Cellular signaling in pseudoxanthoma elasticum: an update.

Pseudoxanthoma elasticum is an autosomal recessive genodermatosis with variable expression, due to mutations in the ABCC6 or ENPP1 gene. It is characterized by elastic fiber mineralization and fragmentation, resulting in skin, eye and cardiovascular symptoms. Significant advances have been made in the last 20 years with respect to the phenotypic characterization and pathophysiological mechanisms leading to elastic fiber mineralization. Nonetheless, the substrates of the ABCC6 transporter - the main cause of PXE - remain currently unknown. Though the precise mechanisms linking the ABCC6 transporter to mineralization of the extracellular matrix are unclear, several studies have looked into the cellular consequences of ABCC6 deficiency in PXE patients and/or animal models. In this paper, we compile the evidence on cellular signaling in PXE, which seems to revolve mainly around TGF-ßs, BMPs and inorganic pyrophosphate signaling cascades. Where conflicting results or fragmented data are present, we address these with novel signaling data. This way, we aim to better understand the up- and down-stream signaling of TGF-ßs and BMPs in PXE and we demonstrate that ANKH deficiency can be an additional mechanism contributing to decreased serum PPi levels in PXE patients.

Growth response, blood characteristics and copper accumulation in organs of broilers fed on diets supplemented with organic and inorganic dietary copper sources.

1. A 56-d experiment was conducted to study the comparative influence of organic and inorganic dietary copper (Cu) sources on growth, blood characteristics and copper accumulation in organs of broilers. 2. A total of 480 Arbor-Acre unsexed broilers were fed on diets containing copper sulphate (CuSO(4)) or copper proteinate (Cu Pro) at concentrations of 50, 100 or 150 mg/kg of Cu supplementation. The birds were given a broiler starter diet from 1-28 d and a broiler finisher diet from 29-56 d which contained 30·8 mg/kg and 41·1 mg/kg basal copper concentration respectively. Growth performance, blood characteristics and Cu accumulation in organs of the broilers were measured. 3. At 28 d, Cu Pro-fed birds had improved feed conversion ratio compared with CuSO(4). At 56 d, birds fed on Cu Pro diets had significantly greater body weight than CuSO(4)-fed birds. Birds fed on CuSO(4) supplemented diets had significantly better feed conversion efficiency. Feed consumptions for the two Cu sources were not significantly different. At no stage did the concentration of added Cu affect the productive traits measured. 4. Cu Pro supplementation increased haemoglobin concentration but reduced plasma triglyceride and plasma cholesterol. Plasma cholesterol decreased as Cu concentration increased. 5. There was a greater accumulation of Cu in the blood, heart, lung, liver and bone of broilers fed on Cu Pro than in those receiving CuSO(4). The liver Cu concentration increased as dietary Cu concentration increased. 6. Cu Pro was more effective in promoting growth and reducing blood cholesterol, and was more bio-available in the organs of broilers.

Influence of a synbiotic mixture consisting of Lactobacillus acidophilus 74-2 and a fructooligosaccharide preparation on the microbial ecology sustained in a simulation of the human intestinal microbial ecosystem (SHIME reactor).

Lactobacillus acidophilus 74-2, which is used in probiotic products, was administered, with fructo-oligosaccharide in a milk-based product, to the second vessel (duodenum/jejunum) of the SHIME reactor, an in vitro simulation of the human intestinal microbial ecology. The main focus of this study was to monitor the changes of the population density of selected bacterial species in the intestine and the changes of metabolic activities during the supplementation of L. acidophilus and fructooligosaccharide in the SHIME reactor. Interestingly, the addition of L. acidophilus 74-2 with fructooligosaccharide gave rise to an increase of bifidobacteria. Moreover, major positive changes occurred in the production of volatile fatty acids: a strong upward trend was observed especially in the case of butyric acid and propionic acid. Furthermore a noticeable increase of beta-galactosidase activity was monitored, while the activity of beta-glucuronidase, generally considered undesirable, declined.

The effect of probiotic strains on the microbiota of the Simulator of the Human Intestinal Microbial Ecosystem (SHIME).

The aim of the present work was to study five potential probiotic strains (Lactobacillus plantarum, two strains of L. paracasei subsp. paracasei, L. rhamnosus and Bifidobacterium sp.) comparatively in the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) in vitro model, and to evaluate this model as a tool in the screening and selection of probiotic bacteria. The impact of the strains on the composition of microbiota and its metabolic activities (production of lactic acid and short-chain fatty acids) was studied. Changes in composition of the microbiota become apparent as a result of probiotic treatment. A marked, but temporary, increase was noted in the number of lactic acid bacteria and bifidobacteria. The profiles of D(-) and L(+) isomers of lactic acid detected in the SHIME after addition of probiotic strains corresponded well to those that are produced in pure culture conditions. The numbers of enterobacteriaceae decreased markedly and those of clostridia detectably during the intervention, while the enterococci tended to increase after the treatment. This pattern was similar in the reactors representing both the small and large intestine in the model. The changes in short-chain fatty acids were small, and no definite trend was observed.

The colonization of a simulator of the human intestinal microbial ecosystem by a probiotic strain fed on a fermented oat bran product: effects on the gastrointestinal microbiota.

The effects of Lactobacillus-GG-fermented oat bran product on the microbiota and its metabolic activity in the human gut were investigated, using a simulator of the human intestinal microbial ecosystem (SHIME), by analysing the bacterial population, shortchain fatty acids and gas production. In addition, the effects of fermented oat bran supernatant and supernatant samples from reactors 4, 5 and 6 (large intestine) on the growth of Escherichia coli IHE 13047, Enterococcus faecalis VTT E-93203, Lactobacillus rhamnosus VTT E-94522 (Lactobacillus GG) and Lactococcus lactis subsp. lactis VTT E-90414 were monitored to ascertain possible stimulatory/inhibitory effects by an in vitro turbidometric method. Our experiments showed that Lactobacillus GG colonized the SHIME reactor and this colonization could be maintained for several weeks without extra supplementation. Oat bran feeding also favoured the growth of bifidobacteria and caused an increase in the production of acetic, propionic and butyric acid as well as CH4 and CO2. However, the effects of oat bran, either on bacterial populations or on their metabolic activity, were not directly dose-dependent. In turbidometric measurements, the supernatant of fermented oat bran exerted an inhibitory effect of Lactobacillus GG, but stimulated the growth of enterococci.

Effect of the addition of Peptostreptococcus productus ATCC35244 on the gastro-intestinal microbiota and its activity, as simulated in an in vitro simulator of the human gastro-intestinal tract.

Peptostreptococcus productus ATCC35244, a reductive acetogenic strain, was added daily over 9 successive days to the fourth vessel (ascending colon) of the SHIME, a six-stage reactor system simulating the in vivo continuous culture conditions of the human gastro-intestinal tract. Final numbers of organisms (cfu)/ml reactor contents (c) were attained such that log10c = 6.9 +/- 0.1. The addition caused the CH4 production to decrease below the detection limit while total gas and CO2 production in the fifth (transverse colon) and sixth reactor (descending colon) were lowered and the acetic acid concentration was augmented. Ending the supplementation caused CH4 production to re-establish within 4 days, while CO2 production increased much more slowly. The concentration of acetic acid only started to decrease after 7 days. The results indicate that P. productus, upon regular administration, is able to compete with methanogens for H2 in the gastro-intestinal microbial ecosystem because of its reductive acetogenic character.

Effect of 2-bromoethanesulfonic acid and Peptostreptococcus productus ATCC 35244 addition on stimulation of reductive acetogenesis in the ruminal ecosystem by selective inhibition of methanogenesis.

Evidence is provided that reductive acetogenesis can be stimulated in ruminal samples during short-term (24-h) incubations when methanogenesis is inhibited selectively. While addition of the reductive acetogen Peptostreptococcus productus ATCC 35244 alone had no significant influence on CH4 and volatile fatty acid (VFA) production in ruminal samples, the addition of this strain together with 2-bromoethanesulfonic acid (BES) (final concentration, 0.01 or 0.03 mM) resulted in stimulation of acetic acid production and H2 consumption. Since acetate production exceeded amounts that could be attributed to reductive acetogenesis, as measured by H2 consumption, it was found that P. productus also fermented C6 units (glucose and fructose) heterotrophically to mainly acetate (> 99% of the total VFA). Using 14CH3COOH, we concluded that addition of BES and BES plus P. productus did not alter the consumption of acetate in ruminal samples. The addition of P. productus to BES-treated ruminal samples caused supplemental inhibition of CH4 production and stimulation of VFA production, representing a possible energy gain of about 13 to 15%.

Gastro-enteric methane versus sulphate and volatile fatty acid production.

The breakdown of low digestible components present in food during passage through the human and animal gastro-intestinal (GI) tract is performed by the highly diverse microbial community present in this ecosystem. Fermentation of these substances yields, besides CO2 and volatile fatty acids, H2, which is used as a substrate by three different H2-consuming bacteria. Sulphate-reducing bacteria (SRB) use H2 to reduce SO inf4 (sup2-) to H2S, hydrogenotrophic methane-producing bacteria (MPB) use H2 to reduce CO2 to CH4 and reductive acetogens (RAC) use H2 to reduce CO2 to CH3COOH. A competition between these three bacterial groups exists for the common H2 substrate. This results generally in the dominance of one group above the other two.