Vitamin E: Curse or Benefit in Alzheimer's Disease? A Systematic Investigation of the Impact of α-, γ- and δ-Tocopherol on Aß Generation and Degradation in Neuroblastoma Cells.

OBJECTIVES: The E vitamins are a class of lipophilic compounds including tocopherols, which have high antioxidative properties. Because of the elevated lipid peroxidation and increased reactive oxidative species in Alzheimer's disease (AD) many attempts have been made to slow down the progression of AD by utilizing the antioxidative action of vitamin E. Beside the mixed results of these studies nothing is known about the impact of vitamin E on the mechanisms leading to amyloid-ß production and degradation being responsible for the plaque formation, one of the characteristic pathological hallmarks in AD. Here we systematically investigate the influence of different tocopherols on Aß production and degradation in neuronal cell lines. MEASUREMENTS: Beside amyloid-ß level the mechanisms leading to Aß production and degradation are examined. RESULTS: Surprisingly, all tocopherols have shown to increase Aß level by enhancing the Aß production and decreasing the Aß degradation. Aß production is enhanced by an elevated activity of the involved enzymes, the ß- and γ-secretase. These secretases are not directly affected, but tocopherols increase their protein level and expression. We could identify significant differences between the single tocopherols; whereas α-tocopherol had only minor effects on Aß production, δ-tocopherol showed the highest potency to increase Aß generation. Beside Aß production, Aß clearance was decreased by affecting IDE, one of the major Aß degrading enzymes. CONCLUSIONS: Our results suggest that beside the beneficial antioxidative effects of vitamin E, tocopherol has in respect to AD also a potency to increase the amyloid-ß level, which differ for the analysed tocopherols. We therefore recommend that further studies are needed to clarify the potential role of these various vitamin E species in respect to AD and to identify the form which comprises an antioxidative property without having an amyloidogenic potential.

Effects of neuron-specific ADAM10 modulation in an in vivo model of acute excitotoxic stress.

A disintegrin and metalloprotease (ADAM) 10 is the main candidate enzyme for the alpha-secretase processing of the amyloid precursor protein (APP). Neuron-specific ADAM10 overexpression proved beneficial in the APP[V717I] mutant Alzheimer mouse model [Postina R, Schroeder A, Dewachter I, Bohl J, Schmitt U, Kojro E, Prinzen C, Endres K, Hiemke C, Blessing M, Flamez P, Dequenne A, Godaux E, van Leuven F, Fahrenholz F (2004) A disintegrin-metalloproteinase prevents amyloid plaque formation and hippocampal defects in an Alzheimer disease mouse model. J Clin Invest 113:1456-1464]. Since Alzheimer patients have a high prevalence for epileptic seizures, we investigated the effects of ADAM10 modulation under conditions of experimentally induced epileptic seizures. In this context we also examined whether ADAM10 effects were influenced by APP levels. Therefore we compared severity of kainate-induced seizures, neurodegeneration and inflammation in double transgenic mice overexpressing functional ADAM10 or a dominant negative ADAM10 mutant in the APP[V717I] background with single transgenic ADAM10 modulated mice. Double transgenic dominant negative ADAM10dn/APP[V717I] mice suffered from stronger epileptic seizures, had a longer recovery period and showed more neurodegeneration and glial activation in the hippocampal region than double transgenic mice moderately overexpressing functional ADAM10 (ADAM10mo/APP[V717I]) and APP[V717I] mice with endogenous ADAM10 levels. This suggests that ADAM10 activity is necessary to provide neuroprotection against excitotoxicity in the APP[V717I] mouse model. Interestingly, increased expression of functional ADAM10 above the endogenous level did not correlate with a better protection against seizures and neurodegeneration. Furthermore, ADAM10 dominant negative mice without transgenic APP overexpression (ADAM10dn) were seizing for a shorter time and showed less neuronal cell death and neuroinflammation after kainate injection than wild-type mice, which shows beneficial effects of ADAM10 inhibition in context with neurodegeneration. In contrast, mice with a high ADAM10 overexpression showed more seizures and stronger neuronal damage and inflammation than wild-type mice and mice with moderate ADAM10 overexpression. Hence, additional cleavage products of ADAM10 may counterbalance the neuroprotective effect of alpha-secretase-cleaved APP in the defense against excitotoxicity. Our findings highlight the need of a careful modulation of ADAM10 activity for neuroprotection depending on substrate availability and on neurotoxic stress conditions.

Effects of 9,10 anthraquinone on ruminal fermentation, total-tract digestion, and blood metabolite concentrations in sheep.

The objective of this study was to evaluate the effects of adding 9,10 anthraquinone, a known inhibitor of methanogenesis and sulfate reduction, on blood metabolites, digestibility, and distribution of gas in sheep. In all experiments, we fed a complete pelleted diet that contained 17.5% crude protein and 24.5% acid detergent fiber. In an 8-wk study, feeding up to 66 ppm (dry matter basis) of 9,10 anthraquinone had no adverse effects on blood metabolites including indicators of normal enzyme function, mineral concentrations, and hematological measurements. Feeding 9,10 anthraquinone had no effect on average daily gain, although sheep fed a diet containing 66 ppm of 9,10 anthraquinone numerically gained the least weight. The ruminal molar proportions of acetic acid were decreased (P < 0.05) and the molar proportions of propionic acid were increased (P < 0.05) in sheep fed 1.5 and 66 ppm 9,10 anthraquinone when compared to those fed an unsupplemented diet. In a digestion trial, 9,10 anthraquinone (33 and 66 ppm) had no effect on the apparent digestion of nutrients in the total gastrointestinal tract. In a metabolism study, ruminal gasses were collected by rumenocentesis and analyzed for methane and hydrogen concentrations. Feeding 500 ppm of 9,10 anthraquinone to sheep resulted in a decrease (P < 0.07) in the concentration of methane, but an increase (P < 0.05) in hydrogen concentration of ruminal gas throughout the 19 d of feeding. There was no indication of ruminal adaptation throughout this time. These results are the first to show that 9,10 anthraquinone can partially inhibit in vivo rumen methanogenesis, which supports previous in vitro findings. In addition, at the concentrations used in this study, 9,10 anthraquinone was not toxic to ruminants.

The effect of treating forages with fibrolytic enzymes on its nutritive value and lactation performance of dairy cows.

Forages (corn silage and alfalfa hay) were sprayed with liquid enzymes prior to combining with a concentrate to form a total mixed ration (50% forage:50% concentrate, dry matter basis) and fed to lactating cows. In the first year, treatments were 1) no enzymes, 2) an enzyme complex containing 3500 carboxymethyl cellulase (CMCase) and 16,000 xylanase units per kilogram of forage dry matter, or 3) an enzyme complex containing 8800 CMCase units and 40,000 xylanase units. In the second year, the treatments were 1) no enzymes, 2) an enzyme complex as in yr 1 containing 3700 CMCase and 14,000 xylanase units, or 3) an enzyme complex using an alternative cellulase and containing 3600 CMCase and 11,000 xylanase units. In the first year, cows fed diet 2 tended to produce more milk (39.5 kg/d) than those fed diet 1 (37.0 kg/d) or those fed diet 3 (36.2 kg/d). The high level of enzyme treatment in diet 3 decreased the output of milk protein and fat compared to the low level of enzyme treatment. In the second year, cows fed diet 3 produced more milk (35.4 kg/d) than did those fed diet 1 (32.9 kg/d) and numerically more than those fed diet 2 (33.6 kg/d). Milk fat and protein were similar among treatments but numerically lower for cows fed enzyme-treated forages. Dry matter intake (kg/d) was similar among treatments in both years. Spraying certain doses and combinations of enzymes directly onto forages prior to feeding can improve milk yields but enzyme sources and dose levels are of critical importance.

The effect of preservatives based on propionic acid on the fermentation and aerobic stability of corn silage and a total mixed ration.

For 3 successive yr, whole-plant corn was ensiled in laboratory silos with low percentages of silage preservatives, the primary active ingredient of which was propionic acid. Preservatives were added to forage just prior to ensiling at rates of 0.1 to 0.2% of the fresh forage weight. In all 3 yr, treatments had minor effects on fermentation end products, except that the concentration of propionic acid was greater because of its addition. The mean low and high percentages of preservatives increased aerobic stability of the treated silages by 19 and 57 h, respectively, in Experiment 1 and by 17 and 38 h, respectively, in Experiment 2. In Experiment 3, aerobic stability was improved by > 90 h by preservatives (0.2% addition). In a lactation study, a total mixed ration (46% dry matter) was mixed without or with (0.2 or 0.3%) a stabilizer that was designed to prevent spoilage in the feed bunk. The high dose resulted in orts with a lower pH and temperature after 24 h in the feed bunk. However, dry matter intake and milk production were unaffected by treatments. Chemical preservatives based on propionic acid added at low rates did not affect fermentation but were effective in the reduction of heating in corn silage and in a total mixed ration.