Non-alcoholic fatty liver disease (NAFLD) represents the most common chronic liver disease in industrialized countries. NAFLD progresses through the inflammatory phase of non-alcoholic steatohepatitis (NASH) to fibrosis and cirrhosis, with some cases developing liver failure or hepatocellular carcinoma (HCC). Liver biopsy remains the gold standard approach to a definitive diagnosis of NAFLD and the distinction between simple steatosis and NASH. The pathogenesis of NASH is still not clear. Several theories have been proposed ranging from the "Two Hit Theory" to the "Multiple Hit Theory". However, the general consensus is that the gut microbiota, oxidative stress, and mitochondrial damage play key roles in the pathogenesis of NASH. The interaction between the gut epithelia and some commensal bacteria induces the rapid generation of reactive oxygen species (ROS). The main goal of any therapy addressing NASH is to reverse or prevent progression to liver fibrosis/cirrhosis. This problem represents the first "Achilles' heel" of the new molecules being evaluated in most ongoing clinical trials. The second is the inability of these molecules to reach the mitochondria, the primary sites of energy production and ROS generation. Recently, a variety of non-pharmacological and pharmacological treatment approaches for NASH have been evaluated including vitamin E, the thiazolidinediones, and novel molecules related to NASH pathogenesis (including obeticholic acid and elafibranor). Recently, a new isoform of human manganese superoxide dismutase (MnSOD) was isolated and obtained in a synthetic recombinant form designated rMnSOD. This protein has been shown to be a powerful antioxidant capable of mediating ROS dismutation, penetrating biological barriers via its uncleaved leader peptide, and reducing portal hypertension and fibrosis in rats affected by liver cirrhosis. Based on these distinctive characteristics, it can be hypothesized that this novel recombinant protein (rMnSOD) potentially represents a new and highly efficient adjuvant therapy to counteract the progression from NASH to HCC.

ObjectiveLutein, an essential micronutrient that plays a critical role in promoting immune health, is being developed commercially (as Lutea® 20) using a novel manufacturing process. Lutea® 20 features reduced lutein crystal size (>99%<0.5 microns) compared to an existing lutein‐containing product, Existing Lutein Product (ELP) (64%<0.5 microns), along with enhanced stability without the requirement for including protective antioxidants. The objective of this study was to compare the oral bioavailability of lutein derived from Lutea® 20 (manufactured using novel technology) to that of lutein derived from ELP (manufactured using traditional technology).MethodsThe study was performed in 12 healthy Beagle dogs [pre‐study mean body weight (bw) of 14.3 ± 2.6 kg)] using a cross‐over, 4 × 4 Latin square design. Each animal was randomly assigned to one sequence group, and 1 of 4 treatments administered by oral gavage during each period, i.e., Lutea® 20 at 2.5 and 5 mg/kg bw and ELP at 2.5 and 5 mg/kg bw. All animals received all the treatments in sequence, with a between‐period wash‐out of 13–15 days. For each period, blood samples were collected from each animal at −24 h (pre‐administration sample), and at 2 h, 4 h, 6 h, 9 h, 12 h, 18 h, 24 h, 36 h, and 48 h post‐dosing. Lutein concentration in plasma was determined using a novel, sensitive (theoretical limit of detection = 315 pg/ml; lower limit of quantitation = 2.5 ng/ml; upper limit of quantitation = 125 ng/ml), and precise [interday % coefficients of variation: 2.7% (2.5 ng/ml), 5.2% (12.5 ng/ml), 7.2% (125 ng/ml)] high performance liquid chromatographic method with ultraviolet‐visible detection, validated as specified by the European Agency for the Evaluation of Medicinal Products and the European Medicines Agency. Maximum plasma concentration (Cmax) and the area under the curve (AUC)0–48h were calculated for each animal in each period. Data were analyzed by analysis of variance followed Tukey's‐b post‐hoc test to determine differences in the Cmax and AUC0–48h among groups.ResultsFor both doses of each formulation, maximum plasma lutein concentrations occurred at 4–6 h and gradually returned to baseline by 24 h. For each dose, the Cmax (89, 112, 60, 79 ng/ml for 2.5 and 5 mg/kg Lutea, 2.5 and 5 mg/kg ELP, respectively) and the AUC0–48h (1390, 1874, 986, 1324 ng • h/ml for 2.5 and 5 mg/kg Lutea® 20, 2.5 and 5 mg/kg ELP, respectively) for lutein were approximately 40–50% greater in the Lutea® 20 formulation compared to the existing formulation (P < 0.05, Lutea® 20 vs ELP, for each parameter at each dose).ConclusionLutein (Lutea® 20), produced using a novel manufacturing technology, possesses superior oral bioavailability compared to an existing lutein‐containing product (ELP), and this observed superiority was likely due to the smaller crystal size of lutein in Lutea® 20 compared to lutein in ELP.Support or Funding InformationNovus International, St. Charles, MO USA

Objectives: This study was performed to evaluate the potential efficacy of natural eggshell membrane (NEM) in collagen-induced arthritic rats, a well-established rodent model of inflammation and rheumatoid arthritis. Methods: Rats with developing type II collagen-induced arthritis (CIA) were treated once daily by oral gavage on study days - 14 to 17 with vehicle or NEM (52 mg/kg body weight). Rats were euthanized on study day 17. Efficacy was assessed by daily ankle caliper measurements, ankle diameter expressed as area under the curve (AUCd0-17), and histopathologic evaluation of ankles and knees. Serum biomarkers of cartilage function and inflammation [ collagen type II C-telopeptide (CTXII), cartilage oligomeric matrix protein (COMP), and alpha-2-macroglobulin (A2M)] were measured by ELISA. Results: Treatment with NEM resulted in significant beneficial effects on the daily ankle diameter measurements and ankle diameter AUC. Ankle and knee histopathology scores were significantly reduced (36% and 43% reduction of summed individual histopathology scores for ankle and knee, respectively; p< 0.05) toward normal for rats given NEM compared to vehicle controls. The percent reduction of serum CTXII, COMP, and A2M in NEM-treated rats ranged from 30% to 72% (p< 0.05). Conclusions: NEM significantly improved multiple aspects of inflammatory arthritis including inflammation, pannus, cartilage damage, bone resorption, and periosteal bone formation. This study provides further support for the use of CTXII, COMP, and A2M as relevant biomarkers that were responsive to NEM.

The composition of an individual's microbiome significantly impacts their health and risk of developing intestinal and systemic diseases. The oral microbiome is a major contributor to our oral health, which in turn is intimately linked to systemic health, the ability to fend off disease, and our overall quality of life. The use of selected, orally-targeted probiotics now offers a highly complementary approach that promises to deliver the next generation of oral and throat healthcare. This article discusses recent published studies (2012-present) that highlight the clinical evaluation of selected probiotics for halitosis, throat infections, and dental health.

The authors van den Born et al. (1) have written a timely Perspective in this issue of Diabetes . Both type 1 and type 2 diabetes have reached epidemic proportions throughout the world, afflicting over 400 million people. Moreover, the number of individuals that will develop diabetes is predicted to rise (2). Both individuals with type 1 and type 2 diabetes are at a significantly greater risk for developing microvascular and macrovascular diseases. People with diabetes who cannot maintain adequate glycemic control (such as the failure to reach the recommended target level of HbA1c <7%) are predisposed to develop neuropathy, retinopathy, nephropathy, cardiovascular disease, cerebrovascular disease, and premature death. In response to the enormity of this medical problem, there have been major initiatives on the part of global health organizations, national diabetes associations, and primary caregivers to educate patients about the benefits of appropriate nutrition and physical activity. For individuals with diabetes who have insufficient appropriate nutrition and physical activity, an increasing number of oral and injectable interventions are available to improve glycemic control (3,4). For many patients, however, the current forms of therapy now used for treating both types of diabetes are inadequate. Thus, there clearly remains a large area of unmet therapeutic need for novel pharmacological interventions that target the major complications of diabetes. Such therapies need to be identified and developed with greater efficiency by exploiting innovative molecular targets. In the current Perspective, van den Born et al. (1) present interesting data suggesting that the modulation of one or more of the three major gasotransmitters (nitric oxide [NO], carbon monoxide [CO], and hydrogen sulfide [H2S]) could eventually offer a novel therapeutic option(s) targeting the vascular complications …

The goal of this study was to compare the antagonism of elevated dietary Cu (250 mg/kg) from CuSO4 on three different Zn sources (ZnSO4 · H2O; [Zn bis(−2-hydroxy-4-(methylthio)butanoic acid)], Zn(HMTBa)2, a chelated Zn methionine hydroxy analogue; and Zn-Methionine), as measured using multiple indices of animal performance in ROSS 308 broilers.

Purpose: To determine the efficacy of an equine joint supplement STEADFAST ® and/or its active components (Natural Eggshell Membrane

Animal diets often contain antagonists that reduce Zn bioavailability, thereby creating a deficiency. The bioavailability of Zn chelated to 2-hydroxy-4-(methylthio)butanoic acid (chelated Zn) was compared with Zn sulfate in broiler chicks in two dietary conditions: a Zn-deficient corn-soybean meal (C-SBM) diet and a Zn-deficient C-SBM diet containing elevated Ca and P as antagonists. In experiment 1, chicks were fed a common diet (19 mg Zn/kg diet) through day 8, and then a Zn-unsupplemented (21 mg Zn/kg diet; 0.82% Ca, 0.47% available P) C-SBM basal or the basal supplemented with 5, 10, 15, 20, or 30 mg Zn/kg diet as chelated Zn or Zn sulfate for 6 days. Common-intercept, multiple linear regression slope-ratio analysis indicated significantly greater Zn bioavailability for chelated Zn relative to Zn sulfate as judged by: total tibia Zn (µg) (161%; =0.001); tibia zinc concentration (µg/g; 165%, =0.0009); and small intestinal metallothionein mRNA expression (248%; =0.009). In experiment 2, chicks were fed a Zn-deficient (24 mg Zn/kg diet) common diet, then fed a C-SBM basal elevated in Ca and P (27 mg Zn/kg diet; 1.2% Ca, 1% available P), or the basal supplemented with 15 or 30 mg Zn/kg diet as Zn sulfate or 7.5, 15, or 30 mg Zn/kg diet as chelated Zn. Zn sulfate was antagonized to a greater extent than Zn chelate. Bioavailability of chelated Zn relative to Zn sulfate was 441% ( =0.0063; µg total tibia Zn), 307% ( =0.0066; µg/g tibia Zn), and 426% ( =0.0041; metallothionein). Thus, feeding chelated Zn offers advantages over inorganic Zn, especially in diets containing high levels of Ca and P, which is a common occurrence in pet food and in livestock diets. antagonism, broilers, chelate, metallothionein, minerals

OBJECTIVE:Lipoic acid (LA) is a widely used nutritional supplement and is sometimes used as an adjuvant treatment for diabetic neuropathy and other conditions. Insulin autoimmune syndrome (IAS, Hirata disease) is a rare cause of spontaneous hypoglycaemia, extremely high serum insulin levels and high titres of autoantibodies against endogenous insulin despite no prior exposure to exogenous insulin. In Japanese individuals, IAS is associated with the human leucocyte antigen (HLA) HLA-DRB1*04:06 allele and often occurs upon exposure to sulphhydryl-containing compounds including LA. Only one case has been reported in Caucasians. We now report six Caucasian patients taking LA with IAS and describe a unique HLA subtype in these patients.RESEARCH DESIGN AND METHODS:Six Caucasian patients (M = 3; F = 3), median age 63 years, presented with spontaneous episodes of fasting and postabsorptive hypoglycaemia associated with mainly neuroglycopenic symptoms. No patient was treated with insulin or had an insulinoma. Hypoglycaemic symptoms appeared 30 and 120 days after taking lipoic acid (LA; 600 mg/day). Case histories and standard laboratory analyses were utilized.RESULTS:Discontinuation of LA resulted in a reduction in hypoglycaemic episodes. All patients were treated with oral or iv glucose and prednisone (12.5-25 mg/day). HLA analysis revealed the HLA-DRB1*04:03 allele in five patients, while the HLA-DRB1*04:06 allele was present in one patient.CONCLUSIONS:This is the first report of LA-related IAS in Caucasians who possess the HLA-DRB1*04:03 allele, implicating this allele in the genetic susceptibility to IAS in Caucasians. The greater occurrence of the HLA-DRB1*04:03 allele in Caucasian and other populations, combined with the growing use of LA in developed countries, may be a future predictor of additional cases of IAS.

The ubiquitous presence of insulin resistance cannot be understated. First brought to light by Himsworth and Kerr (1) in 1939, insulin resistance, defined as a subnormal response to a given dose of insulin, was ushered into prime time by Gerald Reaven (2), where it has remained at center stage. Insulin resistance is a major feature of type 2 diabetes (2). Insulin resistance is also associated with obesity, essential hypertension, dyslipidemia, nonalcoholic fatty liver disease, obstructive sleep apnea, and cancer (3). This cluster of maladies has been termed by Reaven as the “insulin resistance syndrome.” Therefore, an individual with insulin resistance is strongly predisposed to an increased risk of life-threatening clinical conditions, including cardiovascular disease. As Reaven points out, the clinical consequences of insulin resistance are not due to insulin resistance per se but come from the hyperinsulinemia that occurs as the individual with insulin resistance attempts to maintain normoglycemia. In reality, compensatory hyperinsulinemia is akin to cutting a deal with the devil. Chronic hyperinsulinemia may be beneficial to resistant tissues requiring it, for example to maintain insulin action in liver, muscle, and adipose tissues; however, it may wreak havoc with tissues that have normal sensitivity to insulin. Even within the same tissue, some of the insulin-regulated pathways, such as the glucose metabolic pathway, are more resistant to insulin than others, including the mitogenic pathway (4). Thus, it is likely that chronic overstimulation of the mitogenic pathway by insulin also plays a causative role in mediating the clinical consequences of insulin resistance. Therefore, intensive efforts are being directed toward identifying novel nutritional and pharmacological approaches that improve insulin sensitivity in target tissues. Our knowledge of …

Elevated oxidized low-density lipoprotein (OxLDL) may promote inflammation, and is associated with increased risk of atherosclerotic coronary heart disease and worsening complications of diabetes mellitus. The primary objective of this study was to evaluate the efficacy of chitin-glucan (CG), alone and in combination with a potentially anti-inflammatory olive oil (OO) extract, for reducing OxLDL in subjects with borderline to high LDL cholesterol (LDL-C) levels. This 6-week, randomized, double-blind, placebo-controlled study of a novel, insoluble fiber derived from the Aspergillus niger mycelium, CG, evaluated 130 subjects free of diabetes mellitus with fasting LDL-C 3.37–4.92 mmol/l and glucose ⩽6.94 mmol/l. Participants were randomly assigned to receive CG (4.5 g/day; n=33), CG (1.5 g/day; n=32), CG (1.5 g/day) plus OO extract (135 mg/day; n=30), or matching placebo (n=35). Administration of 4.5 g/day CG for 6 weeks significantly reduced OxLDL compared with placebo (P=0.035). At the end of study, CG was associated with lower LDL-C levels relative to placebo, although this difference was statistically significant only for the CG 1.5 g/day group (P=0.019). CG did not significantly affect high-density lipoprotein cholesterol, triglycerides, glucose, insulin or F2-isoprostane levels. Adverse events did not substantively differ between treatments and placebo. In this 6-week study, CG (4.5 g/day) reduced OxLDL, an effect that might affect the risk for atherosclerosis.

Oxidized low-density lipoprotein (OxLDL) is believed to play a role in the progression of atherosclerotic coronary heart disease (CHD) and the development of diabetes complications. This randomized, double-blind, placebo-controlled study of a novel insoluble fiber derived from the mycelium Aspergillus niger , chitin-glucan (CG) (ARTINIA™), evaluated 135 patients with fasting LDL-cholesterol 130-189.9 mg/dl and fasting glucose <=125 mg/dl. Participants were randomly assigned to receive CG (4.5 g/day; n=34), CG (1.5 g/day; n=33), CG (1.5 g/day) plus olive extract (n=33), or matching placebo (n=35) for 6 weeks. The primary outcome measure was the between-group difference in OxLDL. Secondary outcome measurements included effects upon lipid, glucose, insulin, and F2-isoprostane levels. After 6 weeks, CG 4.5 g/day (CG-4.5) significantly reduced mean OxLDL 3.8 U/L compared to baseline (58.0 U/L vs 61.8 U/L, respectively; P =0.006), and reduced OxLDL 4.97 U/L compared to placebo (P=<0.05). Other treatment groups generally had no significant effect upon OxLDL. CG treatment groups reduced LDL-cholesterol levels 3.2–;6.5% compared to placebo (P<0.05). In this study population without diabetes mellitus or elevated glucose levels, CG did not significantly affect high density lipoprotein cholesterol, triglycerides, glucose, insulin, F2-isoprostanes, or the homeostasis model assessment of insulin resistance. Treatments were well tolerated and with adverse experiences comparable to placebo. These results suggest that chitin-glucan, a novel insoluble fiber, may significantly reduce OxLDL and LDL-cholesterol levels, which may have therapeutic implications for patients at risk for CHD or other diabetes complications.

There is a worldwide pandemic of chronic cardiovascular disease. The purpose of this article is to highlight some promising nutrients present in the Mediterranean Diet that have been shown in clinical trials to reduce oxidized low-density lipoprotein (OxLDL), a pro-atherogenic mediator and biomarker for cardiovascular disease.

Bethany P. Cummings, Kimber L. Stanhope, James L. Graham, Joseph L. Evans, Denis G. Baskin, Steven C. Griffen, and Peter J. Havel Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, Davis, California; Department of Nutrition, University of California, Davis, Davis, California; ReceptorBio, Redwood City, California; Research and Development Service, Department of Veterans Affairs Puget Sound Health Care System, Seattle, Washington; and Department of Medicine, Division of Metabolism, Endocrinology, and Nutrition, University of Washington, Seattle, Washington; and Department of Internal Medicine, University of California, Davis; Sacramento, California

Sustained fructose consumption has been shown to induce insulin resistance and glucose intolerance, in part, by promoting oxidative stress. Alpha-lipoic acid (LA) is an antioxidant with insulin-sensitizing activity. The effect of sustained fructose consumption (20% of energy) on the development of T2DM and the effects of daily LA supplementation in fructose-fed University of California, Davis-Type 2 diabetes mellitus (UCD-T2DM) rats, a model of polygenic obese T2DM, was investigated. At 2 mo of age, animals were divided into three groups: control, fructose, and fructose + LA (80 mg LA.kg body wt(-1).day(-1)). One subset was followed until diabetes onset, while another subset was euthanized at 4 mo of age for tissue collection. Monthly fasted blood samples were collected, and an intravenous glucose tolerance test (IVGTT) was performed. Fructose feeding accelerated diabetes onset by 2.6 +/- 0.5 mo compared with control (P < 0.01), without affecting body weight. LA supplementation delayed diabetes onset in fructose-fed animals by 1.0 +/- 0.7 mo (P < 0.05). Fructose consumption lowered the GSH/GSSG ratio, while LA attenuated the fructose-induced decrease of oxidative capacity. Insulin sensitivity, as assessed by IVGTT, decreased in both fructose-fed and fructose + LA-supplemented rats. However, glucose excursions in fructose-fed LA-supplemented animals were normalized to those of control via increased glucose-stimulated insulin secretion. Fasting plasma triglycerides were twofold higher in fructose-fed compared with control animals at 4 mo, and triglyceride exposure during IVGTT was increased in both the fructose and fructose + LA groups compared with control. In conclusion, dietary fructose accelerates the onset of T2DM in UCD-T2DM rats, and LA ameliorates the effects of fructose by improving glucose homeostasis, possibly by preserving beta-cell function.

BACKGROUND:The purpose of this study was to determine whether a preparation of controlled-release alpha lipoic acid (CRLA) influences features of the polycystic ovary syndrome (PCOS).METHODS:We administered CRLA 600 mg twice daily for 16 weeks to six lean, nondiabetic patients with PCOS. Insulin sensitivity was measured by the euglycemic, hyperinsulinemic clamp. Plasma lipids were measured by vertical ultracentrifugation. Oxidative stress markers were measured in serum.RESULTS:At the end of 16 weeks of CRLA treatment, there was a 13.5% improvement in insulin sensitivity as determined by the euglycemic, hyperinsulinemic clamp (p < .03). There was also a lowering of triglyceride levels (p < .04) and a shift in the distribution of low-density lipoprotein (LDL) particles toward the larger, more buoyant LDL subclass fraction. Two of the subjects who were not on oral contraception had an increased number of menstrual cycles. Controlled-release alpha lipoic acid treatment, however, was neither associated with an increase in plasma antioxidant capacity nor with a reduction in plasma lipid oxidation products.CONCLUSIONS:These data suggest that the CRLA has positive effects on the PCOS phenotype. The effects of CRLA, however, may have been exerted through a mechanism not involving changes in oxidative stress.