Toxicological Characteristics of Bacterial Nanocellulose in an In Vivo Experiment-Part 1: The Systemic Effects.

Bacterial nanocellulose (BNC) is being considered as a potential replacement for microcrystalline cellulose as a food additive and a source of dietary fiber due to its unique properties. However, studies on the risks of consuming BNC in food are limited, and it is not yet approved for use in food in the US, EU, and Russia. AIM: This study aims to perform a toxicological and hygienic assessment of the safety of BNC in a subacute 8-week administration in rats. METHODS: BNC was administered to male Wistar rats in doses of 0, 1.0, 10.0, and 100 mg/kg body weight for 8 weeks. Various parameters such as anxiety levels, cognitive function, organ masses, blood serum and liver biochemistry, oxidative stress markers, vitamin levels, antioxidant gene expression, and liver and kidney histology were evaluated. RESULTS: Low and medium doses of BNC increased anxiety levels and liver glutathione, while high doses led to elevated LDL cholesterol, creatinine, and uric acid levels. Liver tissue showed signs of degeneration at high doses. BNC did not significantly affect vitamin levels. CONCLUSION: The adverse effects of BNC are either not dose-dependent or fall within normal physiological ranges. Any effects on rats are likely due to micronutrient deficiencies or impacts on intestinal microbiota.

Comprehensive assessment of the effectiveness of l-carnitine and transresveratrol in rats with diet-induced obesity.

OBJECTIVES: Transresveratrol (Res) and l-carnitine (l-Car) are proposed to alleviate metabolic and immune disorders and increase physical activity in obese individuals. This study aims to estimate the effect of Res and l-Car in rats with diet-induced obesity. METHODS: Male Wistar rats were fed a diet with excess fat and fructose (high-fat high-carbohydrate diet [HFCD]) supplemented with Res and l-Car at doses of 25 and 300 mg/kg of body weight, respectively, for 63 d. An assessment of grip strength, behavioral reactions, as well as biochemical, morphological, and immunological parameters, was performed. RESULTS: Res supplementation did not affect energy consumption, but l-Car increased when animals had free access to feed. Body weight gains were the highest in animals fed the HFCD, lowest in rats receiving the control balanced diet, and intermediate in animals receiving Res and l-Car. Feeding with Res and l-Car canceled the decrease in long-term memory in rats fed the HFCD, as well as reduced anxiety and increased mobility. With both supplements, bilirubin, triglycerides, and low-density lipoprotein levels in the blood plasma returned to normal values, but only l-Car increased the ratio of aspartic and alanine transaminases. In addition, l-Car lowered the levels of leptin and ghrelin and increased transforming growth factor beta 1 in the blood plasma, and consumption of Res was accompanied by a decrease in interleukin-17A and increase in interferon gamma in spleen lysates. Moreover, l-Car reduced the number of cells with lipid inclusions in the liver. CONCLUSIONS: The consumption of Res and l-Car leads to a significant reduction in dyslipidemia and inflammation with potentially favorable changes in behavioral responses.

Lipoic Acid Exacerbates Oxidative Stress and Lipid Accumulation in the Liver of Wistar Rats Fed a Hypercaloric Choline-Deficient Diet.

Non-alcoholic fatty liver disease (NAFLD) is currently estimated as the most prevalent chronic liver disease in all age groups. An increasing body of evidence obtained in experimental and clinical data indicates that oxidative stress is the most important pathogenic factor in the development of NAFLD. The study aimed to investigate the impact of α-lipoic acid (LA), widely used as an antioxidant, on the effects of a hypercaloric choline-deficient diet. Male Wistar rats were divided into three groups: control diet (C); hypercaloric choline-deficient diet (HCCD), and hypercaloric choline-deficient diet with α-lipoic acid (HCCD+LA). Supplementation of HCCD with LA for eight weeks led to a decrease in visceral adipose tissue/body weight ratio, the activity of liver glutathione peroxidase and paraoxonase-1, plasma, and liver total antioxidant activity, as well as an increase in liver/body weight ratio, liver total lipid and triglyceride content, and liver transaminase activities compared to the HCCD group without LA. In conclusion, our study shows that α-lipoic acid detains obesity development but exacerbates the severity of diet-induced oxidative stress and lipid accumulation in the liver of male Wistar rats fed a hypercaloric choline-deficient diet.

Effects of Tyrosine and Tryptophan Supplements on the Vital Indicators in Mice Differently Prone to Diet-Induced Obesity.

We studied the effects of the addition of large neutral amino acids, such as tyrosine (Tyr) and tryptophan (Trp), in mice DBA/2J and tetrahybrid mice DBCB receiving a high-fat, high-carbohydrate diet (HFCD) for 65 days. The locomotor activity, anxiety, muscle tone, mass of internal organs, liver morphology, adipokines, cytokines, and biochemical indices of animals were assessed. The Tyr supplementation potentiated increased anxiety in EPM and contributed to a muscle tone increase, a decrease in the AST/ALT ratio, and an increase in protein anabolism in both mice strains. Tyr contributed to a decrease in liver fatty degeneration and ALT reduction only in DBCB that were sensitive to the development of obesity. The addition of Trp caused an increase in muscle tone and potentiated an increase in anxiety with age in animals of both genotypes. Trp had toxic effects on the livers of mice, which was manifested in increased fatty degeneration in DBCB, edema, and the appearance of micronuclei in DBA/2J. The main identified effects of Tyr on mice are considered in the light of its modulating effect on the dopamine neurotransmitter metabolism, while for the Trp supplement, effects were presumably associated with the synthesis of its toxic metabolites by representatives of the intestinal microflora.

Detrimental effects of 6 months exposure to very low doses of a mixture of six pesticides associated with chronic vitamin deficiency on rats.

This study aimed to evaluate the long-term low-dose effects of exposure to a mixture of 6 pesticide active substances (diquat, imazamox, imazethapyr, tepraloxydin, bentazone, acifluorfen) and to elucidate if chronic vitamin deficiency can influence their toxicity. Two hundred Wistar rats were divided in 4 groups: a vitamin-sufficiency control group, a vitamin-deficiency control group, a vitamin sufficiency test group and a vitamin-deficiency test group. The test groups were treated with the aforementioned pesticides at doses 100 times lower than the corresponding NOAEL. After 6 months, ten rats from each group were sacrificed and a complete evaluation of blood and urine biochemistry, biomarkers of oxidative stress, xenobiotic detoxification enzymes and lysosomal enzymes and organ histopathology was performed. The pesticides mixture and vitamin deficiency determined an increase in alkaline phosphatase levels and urinary calcium levels, abnormal serum lipid profile, and a decrease of total blood proteins levels, red blood cells, haematocrit and haemoglobin. The combination of the two stressors up-regulated CYP1A1, CYP1A2, CYP2B1 and GST levels. This study provides a new proof for the need to move forward from single chemical testing to a more complex approach to account for the multitude of stressors that can challenge the setting of real safety levels.

Effects of Tyrosine and Tryptophan in Rats with Diet-Induced Obesity.

Amino acids tyrosine (Tyr) and tryptophan (Trp) play a significant role in the regulation of energy metabolism, locomotor activity, and eating behavior. We studied the possibility of modulating these processes in obesity by increasing the pool of Tyr and Trp in the experimental diet. As a model of obesity, we used Wistar rats fed a diet with an excess specific energy value (HFCD) for 64 days. Trp led to a normalization of the rats' body weight almost to the control level, but increased anxiety-like behavior and decreased long-term memory. The consumption of amino acids resulted in increased grip strength and impairment of short-term memory. The locomotor activity of animals decreased with age as a result of Tyr consumption, while Trp, on the contrary, prevented this. The Tyr supplementation led to the normalization of triglycerides and LDL. In the spleen cell lysates, amino acids suppressed the production of proinflammatory cytokines. The liver tissue morphology showed that the consumption of Tyr noticeably weakened the signs of fatty degeneration. The addition of Trp, on the contrary, led to an unfavorable effect, consisting of the appearance of a high number of large rounded fatty vacuoles. The data obtained indicate a more pronounced anti-inflammatory effect of Tyr as compared to Trp.

Effect of resveratrol on behavioral, biochemical, and immunological parameters of DBA/2J and tetrahybrid DBCB mice receiving diet with excess fat and fructose.

Polyphenolic biologically active substances (BAS) including resveratrol (R) can exert beneficial effects on fat accumulation, blood pressure, glycemia, insulin sensitivity, and plasma lipid profile in patients with obesity, and associated diseases. The study aimed to determine the effect of R at a dose of 25 mg/kg body weight on the DBA/2J and DBCB mice with diet-induced obesity followed by the consumption high-fat high-carbohydrate diet (HFCD). Behavioral reactions (elevated plus maze [EPM]) and muscle tone (the strength of the forepaw grip) were tested, and plasma biochemical and immunological parameters were assessed. In the repeated EPM test, anxiety increased only in DBCB mice during the second trial. In DBCB mice treated with HFCD, the muscle tone decreased with the second trial; however, this effect was not observed in the background of R consumption. R decreased the level of triglycerides, diminished the activities of alanine and asparagine aminotransferases, which were elevated upon HFCD consumption. Ghrelin level increased after R consumption in mice of both genotypes. The leptin to ghrelin ratio was reduced in DBCB mice receiving R. Consumption of R increased IL-3 and IL-10 levels in both DBA/2J and DBCB mice. IL-12p70 level increased in DBCB mice in response to R. R addition to HFCD reduced several symptoms of dyslipidemia in highly sensitive tetrahybrid mice. The results obtained indicate the importance of a personalized (depending on the genotype) approach when any R prescription, among other BAS and dietary factors, are used in diet therapy for patients with low, moderate and high-risk obesity.

Content of essential and toxic trace elements in organs of obese Wistar and Zucker leprfa rats receiving quercetin.

BACKGROUND: The levels of a number of essential and toxic trace elements in organs and tissues are affected by the disruptions in body homeostasis caused by obesity. Some of these elements may also be influenced by the consumption of biologically active substances of polyphenolic origin, which possess potent abilities to complex with transition metal ions. AIMS: The aim of this study was to determine the content of essential and toxic trace elements in Wistar outbred and hereditary obese Zucker Leprfa (Z) rats consuming a standard balanced diet or hypercaloric diet with excess fat and fructose, supplemented with quercetin or not supplemented. MATERIALS AND METHODS: Male Wistar and Z rats were fed a control AIN-93M-based semi-synthetic diet or a high-fat-high-carbohydrate diet (HFCD, with 30% fat by weight and 20% fructose provided in the drinking water). A portion of the animals in each line and diet group was administered quercetin at 50 mg/kg body weight. Essential trace elements were included in the diets as a high-purity salt mixture. After the termination of feeding on day 63, the livers, kidneys, and brains of the rats were excised and the content of 16 elements (Fe, Mg, Cu, Mn, Co, Se, Zn, Cr, Ni, Al, Cd, As, Pb, V, Cs, and Ag) was measured by inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: In the livers of the Z rats, the contents of Co, Zn, Mg, Fe, Se, and V were reduced and the content of Cr was increased compared to that of the Wistar rats. Supplementation with quercetin significantly decreased liver Fe, V, and Se content, which was more noticeable in the Wistar rats than in the Z rats. In kidneys of Z rats consuming control diet, the contents of Co, Cu, and Cs were decreased whereas those of Ni, Al, and Se were increased compared with the contents in the Wistar rats. The same trend was observed with HFCD feeding except for Cs content. Quercetin reduced kidney V content in both rat lines fed both diets, whereas it reduced Se and Cs only in the Z rats fed control diet. In the brains of the Z rats, a large increase was observed in some trace elements including Pb, Cd, Al, Cr, Ni, Fe, and V compared with the levels in the Wistar rat brains. Supplementation of the control diet with quercetin decreased Al and Ni in the brains of the Z rats. CONCLUSION: There were significant differences in the mineral content of organs between the Wistar and Z rats, with different propensities for obesity. Moreover some of these effects had no straightforward association with decreased feed consumption or hepatic fat accumulation. When introduced into the diets, quercetin affected the content of essential and toxic elements, but with ambiguous physiological significance. Thus, indicators of essential and toxic trace elements deserve to be used in the protocols of preclinical as well as clinical trials of biologically active substances and food supplements.

Effects of quercetin on the neuromotor function and behavioral responses of Wistar and Zucker rats fed a high-fat and high-carbohydrate diet.

Quercetin can affect some pathological manifestations in obesity. The mechanism underlying the presumed therapeutic effect of quercetin is probably related to the influence on the central processes regulating energy homeostasis. Thus, the purpose of this study was to examine the effect of quercetin on the neuromotor and behavioral functions in Zucker (Z) and Wistar (W) rats with genetically and/or diet-induced obesity. Rats of both strains received balanced or high fat and fructose diet (HFCD) in a 62-day experiment or the same diets supplemented with quercetin at the dose of 50 mg/kg body weight per day. The neuromotor function and behavioral responses were examined using the grip strength test, open field test, elevated plus maze test and conditioned passive avoidance response (CPAR) test. The quercetin potentiated a decrease in anxiety in W rats consumed HFCD and this effect was absent in Z rats with a defect in the leptin receptor gene. In contrast, quercetin increased locomotor activity and impaired short-term memory in the CPAR test only in Z rats with the absence of normal leptin reception. Against the background of the identified changes quercetin exerted significant effects on the lipid and nitrogen metabolism indices such as HDL cholesterol, AsAT/AlAT activities ratio, urea level as well as body and fat mass that were different in Z and W rats. The data obtained show that the effects of quercetin on behavior vary significantly between two strains of rat and consequently are mediated by processes of leptin reception.

Comparative analysis of the influence of a high-fat/high-carbohydrate diet on the level of anxiety and neuromotor and cognitive functions in Wistar and DAT-KO rats.

We compared anxiety, neuromotor, and cognitive functions in mutant rats with different allelic variants of dopamine transporter DAT knockout receiving balanced or excess in fat and fructose diet. The experiments were performed in DAT-/- homozygotes, DAT+/- heterozygotes, and DAT+/+ wild type rats. The genotype of DAT-KO rats was confirmed by restriction analysis of DAT gene compared to behavioral responses in the open field test (OF). Animals in the first groups of each strain were fed a balanced AIN93M diet; and those in the second groups with a high-fat/high-fructose diet. Neuromotor function was studied as grip strength, and behavioral responses were assessed in the elevated plus maze and conditioned passive avoidance response tests. The mass of the internal organs and white and brown fat, as well as selected lipid and nitrogen metabolism parameters in blood plasma were determined at the end of the experiment. DAT-/- had the highest specific grip strength, and showed an increase in initial exploratory activity in comparison with DAT+/- and DAT +/+. The exploratory activity was significantly reduced in the second test compared to the first one in DAT-/- and DAT+/- of first but not second group. Anxiety decreased with age in the second groups of DAT+/- and DAT+/+ (but not in DAT-/-) and was higher in DAT+/+ than in DAT+/- and DAT-/-. Excess fat and fructose resulted in the deterioration of short-term memory in DAT+/+. Lipidomic indices of blood plasma were less responsive to diet in DAT-/- and DAT-/+ in comparison to DAT+/+. The increased AsAT/AlAT activity ratio in DAT-/- compared with those in DAT+/+ suggests the activation of catabolism activity in the mutants. The consumption of excess fat and fructose significantly modified the effects produced by DAT gene allelic variants presumably due to the influence on the processes of dopamine metabolism.

Activity of xenobiotic-metabolizing enzymes in the liver of rats with multi-vitamin deficiency.

The purpose of the study was to determine how multi-vitamin deficiency affects xenobiotic-metabolizing enzyme (XME) activities in the rat liver. Vitamin levels and XME activities were studied in the livers of male Wistar rats who were fed for 4 weeks with semi-synthetic diets containing either adequate (100 % of recommended vitamin intake) levels of vitamins (control), or decreased vitamin levels (50 % or 20 % of recommended vitamin intake). The study results have shown that moderate vitamin deficiency (50 %) leads to a decrease of vitamin A levels only, and to a slight increase, as compared with the control, in the following enzyme activities: methoxyresorufin O-dealkylase (MROD) activity of CYP1 A2 - by 34 % (p < 0.05), UDP-glucuronosyl transferase - by 26 % (p < 0.05), and quinone reductase - by 55 % (p < 0.05). Profound vitamin deficiency (20 %) led to a decrease of vitamins A, E, B1, B2, and C, and enzyme activities in the liver: MROD - to 78 % of the control level (p < 0.05), 4-nitrophenol hydroxylase - to 74 % (p < 0.05), heme oxygenase-1 - to 83 % (p < 0.05), and quinone reductase - to 60 % (p < 0.05). At the same time, the UDP-glucuronosyl transferase activity and ethoxyresorufin O-dealkylase activity of CYP1A1, pentoxyresorufin O-dealkylase activity of CYP2B1/2 and 6ß-testosterone hydroxylase, as well as the total activity of glutathione transferase did not differ from the control levels. The study has demonstrated that profound multi-vitamin deficiency is associated with a decrease in the expression of CYP1A2 and CYP3A1 mRNAs to 62 % and 79 %, respectively. These data indicated that a short-term but profound multi-vitamin deficiency in rats leads to a decrease in the activities and expression of the some XME that play an important role in detoxification of xenobiotics and metabolism of drugs and antioxidant protection.