Serum levels of the adipokine chemerin are increased in preeclampsia during and 6 months after pregnancy

Preeclampsia is a serious cardiovascular complication in pregnancy which is associated with an increased future metabolic and cardiovascular risk for mother and newborn. Recently, chemerin was introduced as a novel adipokine inducing insulin resistance in vitro and in vivo. In the current study, we investigated serum concentrations of chemerin by ELISA in control and preeclampsia patients during pregnancy (Control: n=37, preeclampsia: n=37) and 6 months after delivery (Control: n=35, preeclampsia: n=36). Furthermore, the association between chemerin and markers of renal function, glucose and lipid metabolism, as well as inflammation was studied in pregnant patients. Median maternal chemerin concentrations were significantly elevated in preeclampsia patients (249.5 [range: 123.1-366.9] μg/l) as compared to controls (204.8 [138.5-280.8] μg/l) (p<0.001). Furthermore, chemerin serum levels positively correlated with blood pressure, creatinine, free fatty acids, cholesterol, triglycerides (TG), leptin, adiponectin, and C-reactive protein in univariate analyses. In multivariate analyses, TG and leptin remained independently associated with circulating chemerin. Interestingly, median chemerin concentrations 6 months after delivery remained significantly higher in former preeclampsia patients (196.0 [119.8-368.7] μg/l) as compared to controls (152.2 [102.8-216.4] μg/l). Taken together, maternal chemerin serum concentrations are significantly increased in preeclampsia during and after pregnancy. Furthermore, TG and leptin are independent predictors of circulating chemerin during pregnancy.     

A scoring system for coat and tail condition in ringtailed lemurs,Lemur catta

Coat condition can be influenced by a wide variety of disorders and thus provides a useful tool for noninvasive health and welfare assessments in wild and captive animals. Using Lemur catta as an exemplar, we offer a 6‐step scoring system for coat and tail condition, ranging from perfectly fluffy to half or more of body and tail being hairless. The categories are described in detail and illustrated with sample pictures from a wild population in Berenty Reserve, Madagascar. Furthermore, we elaborate on intermediate conditions and discoloration of fur. Coat condition scoring allows the comparison between years, seasons, and the effect of toxin, disease or stress. Although this system was developed for wild L. catta, we believe it can also be of value for other species. We recommend scoring coat condition in healthy wild mammal populations to give a baseline on yearly and seasonal variations vs. deteriorating health conditions or pathology. Am. J. Primatol. 71:183–190, 2009. © 2009 Wiley‐Liss, Inc.     

Enzymatic degradation and electrospray tandem mass spectrometry as tools for determining the structure of cationic starches prepared by wet and dry methods

Cationic starches from various semi-technical processes, two 'wet' (slurry and paste modification) and two 'dry' procedures (dry modification and extrusion), each type in a DS range from 0.03 to 0.1, were investigated by electrospray ionisation mass spectrometry (ESIMS) and tandem mass spectrometry (ESIMS2) after enzymatic degradation with alpha-amylase and subsequent glucoamylase digestion. For comparison, chemically derived cationic oligosaccharides were also analysed by ESIMS. The cationisation pattern in the glucosyl units was analysed by GLC after methanolysis, permethylation and Hofmann elimination. Results from ESIMS are discussed and interpreted with respect to enzyme susceptibility, monomer composition and physical properties of the different types of cationic starches.     

Agricultural fertilization with poultry manure results in persistent environmental contamination with the pathogen Clostridioides difficile

During a field experiment applying broiler manure for fertilization of agricultural land, we detected viable Clostridioides (also known as Clostridium) difficile in broiler faeces, manure, dust and fertilized soil. A large diversity of toxigenic C. difficile isolates was recovered, including PCR ribotypes common from human disease. Genomic relatedness of C. difficile isolates from dust and from soil, recovered more than 2 years after fertilization, traced their origins to the specific chicken farm that had delivered the manure. We present evidence of long-term contamination of agricultural soil with manure-derived C. difficile and demonstrate the potential for airborne dispersal of C. difficile through dust emissions during manure application. Clostridioides genome sequences virtually identical to those from manure had been recovered from chicken meat and from human infections in previous studies, suggesting broiler-associated C. difficile are capable of zoonotic transmission.     

Vaccenic acid-mediated reduction in cytokine production is independent of c9,t11-CLA in human peripheral blood mononuclear cells

The ruminant trans fatty acid vaccenic acid (tVA) favorably alters markers of inflammation. However, it is not yet clear whether these effects are attributed to its endogenous partial conversion to c9,t11-CLA, which is known to possess anti-inflammatory properties. We compared the cytokine reducing potential of tVA to c9,t11-CLA in human T-helper (Th) cells as a main source of cytokine production during inflammation. Secondly, we assessed whether a bioconversion of tVA to c9,t11-CLA via stearoyl-CoA desaturase (SCD) encoded activity takes place in peripheral blood mononuclear cells (PBMC) in order to relate the outcomes of intracellular cytokine measurement to the degree of conversion. TVA reduced the percentage of both IL-2 and TNF-α expressing Th cells significantly, but to a lesser extent compared to c9,t11-CLA, as determined by flow cytometry after alloreactive stimulation of PBMC. Pre-treatment with the selective PPARγ antagonist T0070907 largely re-established the IL-2 and TNF-α positive Th cell population in both tVA and c9,t11-CLA treated cultures. Interestingly, while the portion of tVA dose-dependently increased within the cellular lipid fraction, the initially marginal amount of c9,t11-CLA remained unaltered. However, SCD mRNA although abundantly expressed in PBMC was not regulated by tVA. Conclusively, these results suggest that the cytokine reducing effect of tVA in human T cells is independent of c9,t11-CLA, since no bioconversion occurred. Moreover, the data provide evidence that tVA mechanistically acts in a manner similar to c9,t11-CLA.     

Enhancement of Carotenoid Biosynthesis in Transplastomic Tomatoes by Induced Lycopene-to-Provitamin A Conversion

Carotenoids are essential pigments of the photosynthetic apparatus and an indispensable component of the human diet. In addition to being potent antioxidants, they also provide the vitamin A precursor β-carotene. In tomato (Solanum lycopersicum) fruits, carotenoids accumulate in specialized plastids, the chromoplasts. How the carotenoid biosynthetic pathway is regulated and what limits total carotenoid accumulation in fruit chromoplasts is not well understood. Here, we have introduced the lycopene β-cyclase genes from the eubacterium Erwinia herbicola and the higher plant daffodil (Narcissus pseudonarcissus) into the tomato plastid genome. While expression of the bacterial enzyme did not strongly alter carotenoid composition, expression of the plant enzyme efficiently converted lycopene, the major storage carotenoid of the tomato fruit, into provitamin A (β-carotene). In green leaves of the transplastomic tomato plants, more lycopene was channeled into the β-branch of carotenoid biosynthesis, resulting in increased accumulation of xanthophyll cycle pigments and correspondingly reduced accumulation of the α-branch xanthophyll lutein. In fruits, most of the lycopene was converted into β-carotene with provitamin A levels reaching 1 mg per g dry weight. Unexpectedly, transplastomic tomatoes also showed a >50% increase in total carotenoid accumulation, indicating that lycopene β-cyclase expression enhanced the flux through the pathway in chromoplasts. Our results provide new insights into the regulation of carotenoid biosynthesis and demonstrate the potential of plastids genome engineering for the nutritional enhancement of food crops.Carotenoids are isoprenoid molecules that are synthesized by all photosynthetic organisms and also by some fungi and nonphotosynthetic bacteria. In plants, they participate in photosynthetic light harvesting and protection against light stress. In addition, carotenoids accumulate to large levels as storage metabolites in chromoplasts of flowers, fruits, and taproots. Carotenoids are also essential to animals, which, however, are unable to synthesize them de novo, and therefore must rely on dietary sources of carotenoids. β-Carotene is the main dietary precursor of vitamin A and therefore also referred to as provitamin A. Vitamin A deficiency in humans represents a global health problem affecting approximately one-third of the countries of the world (Mayer et al., 2008). Presumably due to their antioxidant activity, β-carotene and other carotenoid species also exert protective effects against cardiovascular diseases, certain cancers, and aging-related diseases (Collins, 1999).While the enzymology of the carotenoid biosynthetic pathways in plants and eubacteria is now reasonably well understood (Armstrong, 1997; Cunningham and Gantt, 1998; Hirschberg, 2001), understanding of the regulation of carotenoid biosynthesis is still rather poor (Bramley, 2002). Mainly using the tomato (Solanum lycopersicum) fruit as model system, the study of pigmentation mutants (Ronen et al., 2000; Isaacson et al., 2002; Galpaz et al., 2006) and transgenic approaches (Giuliano et al., 2000, 2008; Römer and Fraser, 2005; Fraser et al., 2007) have provided first insights into regulatory mechanisms operating in carotenogenesis. For example, constitutive expression of the phytoene desaturase (crtI) gene from the bacterium Erwinia uredovora resulted in elevated β-carotene accumulation in tomatoes, but also led to an unexpected reduction in total carotenoid levels (Römer et al., 2000). The reduction in total carotenoids is believed to be an effect of feedback regulation from β-carotene or one of its downstream metabolites (Bramley, 2002). However, fruit-specific overexpression of the native lycopene β-cyclase resulted in increased β-carotene accumulation, without a concomitant decrease in total carotenoids (Rosati et al., 2000). Why some genetic disturbances of carotenoid biosynthesis negatively affect total carotenoid accumulation and others do not (or even result in an increase; Dharmapuri et al., 2002; Fraser et al., 2002), remains to be established.Here we have used tomato plastid transformation to address the regulation of carotenoid biosynthesis exerted at the level of lycopene to β-carotene conversion by the enzyme lycopene β-cyclase (Fig. 1A). We show that plastid expression of a plant lycopene β-cyclase does not only trigger efficient conversion of lycopene to β-carotene, but unexpectedly also results in a >50% increase in total carotenoid accumulation. This contrasts moderately increased β-carotene levels and reduced total carotenoid accumulation upon expression of a bacterial lycopene β-cyclase (Wurbs et al., 2007) and suggests lycopene β-cyclase activity as an important regulatory point in plant and microbial carotenoid biosynthesis.Open in a separate windowFigure 1.Engineering of the carotenoid biosynthetic pathway by plastid transformation. A, Carotenoid biosynthetic pathway in higher plants. The pathway splits into an α-branch and a β-branch immediately downstream of lycopene, the major storage carotenoid of tomato fruits. The enzyme expressed from the tomato plastid genome in this study, lycopene β-cyclase, leads into the β-branch. B, Physical maps of the targeting region in the plastid genome (ptDNA) and the plastid transformation vectors pEcrtY and pNLyc constructed in this study. Genes above the line are transcribed from the left to the right, genes below the line are transcribed in the opposite direction. The transgenes are targeted to the intergenic region between the trnfM and trnfG genes (Ruf et al., 2001). The selectable marker gene aadA is driven by a chimeric rRNA operon promoter (Prrn; Svab and Maliga, 1993), fused to the 3′-UTR from the psbA gene (TpsbA), and flanked by two loxP sites to allow marker removal by Cre-mediated site-specific recombination (Zhou et al., 2008). The transgene expression cassette consists of the ribosomal RNA operon promoter fused to the 5′ leader from the gene 10 of phage T7 (Prrn-G10L; Kuroda and Maliga, 2001) and the 3′-UTR of the rps16 gene (Trps16). Restriction sites used for cloning or RFLP analysis are indicated, and the psaB-derived hybridization probe is denoted by a horizontal bar. Sites lost due to ligation to heterologous ends are in parentheses. C, Southern-blot analysis of tomato transplastomic lines carrying the lycopene β-cyclase gene from daffodil (S.l.-pNLyc) or from E. herbicola (S.l.-pEcrtY). Total cellular DNA was digested with BglII and hybridized to a radioactively labeled probe detecting the psaB region of the plastid genome, which flanks the transgene insertion site (section B). Fragment sizes are given in kb. wt, Wild type. D, Alignment (produced with ClustalW2) of the amino acid sequences of the lycopin β-cyclases from daffodil (Np) and E. herbicola (Eh). Asterisk (*) denotes residues identical in both sequences (marked in bold), colon (:) indicates conserved substitutions, and a dot indicates semiconserved substitutions. The N-terminal extension of the Np sequence is likely to harbor the transit peptide for protein import into plastids. The amino acids that changed due to correction of the Lyc sequence from daffodil (published sequence: GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"X98796.1","term_id":"1419692"}}X98796.1; corrected sequence: accession no. {"type":"entrez-nucleotide","attrs":{"text":"GQ327929","term_id":"254681605"}}GQ327929) are underlined. The corrections improve the sequence similarity in the N-terminal domains of the Np and Eh sequences.     

Dynamics of the actin-binding protein drebrin in motile cells and definition of a juxtanuclear drebrin-enriched zone

The actin-binding protein (ABP) drebrin, isoform E2, is involved in remodelling of the actin cytoskeleton and in formation of cell processes, but its role in cell migration has not yet been investigated. Therefore, we have studied the organization of drebrin in motile cultured cells such as murine B16F1 melanoma and human SV80 fibroblast cells, using live cell confocal microscopy. In cells overexpressing DNA constructs encoding drebrin linked to EGFP, numerous long, branched cell processes were formed which slowly retracted and extended, whereas forward movement was halted. In contrast, stably transfected B16F1 cells containing drebrin-EGFP at physiological levels displayed lamellipodia and were able to migrate on laminin. Surprisingly, in such cells, drebrin was absent from anterior lamellipodia but was enriched in a specific juxtanuclear zone, the "drebrin-enriched zone" (DZ), and in the tail. In leading edges of SV80 cells, characterized by pronounced actin microspikes, drebrin was specifically enriched along posterior portions of the microspikes, together with tropomyosin. Drebrin knock-down by small interfering RNAs did not impair movements of SV80 cells. Our results confirm the role of drebrin E2 in the formation of branching processes and further indicate that during cell migration, the protein contributes to retraction of the cell body and the tail but not to lamellipodia formation. In particular, the novel, sizable juxtanuclear DZ structure will have to be characterized in future experiments with respect to its molecular assembly and cell biological functions.     

Effect of different storage conditions on the mycotoxin contamination of <Emphasis Type=Italic>Fusarium culmorum</Emphasis>-infected and non-infected wheat straw

Mycotoxins are known to affect the health and performance of farm animals. In contrast to cereal grains, the straw is only rarely analysed for mycotoxins, although contaminated straw could additionally expose farm animals to mycotoxins. For this reason, two experiments were carried out to examine the effect of pre-harvest Fusarium infection (inoculation with F. culmorum) and different storage conditions on the mycotoxin concentrations in straw. In the first experiment, both the inoculated and the identically cultivated control straw were stored in rectangular bales either in a barn or outdoors for a time period of 32 weeks (farm-scale experiment). The second experiment was aimed to examine the mycotoxin concentrations during storage under controlled conditions in a temperature-controlled climatic chamber, with target dry matter contents of 86%, 82% and 78% using 1.5-l preservation jars (laboratory-scale experiment). While the concentration of deoxynivalenol and its derivates decreased in the farm-scale experiment when inoculated straw was stored outdoors, the zearalenone concentration increased within the same time period. The latter effect was also detected for the control straw. These opposite effects were probably caused by the massive water uptake during the outdoor storage. The only effect we observed in the laboratory-scale experiment with dry matter contents between 78% and 86% was a more pronounced decrease of the 3-acetyl-deoxynivalenol concentrations in the inoculated straw with increasing moisture contents.     

The non-impact of hunting on moose <Emphasis Type="Italic">Alces alces</Emphasis> movement,diurnal activity,and activity range

Previous studies on moose Alces alces have suggested that interactions with humans may trigger anti-predator behaviors and generate a demographical cost. Therefore, we hypothesized that disturbances from small and big game hunting may have negative effects on moose movements, diurnal activity, and activity range. Using location data from 64 moose equipped with GPS collars from three populations (Low Alpine, Inland, Coastal) with different temporal human presence and spatial accessibility, we evaluated the impact of hunting on moose activity rhythms. On average, female moose in the low human population density (Low Alpine) area (<0.5/km²) had significantly lower movement rates during moose hunting season, but variation in movement rates among individuals were higher compared with female moose in regions with denser human populations (6–24/km²). We found no evidence that reproductive status influenced female moose sensitivity to disturbance. As expected, females used smaller activity ranges and were less active nocturnally than males. The high within-group variation suggests that current hunting disturbance levels do not alter moose population behavior in general. Our data indicate that alterations in movement were related to rutting activity, not human disturbance induced by hunting. In line with behavioral theory, our study suggests that some individuals were more sensitive to hunting disturbance than the general population. Our work suggests that individual moose may perceive human predation risk to be similar to other predation risks.