Wood-Specific Polycyclic Aromatic Hydrocarbon (PAH) Patterns in Soot Using Gas Chromatography-Atmospheric Pressure Laser Ionization-Mass Spectrometry (GC-APLI-MS)

<div><p>If organic matter is burnt, the combustion of wood produces the highest amounts of polycyclic aromatic hydrocarbons (PAHs) compared with other fossil energy sources such as oil, coal, or gas. Emissions from wood combustion are increasingly of special interest due to the rising use of wood as a renewable energy source in residential heating in Europe. To the authors' knowledge, reproducible wood-specific PAH patterns in soot were identified for the first time by use of a sampling interval of only 5 min in this study. The short sampling interval was enabled by the very sensitive analytical method of gas chromatography–atmospheric pressure laser ionization–mass spectrometry (GC-APLI-MS) applied. The analysis of 40 PAH of soot from wood logs of spruce, pine, larch (softwood) and beech, birch, oak (hardwood), and wood pellets, as well as wood briquettes, showed 13.46–250.62 mg/kg for ∑40 PAH and 10.75–177.94 mg/kg for the U.S. Environmental Protection Agency PAH standard (without acenaphthylene and anthracene). Highest concentrations occurred in the samples from birch with bark, beech, and wood briquettes. Indeno[1,2,3-<i>cd</i>]pyrene, naphthalene, and alkylated naphthalenes were also detected. Significant concentrations of the very toxic dibenzopyrenes (up to 11.30 mg/kg) are reported. Softwood soot contained highest amounts of 2–4-ring PAH, followed by hardwood which is in accordance with the presence of highest amounts of abietic acid in softwood, a known precursor of retene and phenanthrene. PAH in soot from five spruce samples from different locations show a mean ∑40 PAH concentration of 13.46 mg/kg (<i>n</i> = 5, minimum 8.03, maximum 23.32 mg/kg, SD = 5.65) and exhibited a typical pattern that differed from all other wood soot samples. The distributions of alkylated naphthalenes of the spruce samples show a bell-shape distribution in contrast to the alkylated phenanthrenes/anthracenes of all samples (except the wood pellets), showing a slope distribution. The data indicate that wood-specific PAH patterns exist and under the applied conditions, spruce logs produced the least toxic soot.</p></div

Züchterische Möglichkeiten zur Verminderung der Ebergeruchsproblematik bei Schlachtschweinen

Within the European Union, it can be expected that surgical castration without anaesthesiawill be banned until 2018. Besides immunocastration and surgical castration withanaesthesia, fattening of entire males seems to be an attractive alternative. However, abroad expansion of this practise is problematic due to boar taint which is mainly causedby the pheromone androstenone and bacteria degradation product skatole. Because ofhigh heritabilities of both components, breeding against boar taint should be successful.Due to lack of certain thresholds for androstenone, a precise definition of the breeding objective is difficult. Human nose scoring is less influenced by the problem of threshold determination. However, in order to integrate the results of human nose scoring into the breeding procedure, it is necessary to define clear guidelines for recording. Breeding against androstenone is impaired by the unfavourable correlation particular to maternal fertility. Although this antagonism is not obvious in some commercial populations, only selection within sire lines is feasible, whereas breeding against androstenone in dam lines is not recommendable. Molecular genetic tools enable to identify genes, involved only in the degradation, but not in the synthesis of androstenone. In other words, there are no pleiotropic effects of these genes on androstenone and fertility. Recent studies confirm the existence of such non-pleiotropic genes. Therefore selection against androstenone could be applied efficiently without negative consequences on maternal fertility

The Effect of Protein Abundance in Porcine Skeletal Muscle on the Expression of Water Holding Capacity

Water holding capacity measured as drip loss is the ability of the skeletal muscle to retain water post-mortem. The aim of this work was a global proteome analysis in order to identify the relevant biological mechanism of porcine muscle proteins, with potential functional relevance for drip loss. For this experiment a Duroc x Pietrain F2 resource population (n = 42) was used. Proteins were isolated from the Musculus longissimus dorsi of the F2 animals. The relative protein quantification was performed using isotope-coded protein labeling techniques and electrospray ionization liquid-chromatography-tandem mass spectrometry. In total, 763 different proteins were identified. The proteins, PYGL, PYGM, HSPA8, EE1A1, ACTA1, CASQ1, FLN-C, MYOM1, TNNT3, &nbsp;and HSP27 were up regulated and TNNI1, MYL3, MYL2, MB, MYBPC1, FHL1C, TPM1, TPM2, AK1, TNNC2, MYL11, CK, PGK1 and MYH7were down-regulated comparing animals with low and high drip loss. Results revealed that the high drip loss meat was characterized &nbsp;by higher level of glycolytic enzymes than in low drip loss meat. Additionally, we could observe that higher levels of chaperone proteins were associated with a low drip loss level.</p

Integrative Analysis of Metabolomic, Proteomic and Genomic Data to Reveal Functional Pathways and Candidate Genes for Drip Loss in Pigs

The aim of this study was to integrate multi omics data to characterize underlying functional pathways and candidate genes for drip loss in pigs. The consideration of different omics levels allows elucidating the black box of phenotype expression. Metabolite and protein profiling was applied in Musculus longissimus dorsi samples of 97 Duroc &times; Pietrain pigs. In total, 126 and 35 annotated metabolites and proteins were quantified, respectively. In addition, all animals were genotyped with the porcine 60 k Illumina beadchip. An enrichment analysis resulted in 10 pathways, amongst others, sphingolipid metabolism and glycolysis/gluconeogenesis, with significant influence on drip loss. Drip loss and 22 metabolic components were analyzed as intermediate phenotypes within a genome-wide association study (GWAS). We detected significantly associated genetic markers and candidate genes for drip loss and for most of the metabolic components. On chromosome 18, a region with promising candidate genes was identified based on SNPs associated with drip loss, the protein &quot;phosphoglycerate mutase 2&quot; and the metabolite glycine. We hypothesize that association studies based on intermediate phenotypes are able to provide comprehensive insights in the genetic variation of genes directly involved in the metabolism of performance traits. In this way, the analyses contribute to identify reliable candidate genes.</p

Genomic analysis of water holding capacity of meat in a porcine resource population

In this study lean meat water-holding capacity (WHC) of a Duroc &times; Pietrain (DuPi) resource populations withcorresponding genotypes and transcriptomes were investigated using the approaches of genetical genomics.WHC was characterized by drip loss measured in M. long. dorsi. Genotypes of 169 F2 DuPi animals wereidentified by the 60k Illumina SNP chips. Whole genome tanscriptomes of muscle were available for 132F2 animals using the 24k Affymetrix expression microarray. Performing genome wide associations studies(GWAS) of transcriptional profiles which are correlated with phenotypes allows elucidating cis- and trans-regulation. Expression levels of 1,228 genes were significantly correlated with drip loss and were furtheranalyzed for enrichment of functional annotation groups as defined by gene ontology and KEGG pathways.A hyper geometric gene set enrichment test was performed and revealed glycolysis/glyconeogenesis, pentosephosphate pathway and pyruvat metabolism as most promising pathways. For 267 selected transcripts eQTLanalysis was performed and revealed 78,576 significant (P&lt;0.01) associations in total. Because of positionalaccordance of the gene underlying transcript and the eQTL location it was possible to identify 59 eQTL thatcan be assumed to be cis-regulated. Comparing the results of gene set enrichment and the eQTL detectiontests molecular networks and potential candidate genes which seem to play key roles in the expression ofwater holding capacity were detected. One gene was the glucose-6-phosphate transporter SLC37A4 whichwas assumed to be cis-regulated and involved in the glucose metabolic process. This approach supports toidentify trait-associated SNPs and to understand the biology of complex traits.</div

Association study and expression analysis of CD9 as candidate gene for boar sperm quality and fertility traits

Cluster-of-differentiation antigen 9 (CD9) gene expressed in the male germ line stem cells is crucial for sperm-egg fusion, and was therefore selected as candidate gene for boar semen quality. The association of CD9 with boar sperm quality and fertility trait was analyzed using a total of 340 boars both from purebred Pietrain and Pietrain x Hampshire crosses. A single nucleotide polymorphism (g.358A>T) in intron 6 was significantly associated with sperm motility (MOT) (P<0.001), plasma droplet rate (PDR) (P<0.001) and abnormal spermatozoa rate (ASR) (P<0.01).Boars were divided into two groups with group 1 (G-I) boars having a higher SCON and SMOT, lower SVOL (sperm volume) and group 2 (G-II) having a lower SCON and SMOT, higher SVOL The mRNA and protein expression levels were evaluated in reproductive, non-reproductive tissues and spermatozoa from G-I and G-II animals by using quantitative real-time PCR and western blotting. When both reproductive and nonreproductive tissues were examined, highest mRNA was expressed in prostate gland, then in the body of the epididymis, vas deferens and tail of the epididymis. In case of reproductive tissues, CD9 expression was higher in tissues and spermatozoa collected from G-I boars than those collected from G-II boars. The mRNA expression was significantly different (P<0.05) in body of epididymis from G-I and G-II boars. The CD9 protein expression results from western blot were coincided with the results of qRT-PCR. Moreover, CD9 protein localization in Leydig cells, Sertoli cells, epithelial cells and spermatozoa was remarkable which indicated the important role of CD9 in spermatogenesis process. By using mRNA and protein expression profiles, it could be shown that CD9 plays a crucial role during sperm development, especially within the epididymis where the maturation of the sperm, a key process for the sperm quality and motility takes place. These results will improve the understanding of the functions of the CD9 in spermatogenesis within the reproductive tracts and will shed light on CD9 as a candidate gene in the selection of good sperm quality boars. (C) 2011 Elsevier B.V. All rights reserved

Sulforaphane enhances proliferation ofporcine satellite cells through suppression ofTGF-beta signaling pathway

Satellite cells, the muscle stem cells, play a critical role in muscle growth, maintenance, and regeneration. A lot of muscle diseases result from defective function of satellite cells. Porcine satellite cells are a good model for studying the role of satellite cells in muscle development. Sulforaphane (SFN), a natural molecule rich in cruciferous vegetables, is a potent inducer for the NF-E2-related factor 2 (Nrf2) signaling and also inhibits the activity of histone deacetylases (HDAC). Our previous study found that SFN epigenetically suppressed the transcription of myostatin in porcine satellite cells. However, the effects of SFN on the proliferation of porcine satellite cells and the related mechanisms are far from understood. In the present study, we report that SFN enhanced the proliferation of the porcine satellite cells and modified the expression myogenic regulatory factors. SFN altered the expression of HDACs and inhibited the activity of HDACs. The activity of TGF-? signaling was suppressed by SFN treatment, which was accompanied with up-regulated Smad7, an endogenous suppressor of TGF-? signaling. Furthermore, we found that SFN increased the mRNA expression of Smad7&rsquo;s transcription factors and decreased the expression of miRNAs targeting Smad7. The DNA methylation of a studied fragment in Smad7 promoter was not influenced by SFN treatment. SFN has received substantial attention because of its potential application in cancer therapy. The present study, for&nbsp;the first time, investigated the effects of SFN on the proliferation of porcine satellite cells and the underlying mechanism. We found that both mRNA and protein level of Smad7 were greatly increased by SFN. Thus, besides reducing TGF-?1 protein abundance, SFN also inhibits the activity of TGF-? signaling by increasing expression of Smad7. It has been shown that overexpression of Smad7 led to enhanced skeletal muscle differentiation and cellular hypertrophy. In summary, our studies state that SFN enhances the proliferation of porcine satellite cells by suppressing TGF-? signaling through activation of Smad7.</p

Identification of gene co-expression clusters in liver tissues from multiple porcine populations with high and low backfat androstenone phenotype

Background: Boar taint is principally caused by accumulation of androstenone and skatole in adipose tissues. Studies have shown high heritability estimates for androstenone whereas skatole production is mainly dependent on nutritional factors. Androstenone is a lipophilic steroid mainly metabolized in liver. Majority of the studies on hepatic androstenone metabolism focus only on a single breed and very few studies account for population similarities/differences in gene expression patterns. In this work, we concentrated on population similarities in gene expression to identify the common genes involved in hepatic androstenone metabolism of multiple pig populations. Based on androstenone measurements, publicly available gene expression datasets from three porcine populations were compiled into either low or high androstenone dataset. Gene expression correlation coefficients from these datasets were converted to rank ratios and joint probabilities of these rank ratios were used to generate dataset specific co-expression clusters. Finally, these networks were clustered using a graph clustering technique.BackgroundBoar taint is principally caused by accumulation of androstenone and skatole in adipose tissues. Studies have shown high heritability estimates for androstenone whereas skatole production is mainly dependent on nutritional factors. Androstenone is a lipophilic steroid mainly metabolized in liver. Majority of the studies on hepatic androstenone metabolism focus only on a single breed and very few studies account for population similarities/differences in gene expression patterns. In this work, we concentrated on population similarities in gene expression to identify the common genes involved in hepatic androstenone metabolism of multiple pig populations. Based on androstenone measurements, publicly available gene expression datasets from three porcine populations were compiled into either low or high androstenone dataset. Gene expression correlation coefficients from these datasets were converted to rank ratios and joint probabilities of these rank ratios were used to generate dataset specific co-expression clusters. Finally, these networks were clustered using a graph clustering technique.ResultsCluster analysis identified a number of statistically significant co-expression clusters in the dataset. Further enrichment analysis of these clusters showed that one of the clusters from low androstenone dataset was highly enriched for xenobiotic, drug, cholesterol and lipid metabolism and cytochrome P450 associated metabolism of drugs and xenobiotics. Literature references revealed that a number of genes in this cluster were involved in phase I and phase II metabolism. Physical and functional similarity assessment showed that the members of this cluster were dispersed across multiple clusters in high androstenone dataset, possibly indicating a weak co-expression of these genes in high androstenone dataset.ConclusionsBased on these results we hypothesize that majority of the genes in this cluster forms a signature co-expression cluster in low androstenone dataset in our experiment and that majority of the members of this cluster might be responsible for hepatic androstenone metabolism across all the three populations used in our study. We propose these results as a background work towards understanding breed similarities in hepatic androstenone metabolism. Additional large scale experiments using data from multiple porcine breeds are necessary to validate these findings.</p

A Genome-Wide Association Study in Large White and Landrace Pig Populations for Number Piglets Born Alive

<div><p>The number of piglets born alive (NBA) per litter is one of the most important traits in pig breeding due to its influence on production efficiency. It is difficult to improve NBA because the heritability of the trait is low and it is governed by a high number of loci with low to moderate effects. To clarify the biological and genetic background of NBA, genome-wide association studies (GWAS) were performed using 4,012 Large White and Landrace pigs from herdbook and commercial breeding companies in Germany (3), Austria (1) and Switzerland (1). The animals were genotyped with the Illumina PorcineSNP60 BeadChip. Because of population stratifications within and between breeds, clusters were formed using the genetic distances between the populations. Five clusters for each breed were formed and analysed by GWAS approaches. In total, 17 different significant markers affecting NBA were found in regions with known effects on female reproduction. No overlapping significant chromosome areas or QTL between Large White and Landrace breed were detected.</p></div