Erika Most

Regulation of the insulin antagonistic protein tyrosine phosphatase 1B by dietary Se studied in growing rats

Protein tyrosine phosphatase 1B (PTP1B) is a key enzyme in the counterregulation of insulin signaling, and its physiological modulation depends on H2O2 and glutathione (GSH). Se via GSH peroxidases (GPxs) and its specific metabolism is involved in the removal of H2O2 and in the regulation of GSH metabolism. Recent results from animal trials and epidemiological studies with humans have shown that a high GPx1 activity or a permanent surplus of Se may promote the development of obesity and diabetes. Our nutrition physiological study with 7 x 7 growing rats was carried out to examine if PTP1B is modulated by Se supplements and, thus, may represent one trigger mediating these undesirable metabolic effects of Se. One group of rats was fed an Se-deficient diet for 8 weeks. The diets of the other six groups contained Se as selenite or selenate according to the recommendations (0.20 mg/kg diet) and at two supranutritional levels (1.00 and 2.00 mg/kg diet). All Se-supplemented animals featured a significantly higher body weight (6-14%) compared to their Se-deficient companions. Expression and activity of GPx1 in the liver of Se supplemented animals was 10- and 70-fold higher compared to Se deficiency. The detailed study of PTP1B regulation using an enzymatic assay and Western Blot analysis with an antibody against protein glutathionylation revealed that PTP1B was significantly up-regulated by both a maximization of GPx1 activity and by increasing dietary Se supply, reducing its inhibition via glutathionylation. Selenate effected a stronger PTP activation compared to selenite. In conclusion, our results suggest that the modulation of PTP1B activity may represent one plausible mechanism by which a long-term intake of Se supplements exceeding the requirements can promote the development of obesity and diabetes and needs further intensive investigation.

Exercise affects tissue lymphocyte apoptosis via redox-sensitive and Fas-dependent signaling pathways

Intensive and exhaustive exercise induces an activation of blood T-lymphocytes, which seems to be terminated by apoptotic processes in the postexercise period. Here, we report that exercise-induced T-lymphocyte apoptosis is a systemic phenomenon occurring in various lymphoid and nonlymphoid tissues. The apoptosis rate could be related to exercise intensity and type. Although in some tissues, such as the spleen and Peyers patches, an early start of apoptosis (1-3 h postexercise) could be detected, a delayed apoptosis (24 h postexercise) was observed in lung, bone marrow, and lymph nodes. Further analysis showed a similar apoptosis distribution among lymphocyte subpopulations. We tested whether components of the extrinsic or the intrinsic apoptotic pathways or both were involved in these processes. Elevated levels of lipid peroxidation-product malondialdehyde (MDA), indicating an increased production of reactive oxygen species (ROS), were found after exercise in Peyers patches, lung, and spleen, but not in lymph nodes. Application of N-acetyl-cysteine (NAC) prevented exercise-induced T-cell apoptosis completely in spleen and bone marrow, partially in lung and Peyers patches, while it was ineffective in lymph nodes. Additionally, exercise addressed the Fas-mediated apoptosis. The percentage of Fas-receptor (Fas+) and Fas-ligand positive (FasL+) lymphocytes was enhanced in Peyers patches after exercise. Moreover, FasL+ T cells were increased in the lung, while in lymph nodes Fas+ cells were increased. The critical role of Fas signaling in exercise-induced apoptosis was supported by using Fas-deficient MRL/lpr-mice. In Fas-deficient mice, exercise-induced T-lymphocyte apoptosis was prevented in spleen, lung, bone marrow, and lymph nodes, but not in Peyers patches. These data demonstrate that exercise-induced lymphocyte apoptosis is a transient systemic process with tissue-type specific apoptosis-inducing mechanisms, whose relevance for the adaptive immune competence remains to be shown.

Supplementation of a grape seed and grape marc meal extract decreases activities of the oxidative stress-responsive transcription factors NF-κB and Nrf2 in the duodenal mucosa of pigs

BackgroundIn pigs, enteric infections and the development of gut disorders such as diarrhoea are commonly observed, particularly after weaning. The present study investigated the hypothesis that feeding a grape seed and grape marc extract (GSGME) as a dietary supplement has the potential to suppress the inflammatory process in the small intestine of pigs by modulating the activities of NF-κB and Nrf2 due to its high content of flavonoids.MethodsTwenty-four crossbred, 6 weeks old pigs were randomly assigned to 2 groups of 12 animals each and fed nutritionally adequate diets without or with 1% GSGME for 4 weeks.ResultsPigs administered GSGME had a lower transactivation of NF-κB and Nrf2 and a lower expression of various target genes of these transcription factors in the duodenal mucosa than control pigs (P < 0.05). Concentrations of α-tocopherol and thiobarbituric acid reactive substances (TBARS) in liver and plasma and total antioxidant capacity of plasma and relative mRNA abundances of NF-κB and Nrf2 target genes in the liver did not differ between the two groups. However, the ratio of villus height:crypt depth and the gain:feed ratio was higher in the pigs fed GSGME than in control pigs (P < 0.05).ConclusionsThis study shows that dietary supplementation of a polyphenol rich GSGME suppresses the activity of NF-κB in the duodenal mucosa of pigs and thus might provide a useful dietary strategy to inhibit inflammation in the gut frequently occurring in pigs. Feeding GSGME did not influence vitamin E status and the antioxidant system of the pigs but improved the gain:feed ratio. In overall, the study suggests that polyphenol-rich plant extracts such GSGME could be useful feed supplements in pig nutrition, in order to maintain animal health and improve performance.

Dietary effect of phytogenic phytase and an addition of microbial phytase to a diet based on field beans, wheat, peas and barley on the utilization of phosphorus, calcium, magnesium, zinc and protein in piglets

SummaryThe effect of the addition of microbial phytase to a diet based on field beans (30 %), wheat (28 %), peas (25 %), and barley (14 %) was studied in a 2-week experiment with 3×8 castrated male, individually housed, hybrid piglets (live weight range 12–16 kg). All diets contained about 4.7 g Ca, 4.2 g P (77 % present as phytate phosphorus), 1.0 g Mg, 60 mg Zn per kg diet, and 17 % crude protein. Group I was fed the basal diet with a native phytase-activity of about 260 U per kg diet. In group II, 350 U, in group III, 700 U of microbial phytase per kg diet were added. The addition of microbial phytase improved the apparent P absorption (% of intake) from 48 % (group I) to 66 % (group II) and 71 % (group III). Comparable positive effects from the phytase treatment were obtained for the calcium utilization. The phytase supplementation also enhanced plasma zinc concentration significantly. The concentration of inorganic phosphorus in plasma, the zinc digestibility, and the magnesium balance were improved in tendency. The utilization of nitrogen remained unchanged.ZusammenfassungIn einem zweiwöchigen Stoffwechselversuch mit 3×8 männlichen kastrierten Hybridferkeln (Lebendmassebereich 12–16 kg) wurde die Wirkung einer Zulage an mikrobieller Phytase zu einer Diät auf der Basis von Ackerbohnen (30 %), Weizen (28 %), Futtererbsen (25 %) und Gerste (14 %) untersucht. Die Diäten enthielten je kg Diät ca. 4,7 g Ca, 4,2 g P (davon ca. 77 % Phytin-P), 1,0 g Mg, 60 mg Zn sowie 17 % Rohprotein. Zum nativen Phytasegehalt von ca. 260 U/kg Diät (Gruppe I) wurden in den Gruppen II und III 350 bzw. 700 U mikrobieller Phytase/kg Diät supplementiert. Durch die Zulage von 350 und 700 U mikrobieller Phytase je kg Diät wurde die scheinbare P-Absorption (in % der Aufnahme) von 48 % in der Kontrollgruppe (Gruppe I) auf 66 % (Gruppe II) und 71 % (Gruppe III) gesteigert. Ähnlich positive Effekte traten bei der Ca-Verwertung auf. Auch die Plasma-Zn-Konzentration wurde durch die Phytasesupplementierung signifikant gesteigert. Die Gehalte an anorganischem Phosphat des Blutplasmas, die scheinbare Zn-Absorption und die Magnesiumverwertung wurden durch die Phytasezulage in der Tendenz erhöht, während die N-Bilanz unbeeinflußt blieb.

APOE ε4 is associated with higher vitamin D levels in targeted replacement mice and humans

The allele ε4 of apolipoprotein E (APOE), which is a key regulator of lipid metabolism, represents a risk factor for cardiovascular diseases and Alzheimers disease. Despite its adverse effects, the allele is common and shows a nonrandom global distribution that is thought to be the result of evolutionary adaptation. One hypothesis proposes that the APOE ε4 allele protects against vitamin D deficiency. Here we present, for the first time, experimental and epidemiological evidence that the APOE ε4 allele is indeed associated with higher serum vitamin D [25(OH)D] levels. In APOE4 targeted replacement mice, significantly higher 25(OH)D levels were found compared with those in APOE2 and APOE3 mice (70.9 vs. 41.8 and 27.8 nM, P<0.05). Furthermore, multivariate adjusted models show a positive association of the APOE ε4 allele with 25(OH)D levels in a small collective of human subjects (n=93; P=0.072) and a general population sample (n=699; P=0.003). The novel link suggests ε4 as a modulator of vitamin D status. Although this result agrees well with evolutionary aspects, it appears contradictory with regard to chronic diseases, especially cardiovascular disease. Large prospective cohort studies are now needed to investigate the potential implications of this finding for chronic disease risks.—Huebbe, P., Nebel, A., Siegert, S., Moehring, J., Boesch‐Saadatmandi, C., Most, E., Pallauf, J., Egert, S., Müller, M. J., Schreiber, S., Nöthlings, U., Rimbach, G. APOE ε4 is associated with higher vitamin D levels in targeted replacement mice and humans. FASEB J. 25, 3262‐3270 (2011). www.fasebj.org

Effects of dietary polyphenol-rich plant products from grape or hop on pro-inflammatory gene expression in the intestine, nutrient digestibility and faecal microbiota of weaned pigs

BackgroundFeeding polyphenol-rich plant products has been shown to increase the gain:feed ratio in growing pigs. The reason for this finding has not yet been elucidated. In order to find the reasons for an increase of the gain:feed ratio, this study investigated the effect of two polyphenol-rich dietary supplements, grape seed and grape marc meal extract (GSGME) or spent hops (SH), on gut morphology, apparent digestibility of nutrients, microbial composition in faeces and the expression of pro-inflammatory genes in the intestine of pigs.ResultsPigs fed GSGME or SH showed an improved gain:feed ratio in comparison to the control group (P < 0.10 for GSGME, P < 0.05 for SH). Villus height:crypt depth ratio in duodenum and jejunum as well as apparent total tract digestibility of nutrients were unchanged in the groups receiving GSGME or SH in comparison to the control group. However, the groups receiving GSGME or SH revealed an increased faecal pH value, lower levels of volatile fatty acids and lower counts of Streptococcus spp. and Clostridium Cluster XIVa in the faecal microbiota (P < 0.05). Moreover, both treatment groups had a lower expression of various pro-inflammatory genes in duodenum, ileum and colon than the control group (P < 0.05).ConclusionThe present study suggests that dietary plant products rich in polyphenols are able to improve the gain:feed ratio in growing pigs. It is assumed that an alteration in the microbial composition and anti-inflammatory effects of the polyphenol-rich plant products in the intestine might contribute to this effect.

Effect of Phytic Acid and Microbial Phytase on Cd Accumulation, Zn Status, and Apparent Absorption of Ca, P, Mg, Fe, Zn, Cu, and Mn in Growing Rats

Three groups of individually housed albino rats (n = 6, initial average weight = 47 g) were fed diets based on egg white and cornstarch (basal diet 8 g Ca, 5.2 g P, 0.76 g Mg, 100 mg Zn, 100 mg Fe, 50 mg Mn, 7 mg Cu, and 5 mg Cd per kilogram diet) over a 4-week period. Group I (controls) was fed the basal diet free of phytic acid (PA) and microbial phytase. In groups II and III cornstarch was replaced by 0.5% PA from NaPA (molar PA/Zn ratio approximately 5). In group III, 2,000 U of microbial phytase from Aspergillus niger per kilogram diet was added. Live weight gain, zinc status (zinc in plasma, femur, liver, and testes; activity of the plasma alkaline phosphatase), and apparent absorption of zinc, iron, copper, and manganese remained unchanged by the different dietary treatments. The apparent phosphorus absorption was highest in the phytase group. PA decreased and microbial phytase improved the apparent absorption of calcium and magnesium. Liver cadmium concentration, total liver and kidney cadmium content, as well as fractional liver and kidney cadmium accumulation in rats fed the diet containing PA were significantly higher than those in the controls. Phytase supplementation lowered liver and kidney cadmium accumulation. Differences in calcium and magnesium bioavailability due to PA and microbial phytase may be one factor in the alteration of tissue cadmium accumulation.

Supplemental Phytic Acid and Microbial Phytase Change Zinc Bioavailability and Cadmium Accumulation in Growing Rats

Three groups of individually housed albino rats (n = 6 each, initial average weight = 47 g) were fed diets based on egg white and corn starch over a 4-week period. All diets were supplemented with 15 mg/kg of Zn and 5 mg/kg of Cd. Group I (Control) was fed the basal diet free of phytic acid (PA) and phytase. By replacing corn starch by 0.5% PA (as NaPA) in groups II and III, a molar PA/Zn ratio of 33 was obtained. In group III, 2000 U of microbial phytase per kg diet were added. Addition of PA to diet (group II) resulted in a significant decrease in growth and zinc status. The negative effect of dietary PA on growth and zinc status was considerably counteracted by the supplementation of 2000 U microbial phytase (group III). In group I the highest apparent zinc absorption (58.2%) was measured. The addition of 0.5% PA (group II) significantly decreased apparent zinc absorption to 23.4%. In rats receiving the phytase-enriched diet (group III) 46.5% of ingested zinc was apparently absorbed. Liver cadmium concentration in rats fed the diet containing PA was significantly higher than that in the control group, whereas phytase supplementation lowered liver cadmium accumulation. In tendency similar effects were obtained for kidney cadmium accumulation.

Parameters of dietary selenium and vitamin E deficiency in growing rabbits

4 x 5 growing female rabbits (New Zealand White) with an initial live weight of 610 +/- 62 g were fed a torula yeast based semisynthetic diet low in selenium (<0.03 mg/kg diet) and containing <2 mg alpha-tocopherol per kg (group I). Group II received a vitamin E supplementation of 150 mg alpha-tocopherylacetate per kg diet, whereas for group III 0.40 mg Se as Na-selenite and for group IV both supplements were added. Selenium status and parameters of tissue damage were analyzed after 10 weeks on experiment (live weight 2,355 +/- 145 g). Selenium depletion of the Se deficient rabbits (groups I and II) was indicated by a significantly lower plasma Se content (group I: 38.3 +/- 6.23 microg Se/mL plasma, group II: 42.6 +/- 9.77, group III: 149 +/- 33.4, group IV: 126 +/- 6.45) and a significantly lower liver Se content (group I: 89.4 +/- 18.2 microg/kg fresh matter, group II: 111 +/- 26.2) as compared to the Se supplemented groups III (983 +/- 204) and IV (926 +/- 73.9). After 5 weeks on the experimental diets differences in the development of plasma glutathione peroxidase were observed. As compared to the initial status group (45.2 +/- 4.50) pGPx activity in mU/mg protein was decreased in group I (19.1 +/- 7.08), remained almost stable in the vitamin E supplemented group II (46.3 +/- 11.2) whereas an elevated enzyme activity was measured in the Se supplemented groups III (62.4 +/- 23.9) and IV (106 +/- 19.9). In the rabbit organs investigated 10 weeks of Se deficiency caused a significant loss of Se dependent cellular glutathione peroxidase activity (GPx1) of 94% (liver), 80% (kidney), 50% (heart muscle) and 60% (musculus longissimus dorsi) in comparison to Se supplemented control animals. Damage of cellular lipids and proteins in the liver was due to either Se or vitamin E deficiency. However damage was most severe under conditions of a combined Se and vitamin E deficiency. It can be concluded that the activity of plasma glutathione peroxidase is a sensitive indicator of Se deficiency in rabbits. The loss of GPx1 activity indicates the selenium depletion in various rabbit organs. Both selenium and vitamin E are essential and highly efficient antioxidants which protect rabbits against lipid and protein oxidation.

Supplementation of carnitine leads to an activation of the IGF-1/PI3K/Akt signalling pathway and down regulates the E3 ligase MuRF1 in skeletal muscle of rats

BackgroundRecently, it has been shown that carnitine down-regulates genes involved in the ubiquitin-proteasome system (UPS) in muscle of pigs and rats. The mechanisms underlying this observation are yet unknown. Based on the previous finding that carnitine increases plasma IGF-1 concentration, we investigated the hypothesis that carnitine down-regulates genes of the UPS by modulation of the of the IGF-1/PI3K/Akt signalling pathway which is an important regulator of UPS activity in muscle.MethodsMale Sprague–Dawley rats, aged four weeks, were fed either a control diet with a low native carnitine concentration or the same diet supplemented with carnitine (1250 mg/kg diet) for four weeks. Components of the UPS and IGF-1/PI3K/Akt signalling pathway in skeletal muscle were examined.ResultsRats fed the diet supplemented with carnitine had lower mRNA and protein levels of MuRF1, the most important E3 ubiquitin ligase in muscle, decreased concentrations of ubiquitin-protein conjugates in skeletal muscle and higher IGF-1 concentration in plasma than control rats (P < 0.05). Moreover, in skeletal muscle of rats fed the diet supplemented with carnitine there was an activation of the PI3K/Akt signalling pathway, as indicated by increased protein levels of phosphorylated (activated) Akt1 (P < 0.05).ConclusionThe present study shows that supplementation of carnitine markedly decreases the expression of MuRF1 and concentrations of ubiquitinated proteins in skeletal muscle of rats, indicating a diminished degradation of myofibrillar proteins by the UPS. The study moreover shows that supplementation of carnitine leads to an activation of the IGF-1/PI3K/Akt signalling pathway which in turn might contribute to the observed down-regulation of MuRF1 and muscle protein ubiquitination.