HFL Group Research Articles

Examination of the effects of excess microalgae availability on the disruption of mussel byssus secretion

The present study evaluated the effects of food availability on byssus secretion in the hard-shelled mussel Mytilus coruscus. Byssus production was evaluated, and foot tissue transcriptomes, the site of byssus production, were generated. The results showed that byssus secretion was disrupted in mussels fed high levels of microalgae (HFL, 88.8 mg/L day−1), compared to the control group (p < 0.05), and that more byssus production occurred in the HFL group during the recovery period. Byssus secretion was not affected in mussels fed low levels of microalgae (8.88 × 10−3 mg/L day−1), but the shedding of byssus filaments was significantly increased compared to the control mussels (p < 0.05). Transcriptome data of the foot tissue revealed that lysosome, FoxO signaling, and autophagy pathways involved in autophagic cell death and apoptosis were significantly affected (p < 0.05) and may explain differences in byssus growth under modified food availability in M. coruscus. High feed levels modified metabolic pathways, such as sphingolipid, tyrosine, and phenylalanine, and downregulated genes coding for mussel foot proteins, which may explain reduced byssus production. The study found that contrary to expectation increased food availability reduced mussel byssus production by altering foot metabolism and promoting autophagy of foot tissue.

Linagliptin exacerbates heart failure due to energy deficiency via downregulation of glucose utilization and absorption in a mouse model

Use of dipeptidyl peptidase-4 (DPP4) inhibitor in some clinical trials might have caused heart failure (HF), leading to increased hospitalizations. The aim of the present study was to determine whether linagliptin has any effect on chronic dilated HF, and its underlying mechanisms. Physiologic and pathologic studies were conducted on heart/muscle-specific manganese superoxide dismutase-deficient mice, which exhibited dilated cardiomyopathy, and were randomized to receive a low dose (1 mg/kg, HF-L group) or high dose (10 mg/kg, HF–H group) mixed with food, or normal food (HF group), for 8 weeks. Linagliptin increased mortality and heart/body weight ratio in mice with HF. Cardiac contractility and fibrosis worsened, whereas hepatic glycogen content and individual carbohydrate consumption decreased significantly in the HF–H group, when compared with the HF control group. Therefore, we performed a complementary experiment by supplementing glucose to the mice treated with high-dose linagliptin (HF-HG group). Adequate glucose supplementation reduced heart/body weight ratio and cardiac fibrosis, and improved cardiac contractility, without changing mortality. Following oral administration of 13C glucose, the respiratory 13C decreased in the HF–H and HF-HG groups, when compared with that in the HF group; the fecal 13C increased, suggesting that linagliptin inhibited glucose absorbance in the intestine. In addition, the expression of GLUT2, a glucose transporter was downregulated in the small intestine. Linagliptin treatment exacerbated HF, which increased mortality, cardiac function, and fibrosis. DPP4 inhibitors might boost cardiac cachexia and exacerbate HF, at least in part, through the modification of glucose utilization and absorption.

Tissue Bioengineering with Fibrin Scaffolds and Deproteinized Bone Matrix Associated or Not with the Transoperative Laser Photobiomodulation Protocol.

Extending the range of use of the heterologous fibrin biopolymer, this pre-clinical study showed a new proportionality of its components directed to the formation of scaffold with a lower density of the resulting mesh to facilitate the infiltration of bone cells, and combined with therapy by laser photobiomodulation, in order to accelerate the repair process and decrease the morphofunctional recovery time. Thus, a transoperative protocol of laser photobiomodulation (L) was evaluated in critical bone defects filled with deproteinized bovine bone particles (P) associated with heterologous fibrin biopolymer (HF). The groups were: BCL (blood clot + laser); HF; HFL; PHF (P+HF); PHFL (P+HF+L). Microtomographically, bone volume (BV) at 14 days, was higher in the PHF and PHFL groups (10.45 ± 3.31 mm3 and 9.94 ± 1.51 mm3), significantly increasing in the BCL, HFL and PHFL groups. Histologically, in all experimental groups, the defects were not reestablished either in the external cortical bone or in the epidural, occurring only in partial bone repair. At 42 days, the bone area (BA) increased in all groups, being significantly higher in the laser-treated groups. The quantification of bone collagen fibers showed that the percentage of collagen fibers in the bone tissue was similar between the groups for each experimental period, but significantly higher at 42 days (35.71 ± 6.89%) compared to 14 days (18.94 ± 6.86%). It can be concluded that the results of the present study denote potential effects of laser radiation capable of inducing functional bone regeneration, through the synergistic combination of biomaterials and the new ratio of heterologous fibrin biopolymer components (1:1:1) was able to make the resulting fibrin mesh less dense and susceptible to cellular permeability. Thus, the best fibrinogen concentration should be evaluated to find the ideal heterologous fibrin scaffold.

The DPP4 inhibitor linagliptin exacerbated heart failure due to energy deficiency via downregulation of glucose absorption and utilization in mice

Abstract Background It was reported that glucagon-like peptide-1 has cardio-protective effects, however, the occurrence of heart failure was increased with DPP4 inhibitor in some large clinical trials. Purpose The purpose of the present work was to clarify whether DPP4 inhibitor had any effects on chronic, dilated heart failure and to identify the mechanisms. Methods Heart / Muscle-specific Manganese superoxide dismutase (Mn-SOD)-deficient mice, (H/M-Sod−/−) which exert dilated cardiomyopathy were randomized to receive low dose (1 mg/kg) linagliptin, HF-L group; high dose (10 mg/kg) linagliptin, HF-H group; mixed with food, or normal food, HF group; for eight weeks. Then, evaluation was performed in cardiac function and pathophysiological changes. Result Interestingly, the mice treated with linagliptin treatment exhibited shorter life span and greater heart/body weight ratio. Echocardiographic studies showed decreased contractility in the HF-H group with compared to the HF control group. In Sirius Red staining, severer fibrosis was also observed in the linagliptin groups. HbA1c levels did not change among three groups, however, glycogen content in the liver was significantly decreased in the linagliptin groups. Similarly, indirect calorimetry showed decreased carbohydrate consumption in the HF-H group compared to the HF control group, suggesting worsening of heart failure was due to unavailability of carbohydrates as an energy source. Therefore, we fed a diet containing 33 wt% glucose to the HF-H group as complementary experiments. As a result, adequate glucose supplementation reduced cardiac fibrosis and cardiac function, while worsened life span, HbA1c levels and CHO consumption were not changed. Therefore, we hypothesized that there might be a change in the absorption of glucose in intestine or utilization ability of glucose in body another experiments were performed. 13C, which was released from mice after the oral administration of 13C glucose and measured by respiratory gas analysis, was significantly reduced in HF-H mice. The measurements the amount of 13C in feces revealed increased levels of unabsorbed glucose by linagliptin treatment in HF. In addition, GLUT2, a glucose transporter, in small intestine, was downregulated in the HF-H and the HF-HG groups. On the other hand, the cardiac uptake of Deoxy-D-glucose, 2-[1,2-3H(N)] (3H-2DG), which was injected from mice tail vein, was not different between in the HF-H group and in the HF control group. Taken together, linagliptin treatment may inhibit glucose transport in intestine in HF. Conclusions Linagliptin treatment exacerbated heart failure, resulting in shortened life span, worsened cardiac function, and fibrosis of the myocardium. DPP4 inhibitor may promote the cardiac cachexia and exacerbate heart failure at least, in part, through modification of glucose absorption and utilization. Funding Acknowledgement Type of funding sources: None.

Effect of repetitive transcranial magnetic stimulation on anxiety symptoms in patients with major depression: An analysis from the THREE-D trial.

Despite the advances in the use of repetitive transcranial magnetic stimulation (rTMS) for the treatment of major depressive disorder (MDD), there is relatively little information about its effect on comorbid anxiety symptoms. Data from a large randomized noninferiority trial comparing intermittent theta-burst stimulation (iTBS) and high-frequency (10 Hz) rTMS delivered to the left dorsolateral prefrontal cortex (HFL) were analyzed. The primary aim was assessing changes in anxiety/somatization items from the 17-item Hamilton Depression Rating Scale (HAM-D) and the Brief Symptom Inventory (BSI-A), using baseline-adjusted change with an analysis of covariance (ANCOVA), with the final scores as the outcome and baseline scores as the adjustment covariates. The analytical cohort comprised 388 participants (189 in HFL and 199 in iTBS groups). From baseline to the end of the rTMS course, the combined score from the anxiety items from the HAM-D dropped from 7.43 (SD = 2.15) to 4.24 (SD = 2.33) in the HFL group, and 7.33 (SD = 2.13) to 3.76 (SD = 2.23) in the iTBS group. The ANCOVA resulted in an effect from time (p < .0001), but not from group allocation (p = .793) or time × group interaction (p = .976). We observed mean changes in the BSI-A of -3.5 (SD = 5.4) and -3.2 (SD = 4.8), with significant effect of time (p < .0001) in the ANCOVA, but not group allocation (p = .793) or group × time interaction (.664). Our findings suggest that both 10 Hz and iTBS may yield potential reductions in anxiety symptoms when used for the treatment of MDD. Our findings warrant future research into the effects of left-sided rTMS on depressed patients struggling with concurrent anxiety symptoms.

Seed peptide lunasin ameliorates obesity-induced inflammation and regulates immune responses in C57BL/6J mice fed high-fat diet

Obesity causes immune cells to infiltrate into adipose tissues and secrete proinflammatory mediators, promoting the development of chronic diseases. The seed peptide lunasin has been reported to have several bioactivities. We aimed to investigate the immunomodulatory properties of lunasin in obese models. Female and male C57BL/6J mice were divided into three groups: low-fat diet (LF), high-fat diet (HF), and HF with an intraperitoneal injection of lunasin (HFL). In females, lunasin decreased the levels of monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-1β, and tumor necrosis factor (TNF-α) produced in peritoneal macrophages, indicating a decrease in F4/80+ macrophage infiltration, especially the CD11c + M1 phenotype. Serum leptin and tissue-oxidized lipid malondialdehyde levels were decreased in the HFL group. In males, lunasin normalized the obesity-induced increase in spleen size and splenocyte numbers. Moreover, lunasin inhibited IL-6 secretion while promoting interferon gamma (IFN-γ) and IL-2 production in the splenocytes. In vitro, lunasin increased EL-4 T-cell proliferation and IL-2 production in activated T cells under obese conditions. Thus, lunasin is a potential natural compound that promotes immunomodulation in both female and male obese mice in a sex-dependent manner. Furthermore, lunasin mediates the anti-inflammatory response and enhances the T helper type 1 cell response to obesity-related immune disorders.

Dietary n-3 but not n-6 fatty acids modulate anthropometry and fertility indices in high-fat diet fed rats: a two-generation study.

The present study assessed the modulatory potentials of dietary n-3 [α-linolenic acid (ALA, 18:3n-3, eicosapentaenoic acid (EPA, 20:5n-3 + docosahexaenoic acid (DHA) 22:6n-3), and n-6 fatty acid (LA, 18:2n-6)] on anthropometric parameters and fertility indices in high-fat-fed rats. Weanling female Wistar rats were fed with control diet (7% lard), high-fat diet (35% lard, HFL), high-fat with fish oil (21% fish oil + 14% lard, HFF), high-fat with canola oil (21% canola oil + 14% lard, HFC) and high-fat with sunflower oil (21% sunflower oil + 14% lard, HFS) for 2months, mated and continued on their diets during pregnancy. At gestation day 18-20, the intra-uterine environment was examined in representative rats, and the rest were allowed for delivering pups. The pups after lactation were subjected to mating and feeding trials as above. Growth parameters (body weight, body length (BL), abdominal circumference (AC), thoracic circumference (TC), and Lee index and fertility parameters (litter size and sex ratio) were studied. Feeding HFL diet increased BL (16%), AC (33%) and TC (21%) compared to control (p < 0.05). Adipose tissue accumulation was 11% higher in the HFL group compared to control and was lowered with n-3 fatty acid incorporation in the diet. HFL group exhibited a lower percentage of fertility, pregnancy, and delivery indices. Litter size was decreased by 20%, and litter weight was increased by 23% in HFL group compared to control with more male pups. Our study indicated that n-3 to a larger extent than n-6 fatty acids modulated high-fat induced changes in the anthropometric parameters and fertility indices.

Anti-steatotic linagliptin pleiotropic effects encompasses suppression of de novo lipogenesis and ER stress in high-fat-fed mice

AimTo investigate the effects of linagliptin treatment on hepatic energy metabolism and ER stress in high-fat-fed C57BL/6 mice. MethodsForty male C57BL/6 mice, three months of age, received a control diet (C, 10% of lipids as energy, n = 20) or high-fat diet (HF, 50% of lipids as energy, n = 20) for 10 weeks. The groups were randomly subdivided into four groups to receive linagliptin, for five weeks, at a dose of 30 mg/kg/day added to the diets: C, C-L, HF, and HF-L groups. ResultsThe HF group showed higher body mass, total and hepatic cholesterol levels and total and hepatic triacylglycerol levels than the C group, all of which were significantly diminished by linagliptin in the HF-L group. The HF group had higher hepatic steatosis than the C group, whereas linagliptin markedly reduced the hepatic steatosis (less 52%, P < 0.001). The expression of Sirt1 and Pgc1a was more significant in the HF-L group than in the HF group. Linagliptin also elicited enhanced GLP-1 concentrations and a reduction in the expression of the lipogenic genes Fas and Srebp1c. Besides, HF-L showed a reduction in the genes related to endoplasmic reticulum stress Chop, Atf4, and Gadd45 coupled with reduced apoptotic nuclei immunostaining. ConclusionLinagliptin caused a marked reduction in hepatic steatosis as a secondary effect of its glucose-lowering property. NAFLD countering involved reduced lipogenesis, increased beta-oxidation, and relief in endoplasmic reticulum stress, leading to reduced apoptosis and better preservation of the hepatic structure. Therefore, linagliptin may be used, preferably in diabetic patients, to avoid the progression of hepatic steatosis.

GLP-1 analog liraglutide-induced cardiac dysfunction due to energetic starvation in heart failure with non-diabetic dilated cardiomyopathy

BackgroundGlucagon-like peptide-1 (GLP-1) reduces cardiovascular events in diabetic patients; however, its counter-protective effects have also been suggested in patients with heart failure and the clear explanation for its mechanisms have not yet been offered.MethodsThe effects of GLP-1 analog on cardiac function and energy metabolism, especially glycemic and lipid metabolisms were elucidated using non-diabetic J2N-k hamsters which showed spontaneous dilated cardiomyopathy. J2N-k hamsters were treated with PBS (HF group), low-dose (HF-L group) or high-dose liraglutide (HF-H group).ResultsIn failing heart, GLP-1 analog exerted further deteriorated cardiac function (e.g. positive and negative dP/dt; p = 0.01 and p = 0.002, respectively) with overt fibrosis and cardiac enlargement (heart/body weight, 5.7 ± 0.2 in HF group versus 7.6 ± 0.2 in HF-H group; p = 0.02). The protein expression of cardiac muscles indicated the energy starvation status. Indirect calorimetry showed that failing hearts consumed higher energy and carbohydrate than normal hearts; moreover, this tendency was augmented by GLP-1 analog administration. Upon 10% glucose solution loading with GLP-1 analog administration (HF-H-G group) as complementary experiments, the cardiac function and fibrosis significantly ameliorated, whereas carbohydrate utilization augmented further and lipid utilization reduced more. The prognosis of HF-H-G group also significantly improved (p = 0.025).ConclusionsGlucagon-like peptide-1 analog caused the relative but desperate shortage of glycemic energy source for the failing cardiac muscles and it may restrict ATP synthesis, resulting in cardiac function deterioration. Therefore, appropriate energy supply and amount of carbohydrate intake should be carefully considered when administrating incretin-related drugs to patients with heart failure.

Delta-6-desaturase (FADS2) inhibition and omega-3 fatty acids in skeletal muscle protein turnover

Polyunsaturated fatty acids (PUFAs) are essential dietary components. They are not only used for energy, but also act as signaling molecules. The delta-6 desaturase (D6D) enzyme, encoded by the FADS2 gene, is one of two rate limiting enzymes that convert the PUFA precursors – α-linolenic (n-3) and linoleic acid (n-6) to their respective metabolites. Alterations in the D6D enzyme activity alters fatty acid profiles and are associated with metabolic and inflammatory diseases including cardiovascular disease and type 2 diabetes. Omega-3 PUFAs, specifically its constituent fatty acids DHA and EPA, are known for their anti-inflammatory ability and are also beneficial in the prevention of skeletal muscle wasting, however the mechanism for muscle preservation is not well understood. Moreover, little is known of the effects of altering the n-6/n-3 ratio in the context of a high-fat diet, which is known to downregulate protein synthesis. Twenty C57BL6 male mice were fed a high-fat lard (HFL, 45% fat (mostly lard), 35% carbohydrate and 20% protein, n-6:n-3 PUFA, 13:1) diet for 6 weeks. Mice were then divided into 4 groups (n = 5 per group): HFL– , high-fat oil– (HFO, 45% fat (mostly Menhaden oil), 35% carbohydrate and 20% protein, n-6:n-3 PUFA, 1:3), HFL+ (HFL diet plus an orally administered FADS2 inhibitor, 100 mg/kg/day), and HFO+ (HFO diet plus an orally administered FADS2 inhibitor, 100 mg/kg/day). After 2 weeks on their respective diets and treatments, animals were sacrificed and gastrocnemius muscle harvested. Protein turnover signaling were analyzed via Western Blot. 4-EBP1 and ribosomal protein S6 expression were measured. A two-way ANOVA revealed no significant change in the phosphorylation of both 4EBP-1 and ribosomal protein S6 with diet or inhibitor. There was a significant reduction in STAT3 phosphorylation with the inhibition of FADS2 (p = 0.03). Additionally, we measured markers of protein degradation through levels of FOXO phosphorylation, ubiquitin, and LC3B expression; there was a trend towards increased phosphorylation of FOXO (p = 0.08) and ubiquitinated proteins (p = 0.05) with FADS2 inhibition. LC3B expression, a marker of autophagy, was significantly higher in the HFL plus FADS2 inhibition group from all other comparisons. Lastly, we analyzed activation of mitochondrial biogenesis which is closely linked with protein synthesis through PGC1-α and Cytochrome-C expression, however no significant differences were associated with either marker across all groups. Collectively, these data suggest that the protective effects of muscle mass by omega-3 fatty acids are from inhibition of protein degradation. Our aim was to determine the role of PUFA metabolites, DHA and EPA, in skeletal muscle protein turnover and assess the effects of n-3s independently. We observed that by inhibiting the FADS2 enzyme, the protective effect of n-3s on protein synthesis and proliferation was lost; concomitantly, protein degradation was increased with FADS2 inhibition regardless of diet.

Maoberry (Antidesma bunius) Improves Glucose Metabolism, Triglyceride Levels, and Splenic Lesions in High-Fat Diet-Induced Hypercholesterolemic Rats.

Excessive consumption of a high-fat diet (HFD) is associated with hypercholesterolemia and cardiovascular disease (CVD). Dark purple maoberry (Antidesma bunius) fruit is a very good source of antioxidants. We investigated the effects of maoberry on immune function, lipid profiles, and oxidative stress in HFD-induced hypercholesterolemia. Seventy-two male Sprague Dawley rats were divided into the normal group fed with standard diet (ND); HFD groups (HF); and low, medium, and high dose of maoberry extract groups and a simvastatin group (HF-L, HF-M, HF-H, and HF-S, respectively). Maoberry groups were given maoberry extract at concentrations of 0.38, 0.76, and 1.52 g/kg per day. At the same time, HF-S groups were administered simvastatin 10 mg/kg per day. After 12 weeks of maoberry treatment, significant reductions in body weight and triglyceride levels were observed in HF-L, HF-M, and HF-H groups in comparison with HF groups (P < .05). Obvious negative changes in spleen histology were found in HF groups, but not in maoberry-treated groups. Modest, but not significant, improvements were observed in other lipid profiles, immune cells in peripheral blood, oxidative stress, and antioxidant capacity after maoberry supplementation. In summary, these findings suggest that maoberry was helpful in reducing atherogenic risk factors such as lipid profiles, especially triglyceride, inflammation, oxidative stress related to CVD, and lesions in spleen histopathology.

The effect of daily milk production on the milk composition and energy management indicators in Holstein–Friesian and Simmental cows

The present study investigated the effect of daily milk production on the content of crucial milk components, body condition, and the level of energy management parameters in the blood and milk in Holstein?Friesian (HF) and Simmental (SIM) cows. Within each breed, the animals were divided into two groups: the HF-L group was HF cows with daily milk production of ≤31 kg and the HF-H group was those with daily milk production of >31 kg. The SIM-L was SIM cows with daily milk production of ≤27 kg and the SIM-H group was those with daily milk production of >27 kg. The intensity of reserve fat accumulation was assessed by using body condition scoring. Samples of milk were taken every 30 days of lactation and the content of total protein, casein and ?-casein, fat, lactose, dry matter, and urea were determined. Blood samples were taken on days 10, 40, 70, 100 and 130 of lactation to determine the content of glucose and beta-hydroxybutyrate. The obtained results suggest that, in spite of the lower milk production, the Simmental cows had the more favorable energetic homeostasis of the organism. The values of casein and κ-casein indicate better suitability of the raw material obtained from Simmental cows for cheese processing. From a practical point of view, higher Holstein?Friesian cows? average daily yield and a relatively small difference in the content of the total protein compared to Simmental cows? milk suggest its use primarily as consumption milk.

Dietary n-3 but not n-6 fatty acids down-regulate maternal dyslipidemia induced inflammation: A three-generation study in rats

This study addresses the effects of n-3 and n-6 fatty acids in maternal dyslipidemia induced inflammation over three generation in rats. The detailed protocol for animal feeding and mating is described in the methodology. Placenta and fetal liver were isolated on the eighteenth day of gestation and delivered pups after lactation were kept on their maternal diets. Compared to control and experimental groups, high-fat fed rats (HFL) had a higher level of cytokines and eicosanoids in serum (p < 0.05). Liver and uterine expression of cPLA-2, Cox-2, 5-Lox, EP-1, BLT-1, and ICAM-1 were higher (p < 0.05) in HFL group. NF-kB and Nrf-2 levels in placenta and fetal liver were beneficially modulated by n-3 but not n-6 fatty acids. Offspring of dyslipidemic mothers’ exhibit amplified inflammatory markers when continued on diets of their mothers. Incorporation of n-3 but not n-6 fatty acids down-regulated maternal dyslipidemia induced inflammatory markers.

A study on the Effects of Lactobacillus fermantum-fermented Milk Consumption on Lipid Patterns of Rats Fed with a High Fat Diet

Introduction: Hypercholesterolemia is a risk factor for cardiovascular diseases. Several studies have shown that some lactobacillus strains can reduce cholesterol. The purpose of this study was to evaluate the effects of Lactobacillus fermentum isolated from the traditional yogurt on body weight and lipid trends in rats fed with a high-fat diet. Materials & methods: Forty male rats (200±20 gr) were randomly divided into four groups. Rats in the first group received normal diet (N), the second group consumed normal diet with fermented milk containing L. fermentum (N-L), the third group received a high-fat diet to induce hypercholesterolemia (HF), and the fourth group were fed in a high-fat diet with fermented milk containing L. fermentum (HF-L). After 8 weeks, body weights and lipid metabolisms were measured. Findings: In the rats fed with a high-fat diet of fermented milk containing L. fermentum (HF-L), serum total cholesterol, LDL-C, and triglycerides levels were significantly lower than that of the group fed with a high-fat diet without probiotic supplementation (HF), whereas the serum HDL-C level significantly increased (p 0.05). However, mean levels of serum cholesterol, LDL-C, triglycerides, and HDL-C were not markedly different among the N groups (N, N-L). Also, the gain in body weight of the HF group was significantly heavier than that of the HF-L group (p 0.01). Discussion & conclusions: This study suggests that consumption of milk fermented by L. fermentum improves serum lipid trends in rats by lowering serum total cholesterol, triglycerides, LDL-C levels, as well as by increasing HDL-C level. It also plays a role in the prevention of obesity induced by a high-fat diet.

Effects of Fish‐Oils on Markers of Hepatic Steatosis and Cholesterol Homeostasis in C57BL/6 Mice

Non‐alcoholic fatty liver disease (NAFLD), characterized by hepatic lipid accumulation exceeding 7% of total liver mass, is the most prevalent liver disease in America. Recent evidence proposes that altered cholesterol metabolism during NAFLD may drive the liver disease progression to non‐alcoholic steatohepatitis (NASH). In this cholesterol‐focal model of NASH, imbalances in cholesterol homeostasis promotes genes that stimulate de novo lipogenesis and chronic inflammation. Emerging evidence; however, has shown diets rich in fish‐oils have strong inhibitory effects on hepatic triglyceride accumulation and this may be due in part to restoring cholesterol homeostasis as determined by the total cholesterol to free cholesterol ratio. Therefore, the purpose of this study was to determine the effects of fish‐oils on markers of hepatic steatosis and cholesterol homeostasis in C57BL/6 mice. In our study, male C57BL/6 mice were randomly assigned to four dietary groups based on percentage and type of dietary fat for 32 weeks: 10% lard (LFL), 10% fish‐oil (LFFO), 41% lard (HFL), and 41% fish‐oil (HFFO). A two‐way factorial ANOVA was performed to identify significant (p&lt;0.05) differences between fat amount (low‐fat vs. high‐fat) and fat type (lard vs. fish‐oils) on markers of hepatic steatosis, cholesterol homeostasis, and dyslipidemia. At the end of 32 weeks (Table 1), the HFL group was significantly heavier (p&lt;0.001) than the other diet groups, despite for no significant difference between groups for average calories consumed per week. When compared to HFFO, mice in the HFL group showed higher hepatic total lipid percentage and triglyceride concentration by 89% (p&lt;0.001) and 1.4‐fold (p&lt;0.001), respectively. No significant difference was observed between LFL and LFFO for body mass, hepatic lipid percentage, hepatic cholesterol, net glucose AUC, plasma insulin, plasma glucose, or HOMA‐IR. HFFO mice showed a 3‐fold (p&lt;0.001) lower hepatic total cholesterol and 5‐fold (p&lt;0.001) lower hepatic free cholesterol, as well as a 43% (p&lt;0.001) lower free cholesterol:total cholesterol ratio when compared to HFL mice. This may be due to improved insulin sensitivity bolstered by fish‐oils, as HFFO mice showed 2.2‐fold (p&lt;0.001) lower HOMA‐IR, 30% (p&lt;0.001) lower net glucose AUC, 1.3‐fold (p&lt;0.001) lower plasma insulin, and 43% (p&lt;0.001) lower plasma glucose when compared to HFL mice. Based on previous studies and our data taken together, we believe that the improvements in hepatic steatosis in mice consuming a diet high in fish‐oils was due to collective improvements in insulin sensitivity and a lower free cholesterol:total cholesterol ratio. In summary, fish‐oils improved hepatic lipid profiles that were associated with cholesterol homeostasis and insulin sensitivity.Support or Funding InformationR21 NS090282‐01This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

PP.12.03] EFFECTS OF LOSARTAN ON PROSTANOIDS RELEASE IN TWO MODELS OF METABOLIC ALTERATION IN THE RAT

Objective: Fructose-overload and high-fat diet in rats are experimental models that resemble human metabolic syndrome. This multifactorial alteration is associated to hypertension. The systemic renin-angiotensin system plays an important role in the regulation of vascular tone. Prostanoids (PR) are endogenous substances derived from arachidonic acid via the cyclooxygenases with vasoactive effects. The aim of this study was to analyze the effects of losartan (L, angiotensin 1 receptor antagonist) on prostanoids (PR) release by the mesenteric vascular bed (MVB) in both models of dietary alteration. Design and method: Six groups (n = 6) of male Sprague-Dawley rats were studied during 9 weeks: Control (C), standard diet (SD) and tap water to drink; fructose-overloaded (F), SD and fructose solution (10% W/V) to drink; HF diet (HF), 50% (w/w) bovine fat added to SD and tap water; C + L (CL), SD + 30 mg/Kg/day L in the drinking water; F + L (FL) 30 mg/Kg/day L in the fructose solution; and HF + L (HFL) 30 mg/Kg/day L in the drinking water. MVBs were removed and incubated and the released PRs measured by HPLC. Results: F and HF increased systolic blood pressure (SBP, mmHg, 133 ± 3, 145 ± 5 vs. C: 116 ± 2, p < 0.01 and p < 0.001). In the FL and HFL groups, L decreased SBP (102 ± 5 vs. F, 111 ± 3 vs. HF, p < 0.001 and p < 0.01). F diminished prostaglandin (PG) 6-ketoF1alpha, stable metabolite of the vasodilator prostacyclin (PGI2), (ng PR/mg tissue, 58 ± 11 vs. C: 98 ± 8, p < 0.05); and PGE2 (ng/mg, 35 ± 7 vs. C: 87 ± 6, p < 0.01). HF diet increased thromboxane (TX) B2, stable metabolite of the vasoconstrictor TXA2, (ng PR/mg tissue, 111 ± 5 vs. C: 64 ± 7, p < 0.001); and PGF2alpha (150 ± 10 vs. C: 83 ± 8, p < 0.01). In the FL group, L increased PG6-ketoF1alpha (117 ± 10 vs. F, p < 0.05) and PGE2 (104 ± 11 vs. F, p < 0.01). In the HFL group, L decreased TXB2 (66 ± 7 vs. HF, p < 0.01); and PGF2alpha (90 ± 7 vs. HF, p < 0.05). Conclusions: In conclusion, one of the possible mechanisms by which L reduced blood pressure in these models could be an improvement of endothelial function through a regulation of PR release which produced a decrease in vasoconstriction.

Improved growth performance, food efficiency, and lysine availability in growing rats fed with lysine-biofortified rice

Rice is an excellent source of protein, and has an adequate balance of amino acids with the exception of the essential amino acid lysine. By using a combined enhancement of lysine synthesis and suppression of its catabolism, we had produced two transgenic rice lines HFL1 and HFL2 (High Free Lysine) containing high concentration of free lysine. In this study, a 70-day rat feeding study was conducted to assess the nutritional value of two transgenic lines as compared with either their wild type (WT) or the WT rice supplemented with different concentrations of L-lysine. The results revealed that animal performance, including body weight, food intake, and food efficiency, was greater in the HFL groups than in the WT group. Moreover, the HFL diets had increased protein apparent digestibility, protein efficiency ratio, and lysine availability than the WT diet. Based on the linear relationship between dietary L-lysine concentrations and animal performance, it indicated that the biological indexes of the HFL groups were similar or better than that of the WT20 group, which was supplemented with L-lysine concentrations similar to those present in the HFL diets. Therefore, lysine-biofortified rice contributed to improved growth performance, food efficiency, and lysine availability in growing rats.

Impact of Maternal Obesity and High Fat Diet during Pregnancy and Lactation on Offspring Body Weight, Immune Function, and Resistance to Influenza Infection

Studies have suggested that obesity is associated with immune dysfunction and higher risk of infection. Maternal obesity is thought to affect body weight which may in turn alter immune function in offspring. However, the relative contribution of maternal factors (maternal obesity versus high fat diet during pregnancy and lactation) to the changes in body weight, immune function, and response to infection in the offspring is not well‐understood. To address this, we fed outbred 1‐mo old female CD‐1 mice a high fat (HF, 45%) diet or a low fat (LF, 10%) control diet to induce obesity. After 5‐mo on the respective diets, mice fed the high fat diet had considerable variation in weight gain. Thus, based on body weight, we further divided the mice in the HF group into two sub‐groups: obesity‐prone (high fat obese, HF‐O) and obesity‐resistant (high fat lean, HF‐L) groups. This provided us with the opportunity to separate the impact of obesity from that of high fat diet during pregnancy. Dams in all 3 groups (LF, HF‐O, HF‐L) were mated with 3‐mo old male CD1 mice fed a standard chow diet. All dams continued to be fed their respective diet during pregnancy and lactation. The offspring born to LF, HF‐L, and HF‐O dams were fed standard chow and monitored for their body weight. At 10‐mo of age, they were infected with influenza virus (A/Puerto Rico/8/34) and then monitored daily for symptoms until day 8 post‐infection when they were euthanized and tissues collected for analysis. Results showed that while offspring in all groups exhibited post‐infection weight loss, offspring of HF‐L group had the most weight loss. Further, both male and female offspring born to HF‐L dams had significantly more severe lung pathology compared to those born to HF‐O dams. In addition, female offspring born to HF‐O dams had significantly higher CD3 or CD3/CD28‐stimulated lymphocyte proliferation than those born to LF dams, and also had significantly lower % cytotoxic T cells (CD8+ cells) than those born to LF dams and HF‐L dams. Percentage of total T cells (CD3+), helper T cells (CD4+), and regulatory T cells (CD4+CD25+) in spleen were significantly lower in the male offspring of HF‐O dams compared with the male offspring of HF‐L dams. Taken together our results showed that maternal obesity, independent of high fat diet, influences immune response and resistance to influenza infection. Contrary to our expectation, offspring born to obese mothers had less pathology and weight loss and better T cell function following influenza infection. While the underlying mechanisms of varied resistance to influenza infection are not completely understood, our results suggest that they might be, at least in part, related to differences in T cell profile and function.Support or Funding InformationSupported by the USDA under contract #58‐1950‐4‐003

Dietary omega-3 but not omega-6 fatty acids down-regulate maternal dyslipidemia induced oxidative stress: A three generation study in rats

BackgroundMaternal nutrition modulates fetal metabolic programming and development later. Maternal dyslipidemia effects on oxidative stress (OS) in offsprings and its modulation by dietary fatty acids over generations remains to be elucidated. The objective of present study was to assess the long-term (three generations) effect of omega-3 fatty acids on OS under dyslipidemia. MethodsWeanling female Wistar rats were fed with control diet (7% lard), high fat diet (35% lard, HFL), high fat with fish oil (21% fish oil + 14% lard, HFF), high fat with canola oil (21% canola oil + 14% lard, HFC) and high fat with sunflower oil (21% sunflower oil + 14% lard, HFS). Following 60 days feeding, the female rats were mated with sexually matured males (fed normal chow diet) and continued with the above diet regimen during pregnancy and lactation. The pups after lactation were continued with their maternal diet for 60 days and subjected to mating and feeding trial as above for two generations. Serum lipid profiles, OS markers (lipid peroxidation, nitric oxide release and protein carbonyl) and antioxidant defence enzymes (catalase, SOD, glutathione peroxidase and glutathione transferase) were assessed in serum, liver and uterus of rats fed on experimental and control diets for three generations. ResultsFeeding HFL diet increased blood lipids, OS and lowered the antioxidant enzymes activity in serum, liver and uterus (p < 0.05). The reduction in the antioxidant enzymes in HFL group were higher in third followed by second generation compared to first generation (p < 0.05). Omega-3 fatty acids prevented the dyslipidemia induced loss of antioxidant enzyme activities in serum, liver and uterus. ConclusionsOur data show for the first time that offsprings born to dyslipidemic mothers’ exhibit diminished enzymatic antioxidant defence and its progressive reduction in future generation, and dietary omega-3 fatty acids restore the enzymatic antioxidant defence in offsprings and suppress the markers of OS.

A high-fish-oil diet prevents adiposity and modulates white adipose tissue inflammation pathways in mice

Fish oil improves obesity and its comorbidities, but its mechanisms of action remain unknown. We evaluate the effects of a diet rich in fish oil in white adipose tissue (WAT) inflammation pathways, renin–angiotensin system (RAS) and mitogen-activated protein kinases (MAPKs). To achieve our aims, four groups of male C57BL/6 mice were fed different diets: standard chow diet (SC; 10% energy from fat), SC+fish oil diet (SC-FO; 10% energy from fat), high-fat lard diet (HF-L; 50% energy from lard) and HF fish oil diet (HF-FO; 50% energy from fish oil). We evaluated body mass, epididymal fat pad mass, food intake and glucose tolerance. In WAT, we assessed adipocyte hypertrophy, monocyte chemotactic protein-1 immunofluorescence, and gene and protein expression of insulin signaling, inflammation, MAPKs, RAS, peroxisome proliferator-activated receptors (PPARs) and AMP-activated protein kinase (AMPK). In relation to the results, the HF-L group, as expected, showed elevated body mass and adiposity, glucose intolerance and hypertrophied adipocytes. In WAT, we found a defect in insulin signaling, infiltration of macrophages and inflammatory markers with the associated activation of MAPKs and local RAS. On the contrary, the HF-FO group did not present increased body mass, adiposity or glucose intolerance. In this group, insulin signaling, macrophage infiltration and inflammation were reduced in WAT in comparison with the HF-L group. We also observed decreases of MAPKs and local RAS and elevation of PPAR and AMPK. In summary, fish oil activates PPAR (the three isoforms) and AMPK, decreases WAT insulin resistance and inflammation, and inhibits MAPK and RAS pathways activation.