression. This review article summarizes potential effects of co-therapies involving n-3 FA, PSE, and PHC combined with major hypolipidemic drugs on atherosclerosis biomarkers and clinical outcomes. Evidence of additive and/or complementary effects regarding drugs action reveals possible roles for bioactive compounds in disease management. Pharmaceutical companies, physicians, and food scientists should be prepared to better understand this type of interaction and its consequences in terms of efficacy and life quality.

Connexin32 deficiency is associated with liver injury, inflammation and oxidative stress in experimental non-alcoholic steatohepatitis.

Non-alcoholic steatohepatitis is a highly prevalent liver pathology featured by hepatocellular fat deposition and inflammation. Connexin32, which is the major building block of hepatocellular gap junctions, has a protective role in hepatocarcinogenesis and is downregulated in chronic liver diseases. However, the role of connexin32 in non-alcoholic steatohepatitis remains unclear. Connexin32-/- mice and their wild-type littermates were fed a choline-deficient high-fat diet. The manifestation of non-alcoholic steatohepatitis was evaluated based on a battery of clinically relevant read-outs, including histopathological examination, diverse indicators of inflammation and liver damage, in-depth lipid analysis, assessment of oxidative stress, insulin and glucose tolerance, liver regeneration and lipid-related biomarkers. Overall, more pronounced liver damage, inflammation and oxidative stress were observed in connexin32-/- mice compared to wild-type animals. No differences were found in insulin and glucose tolerance measurements and liver regeneration. However, two lipid-related genes, srebf1 and fabp3, were upregulated in Cx32-/- mice in comparison with wild-type animals. These findings suggest that connexin32-based signalling is not directly involved in steatosis as such, but rather in the sequelae of this process, which underlie progression of non-alcoholic steatohepatitis.

Paternal programming of breast cancer risk in daughters in a rat model: opposing effects of animal- and plant-based high-fat diets.

BACKGROUND: Although males contribute half of the embryo's genome, only recently has interest begun to be directed toward the potential impact of paternal experiences on the health of offspring. While there is evidence that paternal malnutrition may increase offspring susceptibility to metabolic diseases, the influence of paternal factors on a daughter's breast cancer risk has been examined in few studies. METHODS: Male Sprague-Dawley rats were fed, before and during puberty, either a lard-based (high in saturated fats) or a corn oil-based (high in n-6 polyunsaturated fats) high-fat diet (60 % of fat-derived energy). Control animals were fed an AIN-93G control diet (16 % of fat-derived energy). Their 50-day-old female offspring fed only a commercial diet were subjected to the classical model of mammary carcinogenesis based on 7,12-dimethylbenz[a]anthracene initiation, and mammary tumor development was evaluated. Sperm cells and mammary gland tissue were subjected to cellular and molecular analysis. RESULTS: Compared with female offspring of control diet-fed male rats, offspring of lard-fed male rats did not differ in tumor latency, growth, or multiplicity. However, female offspring of lard-fed male rats had increased elongation of the mammary epithelial tree, number of terminal end buds, and tumor incidence compared with both female offspring of control diet-fed and corn oil-fed male rats. Compared with female offspring of control diet-fed male rats, female offspring of corn oil-fed male rats showed decreased tumor growth but no difference regarding tumor incidence, latency, or multiplicity. Additionally, female offspring of corn oil-fed male rats had longer tumor latency as well as decreased tumor growth and multiplicity compared with female offspring of lard-fed male rats. Paternal consumption of animal- or plant-based high-fat diets elicited opposing effects, with lard rich in saturated fatty acids increasing breast cancer risk in offspring and corn oil rich in n-6 polyunsaturated fatty acids decreasing it. These effects could be linked to alterations in microRNA expression in fathers' sperm and their daughters' mammary glands, and to modifications in breast cancer-related protein expression in this tissue. CONCLUSIONS: Our findings highlight the importance of paternal nutrition in affecting future generations' risk of developing breast cancer.

Yerba Mate (Ilex paraguariensis) modulates NF-kappaB pathway and AKT expression in the liver of rats fed on a high-fat diet.

To investigate the effect of Yerba Mate (YM) aqueous extract intake on the NF-kB pathway and AKT expression in the liver, muscle, and adipose tissue of rats submitted to a high-fat diet (HFD). Male Wistar rats were fed a control (CON) (n = 24) or a HFD (n = 24) for 12 weeks. Afterwards, rats received YM daily (1 g/kg body weight) for 4 weeks. Intake of YM aqueous extract reduced body weight gain (p < 0.05) and total blood cholesterol (p < 0.05) in the HFD group in comparison to the non-treated HFD group. HFD group demonstrated an increased glycemic response at 5 and 10 min after insulin injection. YM decreased the ratio between phosphorylated and total kinase inhibitor of κB (IKK), increased the ratio of phosphorylated to total form of protein kinase B (AKT) and reduced NF-κB phosphorylation in the liver of the HFD group. Our data suggest a beneficial role of YM in improving metabolic dysfunctions induced by HFD.

Effect of Echium oil compared with marine oils on lipid profile and inhibition of hepatic steatosis in LDLr knockout mice.

BACKGROUND: In an effort to identify new alternatives for long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) supplementation, the effect of three sources of omega 3 fatty acids (algae, fish and Echium oils) on lipid profile and inflammation biomarkers was evaluated in LDL receptor knockout mice. METHODS: The animals received a high fat diet and were supplemented by gavage with an emulsion containing water (CON), docosahexaenoic acid (DHA, 42.89%) from algae oil (ALG), eicosapentaenoic acid (EPA, 19.97%) plus DHA (11.51%) from fish oil (FIS), and alpha-linolenic acid (ALA, 26.75%) plus stearidonic acid (SDA, 11.13%) from Echium oil (ECH) for 4 weeks. RESULTS: Animals supplemented with Echium oil presented lower cholesterol total and triacylglycerol concentrations than control group (CON) and lower VLDL than all of the other groups, constituting the best lipoprotein profile observed in our study. Moreover, the Echium oil attenuated the hepatic steatosis caused by the high fat diet. However, in contrast to the marine oils, Echium oil did not affect the levels of transcription factors involved in lipid metabolism, such as Peroxisome Proliferator Activated Receptor α (PPAR α) and Liver X Receptor α (LXR α), suggesting that it exerts its beneficial effects by a mechanism other than those observed to EPA and DHA. Echium oil also reduced N-6/N-3 FA ratio in hepatic tissue, which can have been responsible for the attenuation of steatosis hepatic observed in ECH group. None of the supplemented oils reduced the inflammation biomarkers. CONCLUSION: Our results suggest that Echium oil represents an alternative as natural ingredient to be applied in functional foods to reduce cardiovascular disease risk factors.

The effect of mate tea (Ilex paraguariensis) on metabolic and inflammatory parameters in high-fat diet-fed Wistar rats.

This study investigated the effects of mate tea (Ilex paraguariensis) aqueous extract consumption on metabolic indicators and inflammatory response of peritoneal macrophages in rats fed a high-fat diet (HFD). Male Wistar rats were fed a control diet or a HFD for 12 weeks. At the end of this period, rats received, or not, daily doses of yerba maté for 4 weeks. The consumption of yerba maté promoted weight loss, attenuated the HFD-detrimental effects on adiposity and insulin sensitivity and decreased blood levels of the inflammatory biomarkers (p < 0.05). Concerning peritoneal macrophages, mate tea consumption decreased the production of interleukin (IL)-6, but did not influence the production of IL-1ß, tumour necrosis factor-α and nitric oxide; cytokine mRNA expression; or the activation of the nuclear factor-κB signalling pathway. In summary, the consumption of mate tea had no consistent effect in the inflammatory response of peritoneal macrophages, but reduced cardiometabolic risk markers.

Could a lipid oxidative biomarker be applied to improve risk stratification in the prevention of cardiovascular disease?

There is significant evidence demonstrating the influence of oxidative stress on atherosclerosis and cardiovascular diseases (CVD). However, oxidative biomarkers have not been applied to follow patients under primary or secondary prevention. Many factors can explain this paradox: the higher complexity of the methods applied to quantify oxidative markers, the high variability observed among the studies, the lack of reference values, and the weak correlation with clinical endpoints. This review presents the role of the major reactive oxygen species (ROS) involved in cardiovascular pathophysiology and how they can be neutralized by endogenous and exogenous antioxidants based on classical and recent studies, highlighting the importance of the secondary products of fatty acid oxidation as potential biomarkers. Furthermore, we discuss the great variability of oxidative stress biomarkers, using as an example data obtained from 55 studies. Among the molecules directly formed from lipid oxidation, such as malondialdehyde (MDA), oxidized LDL (oxLDL), and isoprostanes (F2-IsoP), and those associated with general oxidative conditions (ferric-reducing antioxidant power (FRAP), superoxide dismutase (SOD), glutathione (GSH)), MDA was the most lipid biomarker evaluated in the treatments and proved to be an independent factor compared with traditional markers used in the algorithms to stratify the patient's risk. Finally, this review suggests four steps to follow, aiming to include MDA in the algorithms applied to estimate CVD risk.

Effect of nanocapsules containing docosahexaenoic acid in mice with chronic inflammation.

BACKGROUND: Omega 3 fatty acids, such as docosahexaenoic acid (DHA) have been widely consumed as supplements to control chronic inflammation. Nanocapsules containing DHA (MLNC-DHA-a1) were developed and showed excellent stability. Thus, our objective was to evaluate the effect of MLNC-DHA-a1 nanocapsules on biomarkers of chronic inflammation. METHODS: Cells viability was determined by flow cytometry. The uptake of MLNC-DHA-a1 nanocapsules by macrophages and their polarization were determined. In vivo, LDLr(-,-) mice were fed a Western diet to promote chronic inflammation and were treated with MLNC-DHA-a1 nanocapsules, intravenously injected via the caudal vein once a week for 8 weeks. RESULTS: MLNC-DHA-a1 nanocapsules decreased the concentration of TNFα (p = 0.02) in RAW 264.7 cells compared to the non-treated group (NT), with no changes in IL-10 (p = 0.29). The nanocapsules also exhibited an increase in the M2 (F4/80+ CD206) phenotype (p < 0.01) in BMDM cells. In vivo, no difference in body weight was observed among the groups, suggesting that the intervention was well tolerated. However, compared to the CONT group, MLNC-DHA-a1 nanocapsules led to an increase in IL-6 (90.45 ×13.31 pg/mL), IL-1ß (2.76 ×1.34 pg/mL) and IL-10 (149.88 ×2.51 pg/mL) levels in plasma. CONCLUSION: MLNC-DHA-a1 nanocapsules showed the potential to promote in vitro macrophage polarization and were well-tolerated in vivo. However, they also increased systemic pro-inflammatory cytokines. Therefore, considering that this immune response presents a limitation for clinical trials, further studies are needed to identify the specific compound in MLNC-DHA-a1 that triggered the immune response. Addressing this issue is essential, as MLNC-DHA-a1 tissue target nanocapsules could contribute to reducing chronic inflammation.

Characterization of red wines from South America based on sensory properties and antioxidant activity.

BACKGROUND: It is widely accepted that red wines constitute one of the most important sources of dietary polyphenolic antioxidants. However, it is still not known how some variables such as variety, vintage, country of origin, and retail price are associated with the antioxidant activity and sensory profile of South American red wines. In this regard, 80 samples produced in Brazil, Chile and Argentina were assessed in relation to their sensory properties, color and in vitro antioxidant activity, and results were subjected to multivariate statistical techniques. RESULTS: Samples were grouped in clusters, characterized by high, intermediate and low in vitro antioxidant activity, sensory properties and prices. It was possible to observe that wines with high antioxidant activity were associated to high retail prices and overall perception of sensory quality. CONCLUSION: South American wines produced from Vitis vinifera such as Syrah, Malbec and Cabernet Sauvignon had higher in vitro antioxidant activity and also higher sensory quality than wines produced from Vitis labrusca. This result was independent of vintage (2002-2010), corroborating the idea that the same grape varietal, even when produced in different years, displays similar sensory characteristics and antioxidant activity.

Selenium Supplementation during Puberty and Young Adulthood Mitigates Obesity-Induced Metabolic, Cellular and Epigenetic Alterations in Male Rat Physiology.

Selenium (Se) role in obesity is not clear. In addition, information on Se's role in male physiology, specifically in obesity, is scarce. We conducted this study to evaluate the efficacy of Se supplementation, specifically during puberty until young adulthood, against obesity-induced deregulation of metabolic, cellular, and epigenetic parameters in epididymal fat and/or sperm cells in a rat model. High-fat-diet consumption by male rats during puberty and young adulthood significantly increased body weight, adipocyte size, oxidative stress, deregulated expression of genes associated with inflammation (Adiponectin, IL-6, TNF-α), adipogenesis (CEBPα), estrogen biosynthesis (CYP19) and epigenetic processes in epididymal adipose tissue (Dnmt3a), as well as altered microRNA expression vital for spermatogenesis in sperm cells (miR-15b and miR-497). On the other hand, Se supplementation significantly decreased oxidative stress and mitigated these molecular/epigenetic alterations in epididymal adipose tissue or sperm cells. Our results indicate that selenium supplementation during puberty/young adulthood could improve male physiology in the context of obesity. In addition, it suggests that Se could potentially positively affect offspring health.

Combination of natural strategies to improve the oxidative stability of echium seed oil.

Echium seed oil is an alternative source of omega 3 fatty acids but it is highly susceptible to oxidation. A combination of three natural strategies was proposed in this study aiming to improve the oxidative stability of echium oil obtained by pressing (PO) or solvent extraction (PSO), kept in the storage condition for 180 days or during the consumption for 30 days. Our results showed that the reduction of temperature was sufficient to keep the oil stable during storage for both samples. During the consumption time, the best stability was achieved by adding a mixture of antioxidants, composed of sinapic (500 ppm), ascorbic (250 ppm), and citric (150 ppm) acids, and/or 20% of high oleic sunflower oil. The combined strategies promoted a 34 to 80% reduction of peroxide value and 0 to 85% reduction of malondialdehyde concentrations in the samples, showing to be a feasible and natural alternative to improve the oxidative stability of echium oil. PRACTICAL APPLICATION: Our study successfully applied an optimized combination of simple and low-cost strategies to enhance the chemical stability of echium seed oil. As the use of echium oil expands around the world, the oil industry and final consumers may benefit from our results to increase the oil shelf-life.

Chemical characterization of Echium plantagineum seed oil obtained by three methods of extraction.

Echium seed oil has been considered an important alternative source of omega 3 fatty acids (n-3 FA) for human consumption. Considering the oxidative instability of n-3 FA richer oils, the objective of this study was to determine the chemical and sensory parameters of the oil obtained from Echium plantagineum seeds obtained by three extraction methods (hydraulic press: HYD; continuous screw press: PRESS; and solvent technique: SOLV). Stearidonic acid (C18:4, n3), the most important n-3 FA present in the oil, changed from 12.5% to 12.7%. Regarding the minor compounds, PRESS sample showed the highest concentration of gamma-tocopherol (782.24 mg/kg oil), while SOLV samples presented the highest amount of ß-sitosterol (73.46 mg/100 g) with no difference of campesterol concentration (159.56 mg/100 g) among the samples. Higher values of total phenolics (19.65 mg GAE/kg oil) and ß-carotene (34.83 mg/kg oil) were also found in the SOLV samples, suggesting the influence of hexane in the extraction of these bioactive compounds. High resolution mass spectrometry identified caffeic acid and its derivatives as the main phenolic compounds present in the echium oil. PRESS sample showed the best oxidative stability as measured by PV (0.61 mmol/kg oil) and malondialdehyde (173.13 µmol), probably due to faster time of processing compared to HYD and SOLV samples. Our data showed that the extraction method changed the chemical composition of the minor compounds in the echium oil, but these alterations did not reduce its nutritional quality or sensory acceptability. PRACTICAL APPLICATION: Echium oil represents a great potential source of omega 3 fatty acids, but there is not enough information about its oxidative stability and chemical composition, especially toward minor compounds. Our study characterizes echium oil composition obtained from three extraction methods, contributing to amplify the technical information about this important alternative oil for human consumption.

Effect of sinapic acid ester derivatives on the oxidative stability of omega-3 fatty acids rich oil-in-water emulsions.

Oil-in-water (O/W) emulsions are important delivery systems of omega-3 fatty acids (n-3 FA). We investigated the effect of sinapic acid esters concentration and chain length, the electrical charge of the emulsifier and emulsion pH on the oxidative stability of n-3 FA rich O/W emulsions. Echium oil was applied as n-3 FA source. A 24 factorial design was used to simultaneously evaluate these factors. Peroxide value, malondialdehyde, 2,4-heptadienal and 2,4-decadienal were measured in the emulsions. pH and the electrical charge of the emulsifier modulated the antioxidant effectiveness of sinapic acid esters, while concentration was not relevant. The combination of positively charged emulsifier with neutral pH provided the best oxidative stability for echium oil emulsions. Our results also suggested that the increase of length chain of sinapic acid, from C4 to C12, reduced the secondary products of oxidation, when echium oil emulsions were prepared using negatively charged emulsifier under acidic conditions.

Influence of heated, unheated whey protein isolate and its combination with modified starch on improvement of encapsulated pomegranate seed oil oxidative stability.

This study aimed at encapsulating pomegranate seed oil (PSO) by emulsification followed by spray drying using whey protein isolate (WPI) in its natural form, heated (Pickering), and combined with modified starch (WPI:Capsul®) as emulsifiers/wall materials. Emulsions were stable under different stress conditions. Pickering emulsions presented bigger droplet size (6.49-9.98 µm) when compared to WPI (1.88-4.62 µm) and WPI:Capsul® emulsions (1.68-5.62 µm). Sixteen fatty acids were identified in PSO. WPI treatment was considered the best formulation since it presented the highest fatty acid retention (68.51, 65.47, 47.27, 53.68, 52.95, and 52.28% for linoleic, oleic, punicic, α-eleostearic, catalpic, and ß-eleostearic acids after 30 days-storage, respectively) and protected the oil against volatile compound formation (heptanal, (E,E)-2,4-heptadienal, (Z)-2-heptenal, octanal, pentanal, (E)-2-hexenal, (E)-2-octenal, nonanal, (E)-2-decenal, and (E,E)-2,4-octadienal), which did not occur with free PSO. Overall, encapsulation protected PSO against oxidation over time, which may allow the development of new functional foods.

Dietary zinc deficiency or supplementation during gestation increases breast cancer susceptibility in adult female mice offspring following a J-shaped pattern and through distinct mechanisms.

Zinc is required for fetal development and is involved in key processes associated with breast carcinogenesis. We evaluated whether maternal zinc deficiency or supplementation during gestation influences female offspring susceptibility to breast cancer in adulthood. C57BL/6 mice consumed during gestation control (30 p.p.m. zinc), zinc-deficient (8 p.p.m) or zinc-supplemented (45 p.p.m.) diets. Maternal zinc supplementation increased in female mice offspring the incidence of chemically-induced mammary adenocarcinomas that were heavier, compared to control group. This was accompanied by a decreased number of terminal end buds, increased cell proliferation and apoptosis, and increased tumor suppressors p21, p53 and Rassf1, Zfp382 and Stat3 expression in mammary glands, as well as increased zinc status. Although maternal zinc deficiency did not alter the incidence of these lesions, it also induced heavier mammary adenocarcinomas, compared to control group. These effects were accompanied by a decreased number of terminal end buds, increased proto-oncogenes c-Myc and Lmo4 expression and H3K9Me3 and H4K20Me3 epigenetic marks in mammary glands of offspring, and decreased zinc status and increased levels of oxidative marker malondialdehyde. The data suggest that both maternal zinc deficiency and supplementation during gestation programmed increased breast cancer susceptibility in adult mice offspring following a J-shaped pattern through distinct mechanisms.

ß-ionone inhibits nonalcoholic fatty liver disease and its association with hepatocarcinogenesis in male Wistar rats.

Among the primary neoplasias that affect the liver, hepatocellular carcinoma (HCC) is the most frequent and the third leading cause of death related to cancer. Several risk factors predispose individuals to HCC such as nonalcoholic fatty liver disease (NAFLD), whose incidence has significantly increased worldwide. ß-ionone (ßI) isoprenoid is a known chemopreventive of hepatocarcinogenesis. However, the effects of this compound on NAFLD isolated or in association with hepatocarcinogenesis have not yet been evaluated. A high-fat emulsion administered for 6 weeks resulted in NAFLD in male rats, and oral treatment with ßI during this period significantly attenuated its development. Moreover, the presence of NAFLD potentiated hepatocarcinogenesis induced by the resistant hepatocyte (RH) model in these animals by increasing the number and percentage of the liver section area occupied by placental glutathione S-transferase (GST-P)-positive persistent preneoplastic lesions (pPNLs), that are thought to evolve into HCC. This indicates that this NAFLD/RH protocol is suitable for studies of the influence of NAFLD on the HCC development. Therefore, here we also investigated the chemopreventive effect of ßI under these two associated conditions. In this context, ßI reduced the number and percentage of the liver section area occupied by pPNLs, as well as cell proliferation and the number of oval cells, which are considered potential targets for the development of HCC. Thus, ßI presents not only a promising inhibitory effect on NAFLD isolated but also chemopreventive activity when it is associated with hepatocarcinogenesis.

Dietary glutamine supplementation increases the activity of peritoneal macrophages and hemopoiesis in early-weaned mice inoculated with Mycobacterium bovis bacillus Calmette-Guérin.

Infants who are breast-fed have been shown to have a lower incidence of certain infectious diseases compared with formula-fed infants. Glutamine is one of the most abundant amino acids found in maternal milk and it is essential for the function of immune system cells such as macrophages. The purpose of this study was to investigate the effect of glutamine supplementation on the function of peritoneal macrophages and on hemopoiesis in early-weaned mice inoculated with Mycobacterium bovis bacillus Calmette-Guérin (BCG). Mice were weaned at 14 d of age and distributed to 2 groups and fed either a glutamine-free diet (n = 16) or a glutamine-supplemented diet (+Gln) (n = 16). Both diets were isonitrogenous (with addition of a mixture of nonessential amino acids) and isocaloric. At d 21, 2 subgroups of mice (n = 16) were intraperitoneally injected with BCG and all mice were killed at d 28. Plasma, muscle and liver glutamine concentrations and muscle glutamine synthetase activity were not affected by diet or inoculation with BCG. The +Gln diet led to increased leukocyte and lymphocyte counts in the peripheral blood (P < 0.05) and granulocyte and lymphocyte counts in the bone marrow and spleen (P < 0.05). The +Gln diet increased spreading and adhesion capacities, hydrogen peroxide, nitric oxide, and tumor necrosis factor-alpha (TNFalpha) syntheses and the phagocytic and fungicidal activity of peritoneal macrophages (P < 0.05). The interaction between the +Gln diet and BCG inoculation increased the area under the curve of interleukin (IL)-1beta and TNFalpha syntheses (P < 0.05). In conclusion, the intake of glutamine increases the function of peritoneal macrophages and hemopoiesis in early-weaned and BCG-inoculated mice. These data have important implications for the design of breast milk substitutes for human infants.