Coffee species and varietal identification

There are serious economical reasons to pretend warranties in coffee species and varieties authenticity. Arabica adulteration with Robusta coffees, intentional or not, is carried out at different steps of the coffee chain, from plantation to beverage. We present a method based on a real-time PCR technique to perform: a) a qualitative analysis to evaluate the presence/ absence of a species in a sample; b) a quantitative analysis to amplify Robusta samples only, making possible the detection of less than 5% of Robusta in a mixture. Arabica cultivars sold as specialty coffees can be identified and certified by fingerprinting using SSR markers

Serial Analysis of Gene Expression (SAGE) in the Skeletal Muscle of Pig

Skeletal muscle growth represents one of the main economic traits in pig production. To gain a better understanding of expressions profile in pig muscle, serial analysis of gene expression (SAGE) was performed in Longissimus dorsi of two pigs at 3 and 9 months of age. A total of 53,120 long tags were obtained and sequenced from the four muscle SAGE libraries, representing 17,902 different tags, or putative transcripts, 0.64% (+0.09) of which had a relative expression level higher than 1‰. Overall, a total of 218 tags were highly expressed and 31 had a frequency higher than 3‰. Functional characterisation of the expression profiles was performed using Kyoto Encyclopedia of Genes and Genomes metabolic maps and 139 pathways were identified for swine skeletal muscle. Focal adhesion, Mitogen-Activated Protein Kinase signalling, oxidative phosphorylation, ribosomal proteins, regulation of actin cytoskeleton and insulin signalling pathways showed an abundance of genes greater than 1.5% of all the expressed transcripts. A comparison with human SAGE library indicated no statistical differences for the frequency of genes involved in these metabolic pathways

MwoI and SmaI RFLPs polymorphisms of porcine obese gene and their association with carcass and meat characteristics of heavy pigs

The obese gene encodes leptin, a 16-kDa protein involved in the regulation of fat deposition and energy consumption.Backfat is one of the peculiar characteristics of Italian ham, and represents a fundamental quality factor. Therefore, theobese gene can be considered as a candidate marker for determining economically important production traits such asbackfat thickness, feed intake, and growth rate in swine. The aim was to investigate the relationship between obese genepolymorphisms and carcass and meat characteristics of pigs reared for ham production. In the present research, the analysesof three new RFLPs are reported. An MwoI polymorphism occurs at nucleotide 1792, within the intron. Pigs heterozygousat this position have heavier thighs with a thinner layer of fat. Two SmaI polymorphisms occur at nucleotides5018 and 5410 within the 3’ UTR of the obese gene. Animals heterozygous at position 5410 have characteristics suitablefor the production of San Daniele ham: lower backfat thickness and heavier thighs with a thinner fat layer, relative toother genotypes

Endovascular management of massive post-partum haemorrhage in abnormal placental implantation deliveries

Objectives: To retrospectively evaluate safety and efficacy of pelvic artery embolisation (PAE) in post-partum haemorrhage (PPH) in abnormal placental implantation (API) deliveries. Methods: From January 2009 to November 2013, 12 patients with API and intractable intraoperative PPH underwent PAE after caesarean delivery to control a haemorrhage (in four of these cases after hysterectomy). Arterial access was obtained prior to the delivery; PAE was performed in the obstetrics operating room by an interventional radiologist that was present with an interventional radiology (IR) team during the delivery. Results: PAE was successful in preventing bleeding and avoid hysterectomy in four cases (group A). Uterine atony and disseminated intravascular coagulation caused failure of PAE requiring hysterectomy in four patients (group B). PAE prevented bleeding post-hysterectomy in the remaining four cases (group C). Technical success (cessation of contrast extravasation on angiography or occlusion of the selected artery) was 100 %. Maternal and foetal mortality and morbidity were 0 %. Conclusions: PAE is a minimal invasive technique that may help to prevent hysterectomy and control PPH in API pregnancies without complications. Embolisation should be performed on an emergency basis. For such cases, an IR team on standby in the obstetrics theatre may be useful to prevent hysterectomy, blood loss and limit morbidity. Key Points: • Endovascular treatment is a validated technique in post-partum haemorrhage. • Abnormal placental implantation is a risk factor for post-partum haemorrhage. • We propose an interventional radiologist standby in the delivery room. © 2015, European Society of Radiology

Evaluation of gene expression profiles of pig skeletal muscle in response to energy content of the diets using human microarrays

The aim of the research was to compare gene transcription profiles of Musculus longissimus dorsi (MLD) between pigs fed diets with high (HED) or low (LED) energy contents. Two groups of 4 Casertana pigs were reared from 3 to 12 months of age in the same environmental conditions and fed HED or LED. In the HED, the ave rage daily gain and back fat thickness were significantly (P<0.05) higher than in LED pigs. Differential expression of genes in MLD of pigs fed diets with different energy density was assessed by a human high-density complementary DNA (cDNA) muscle microarray consisting of 4670 probes and further confirmed by quantitative real time RT-PCR analysis. Seven of the genes up-regulated in MLD of HED pigs were invo l ved in the glycolytic and ox i d a t i ve metabolism (phosphoglycerate mutase 2, glyc e ra l d e hyde-3-phosphate dehydrogenase, NADH dehydrogenase ubiquinone1 beta 9, muscle pyruvate kinase, enolase 3, muscle creatine kinase, isocitrate dehydrogenase 3 (NAD+) gamma) and four in the contractile apparatus (tropomyosin 1 alpha, troponin C2, fast, fast skeletal myosin light chain 2, troponin T3, skeletal, fast). Instead, HED diet reduced the level of expression of muscle proteins associated with slow fibre type (troponin T1, skeletal, slow, supervillin, myosin binding protein C, slow type, titin, myosin, heavy polypeptide 7, beta, calponin homology-associated smooth muscle) and signal transduction (SH3-binding domain protein 5-like, hypothetical protein FLJ21438, protein kinase cAMP-dependent, catalytic, rho guanine nucleotide exchange factor 15). The down-regulation of CTSB was also observed for HED group. From the results it can be assumed that high energy content of the diet influence physiological processes in the muscle tissue by switching slow fibres into fast reacting fibres and thus enhancing meat quality

GLP-1 Mediates Regulation of Colonic ACE2 Expression by the Bile Acid Receptor GPBAR1 in Inflammation

Background & Aims: ACE2, a carboxypeptidase that generates Ang-(1-7) from Ang II, is highly expressed in the lung, small intestine and colon. GPBAR1, is a G protein bile acid receptor that promotes the release of the insulinotropic factor glucagon-like peptide (GLP)-1 and attenuates intestinal inflammation. Methods: We investigated the expression of ACE2, GLP-1 and GPBAR1 in two cohorts of Crohn’s disease (CD) patients and three mouse models of colitis and Gpbar1−/− mice. Activation of GPBAR1 in these models and in vitro was achieved by BAR501, a selective GPBAR1 agonist. Results: In IBD patients, ACE2 mRNA expression was regulated in a site-specific manner in response to inflammation. While expression of ileal ACE2 mRNA was reduced, the colon expression was induced. Colon expression of ACE2 mRNA in IBD correlated with expression of TNF-α and GPBAR1. A positive correlation occurred between GCG and GPBAR1 in human samples and animal models of colitis. In these models, ACE2 mRNA expression was further upregulated by GPABR1 agonism and reversed by exendin-3, a GLP-1 receptor antagonist. In in vitro studies, liraglutide, a GLP-1 analogue, increased the expression of ACE2 in colon epithelial cells/macrophages co-cultures. Conclusions: ACE2 mRNA expression in the colon of IBD patients and rodent models of colitis is regulated in a TNF-α-and GLP-1-dependent manner. We have identified a GPBAR1/GLP-1 mechanism as a positive modulator of ACE2

A single polyploidization event at the origin of the tetraploid genome of Coffea arabica is responsible for the extremely low genetic variation in wild and cultivated germplasm

The genome of the allotetraploid species Coffea arabica L. was sequenced to assemble independently the two component subgenomes (putatively deriving from C. canephora and C. eugenioides) and to perform a genome-wide analysis of the genetic diversity in cultivated coffee germplasm and in wild populations growing in the center of origin of the species. We assembled a total length of 1.536 Gbp, 444 Mb and 527 Mb of which were assigned to the canephora and eugenioides subgenomes, respectively, and predicted 46,562 gene models, 21,254 and 22,888 of which were assigned to the canephora and to the eugeniodes subgenome, respectively. Through a genome-wide SNP genotyping of 736 C. arabica accessions, we analyzed the genetic diversity in the species and its relationship with geographic distribution and historical records. We observed a weak population structure due to low-frequency derived alleles and highly negative values of Taijma's D, suggesting a recent and severe bottleneck, most likely resulting from a single event of polyploidization, not only for the cultivated germplasm but also for the entire species. This conclusion is strongly supported by forward simulations of mutation accumulation. However, PCA revealed a cline of genetic diversity reflecting a west-to-east geographical distribution from the center of origin in East Africa to the Arabian Peninsula. The extremely low levels of variation observed in the species, as a consequence of the polyploidization event, make the exploitation of diversity within the species for breeding purposes less interesting than in most crop species and stress the need for introgression of new variability from the diploid progenitors