Quantitative real time PCR approach to study gene expression profile during prenatal growth of skeletal muscle in pig of Duroc and Pietrain breeds

The quantitative real time-PCR (QRT-PCR) is a very sensitive method used to quantify mRNA level in gene expression analysis. Combining amplification, detection and quantification in a single step, allows a more accurate measurement compared to the traditional PCR end point analysis (Pfaffl, 2001; Bustin, 2002)

Identification of SNPs in the promoter of \u3b2-lactoglobulin gene in three Sicilian goat breeds

The aim of this work was to sequence the full-length promoter region of the caprine \u3b2-lactoglobulin (\u3b2-lg) gene in three Sicilian goat breeds (Girgentana, Maltese, and Derivata di Siria), in order to identify polymorphisms, to search for transcription factors (TFs) sites, and to check if polymorphisms found lay within TFs binding sites. The promoter region of \u3b2-lg gene in Sicilian goat breeds showed high level of polymorphism due to the presence of 31 SNPs. Binding sites for several TFs were found within the goat \u3b2-lg promoter and within regions conserved between ovine and caprine species. Two SNPs were detected within TFs binding sites, such as MPBF and NF-I. Further studies are in progress to confirm polymorphic sites, to evaluate the possible effect of these mutations on binding affinity of TFs, their relationship with \u3b2-lg gene expression, and the functional role of SNPs within the TFs sites of the promoter region on milk trait

Polymorphisms of beta-lactoglobulin promoter region in three Sicilian goat breeds

Several beta-lactoglobulin (BLG) polymorphisms have been described within the proximal promoter region and coding region of the caprine gene, although no genetic variants affecting the protein amino acid composition and/or expression level have been characterized so far. Binding sites for several transcription factors (TFs) are present in the BLG promoter region. The aims of this work were to sequence the full-length promoter region of three Sicilian goat breeds in order to identify polymorphisms, analyze the identified haplotypes, search for differences between breeds for the presence of polymorphisms in this gene region, search for putative TFs binding sites, and check if polymorphisms lay within the identified TFs binding sites. The promoter region of BLG gene in Sicilian goat breeds showed high level of polymorphism due to the presence of 36 single nucleotide polymorphisms (SNPs). Association between polymorphic sites was computed within the whole sample analyzed and 18 haplotypes were inferred. Binding sites for three milk protein binding factors (MPBFs) and four nuclear factor-I (NF-I) were found within BLG promoter region based on the ovine sequence. The identification of some SNPs within TFs binding sites allowed hypothesizing the loss of TFs. Further studies are in progress to evaluate the effect of these mutations on binding affinity of TFs, the functional interaction of the TFs with the goat BLG promoter, and the relationship of the polymorphisms with BLG gene expression and milk production and composition

skeletal muscle expression analysis of fat metabolism genes in pig

Fat content and backfat thickness are important polygenic parameters influencing meat quality and carcass traits in pigs. Up to now, there is a lack of knowledge on the expression level of the genes encoding for enzymes involved in fatty acid metabolism in porcine skeletal muscle. In the present study we analysed, by quantitative real time PCR, the expression of three genes, acetyl-CoA carboxylase (ACACA), ATP citrate lyase (ACL) and fatty acid synthase (FASN) in skeletal muscle tissue samples of Italian Large White and Italian Duroc pigs with divergent breeding values for backfat thickness or visible intramuscular fat. Significant differences of the expression level for ACACA gene (P=0.04) and for ACL gene (P=0.02) were observed between the two breeds, comparing the samples selected for backfat thickness trait. The expression analysis of FASN gene in the samples with different genotype at the SNP c.265C>T showed that the TT genotype presented the lowest values in both breeds. The differences observed between breeds should be further considered to investigate the putative involvement of these genes on fat deposition traits. - Ricerca e analisi di espressione di geni candidati per la deposizione di grasso nella carne suina in razze con diversa attitudine alla deposizione di grasso intramuscolare

Deregulated PTEN/PI3K/AKT/mTOR signaling in prostate cancer: Still a potential druggable target?

Although the prognosis of patients with localized prostate cancer is good after surgery, with a favorable response to androgen deprivation therapy, about one third of them invariably relapse, and progress to castration-resistant prostate cancer. Overall, prostate cancer therapies remain scarcely effective, thus it is mandatory to devise alternative treatments enhancing the efficacy of surgical castration and hormone administration. Dysregulation of the phosphoinositide 3-kinase pathway has attracted growing attention in prostate cancer due to the highly frequent association of epigenetic and post-translational modifications as well as to genetic alterations of both phosphoinositide 3-kinase and PTEN to onset and/or progression of this malignancy, and to resistance to canonical androgen-deprivation therapy. Here we provide a summary of the biological functions of the major players of this cascade and their deregulation in prostate cancer, summarizing the results of preclinical and clinical studies with PI3K signaling inhibitors and the reasons of failure independent from genomic changes

Investigation of SNPs in the ATP1A2, CA3 and DECR1 genes mapped to porcine chromosome 4: analysis in groups of pigs divergent for meat production and quality traits

STUDIO DI TRE GENI (ATP1A2, CA3E DECR1) LOCALIZZATI SUL CROMOSOMA SUINO 4: ANALISIDELLE FREQUENZE ALLELICHE DI SNP IN SUINI ESTREMI PER ALCUNI CARATTERI PRODUTTIVITre geni (ATPase, Na+/K+transporting, \u3b12(+) polypeptide, ATP1A2; carbonic anhydrase III, CA3; 2,4-die-noyl CoA reductase 1, mitochondrial, DECR1), isolati da una libreria a cDNA ottenuta da muscolo scheletri-co di suino, sono stati scelti per questo studio sulla base del ruolo fisiologico della proteina codificata in pro-cessi cellulari e metabolici collegabili in modo diretto o indiretto con alcuni caratteri produttivi. La scelta dei tre geni \ue8 stata anche basata sulla loro localizzazione sul cromosoma 4 dove diversi QTL per caratteri pro-duttivi sono gi\ue0 stati identificati.Nella regione 3\u2019 non tradotta del gene CA3\ue8 stato identificato, mediante analisi SSCP, un polimorfismo bial-lelico. Il sequenziamento dei due alleli ha permesso di identificare una nuova mutazione puntiforme, che \ue8stata successivamente analizzata mediante PCR-RFLP. Per questo gene \ue8 stato effettuato il mappaggiogenetico sul cromosoma 4 mediante l\u2019analisi della mutazione nei campioni delle famiglie di riferimento delprogetto europeo di mappaggio del genoma suino (PiGMaP). Utilizzando il polimorfismo PCR-RFLP del geneATP1A2, gi\ue0 decritto in un precedente lavoro come SSCP, sono state tipizzate diverse famiglie di riferimen-to PiGMaP ed \ue8 stato confermato il mappaggio genetico di ATP1A2. Utilizzando queste informazioni e quel-le gi\ue0 disponibili per DECR1, per la prima volta, \ue8 stata ottenuta una mappa di linkage del cromosoma 4che comprende tutti e tre i geni analizzati. Il mappaggio genetico dei tre geni \ue8 stato anche confermatomediante tipizzazione di un pannello di ibridi di cellule irradiate (IMpRH 7000 rad).Le frequenze alleliche delle mutazioni identificate nei tre loci sono state studiate in 11 razze suine (LargeWhite Italiana, Landrace Italiana, Duroc Italiana, Landrace Belga, Hampshire, Pi\ue9train, Meishan, CintaSenese, Casertana, Calabrese and Nero Siciliano) per un totale di 272 animali. Inoltre, come approccio ini-ziale per poi scegliere i geni da analizzare in futuri studi di associazione, abbiamo confrontato le frequenzealleliche di mutazioni puntiformi per questi tre loci in gruppi di suini di razza Large White Italiana e DurocItaliana, analizzando animali con valori degli indici genetici estremi e divergenti per alcuni caratteri produt-tivi (accrescimento, spessore lardo dorsale, tagli magri e grasso intermuscolare visibile). Per il gene CA3 \ue8stata osservata una differenza nella distribuzione delle frequenze alleliche (P< 0,05) per i due caratteri taglimagri (nella razza Large White Italiana) e grasso intermusculare visibile (nella razza Duroc Italiana). Per ilgene DECR1, differenze significative sono state osservate per il carattere grasso intermuscolare visibile. Ilgene ATP1A2, che mappa vicino al locus FAT1, non ha presentato differenze statisticamente significativenelle frequenze tra i gruppi estremi per i caratteri oggetto di studio. Analizzando il livello di linkage disequi-librium(LD) tra i tre loci, \ue8 stato evidenziato un elevato livello di LD (D\u2019= 0,967; P< 0,0001) tra i geni CA3e DECR1, solo nella popolazione Duroc Italiana. Questi primi risultati pongono le basi per ulteriori studi perverificare se i geni CA3e DECR1sono associati con i caratteri oggetto di selezione nel suino pesante.Three genes (ATPase, Na+/K+ transporting, \u3b1 2(+) polypeptide, ATP1A2; carbonic anhydrase III, CA3; 2,4-dienoyl CoA reductase 1, mitochondrial, DECR1), isolated from a porcine skeletal muscle cDNA library and mapped on porcine chromosome 4 (SSC4), were investigated. A new single nucleotide polymorphism (SNP) was identified in the 3\u2019-untranslated region of the CA3 gene and used to genetically map this locus on SSC4 together with the ATP1A2 and DECR1 loci for which SNPs were already reported. Allele frequencies of the three loci were reported for 11 pig breeds (Italian Large White, Italian Landrace, Italian Duroc, Belgian Landrace, Hampshire, Pi\uf9train, Meishan, Cinta Senese, Casertana, Calabrese and Nero Siciliano). Radiation hybrid mapping of these genes confirmed the linkage mapping results as well as mapping information reported by other authors. Then, the SNPs identified in the ATP1A2, CA3 and DECR1 genes were genotyped in Italian Large White and Italian Duroc animal groups with extreme and divergent estimated breeding value for several production traits. For CA3 significant differences in allele frequencies (P< 0.05) were observed between the extreme groups of pigs for the lean cuts (Italian Large White) and visible intermuscular fat (Italian Duroc) traits. For DECR1, a significant difference in allele frequencies was observed only for the visible intermuscular fat trait. ATP1A2, which maps close to the FAT1 locus, did not show any significant difference. A very high linkage disequilibrium (D\u2019= 0.967; P< 0.0001) was identified between CA3 and DECR1 in the Italian Duroc population. Further investigations are needed to evaluate the effect of CA3 and DECR1 on the considered trait

Exploiting the Potential in Water Cleanup from Metals and Nutrients of Desmodesmus sp. and Ampelodesmos mauritanicus

Increasing levels of freshwater contaminants, mainly due to anthropogenic activities, have resulted in a great deal of interest in finding new eco-friendly, cost-effective and efficient methods for remediating polluted waters. The aim of this work was to assess the feasibility of using a green microalga Desmodesmus sp., a cyanobacterium Nostoc sp. and a hemicryptophyte Ampelodesmos mauritanicus to bioremediate a water polluted with an excess of nutrients (nitrogen and phosphorus) and heavy metals (copper and nickel). We immediately determined that Nostoc sp. was sensitive to metal toxicity, and thus Desmodesmus sp. was chosen for sequential tests with A. mauritanicus. First, A. mauritanicus plants were grown in the ‘polluted’ culture medium for seven days and were, then, substituted by Desmodesmus sp. for a further seven days (14 days in total). Heavy metals were shown to negatively affect both the growth rates and nutrient removal capacity. The sequential approach resulted in high metal removal rates in the single metal solutions up to 74% for Cu and 85% for Ni, while, in the bi-metal solutions, the removal rates were lower and showed a bias for Cu uptake. Single species controls showed better outcomes; however, further studies are necessary to investigate the behavior of new specie

Two fossil groups of galaxies at z~0.4 in the COSMOS: accelerated stellar-mass build-up, different progenitors

We report on 2 fossil groups of galaxies at z=0.425 and 0.372 discovered in the Cosmic Evolution Survey (COSMOS) area. Selected as X-ray extended sources, they have total masses (M_200) of 1.9(+/-0.41)E13 and 9.5(+/-0.42)E13 M_sun, respectively, as obtained from a recent X-ray luminosity-mass scaling relation. The lower mass system appears isolated, whereas the other sits in a well-known large-scale structure (LSS) populated by 27 other X-ray emitting groups. The identification as fossil is based on the i-band photometry of all the galaxies with a photo-z consistent with that of the group at the 2-sigma confidence level and within a projected group-centric distance equal to 0.5R_200, and i_AB<=22.5-mag limited spectroscopy. Both fossil groups exhibit high stellar-to-total mass ratios compared to all the X-ray selected groups of similar mass at 0.3<=z<=0.5 in the COSMOS. At variance with the composite galaxy stellar mass functions (GSMFs) of similarly massive systems, both fossil group GSMFs are dominated by passively evolving galaxies down to M^stars~1E10 M_sun (according to the galaxy broad-band spectral energy distributions). The relative lack of star-forming galaxies with 1E10<=M^stars<=1E11 M_sun is confirmed by the galaxy distribution in the b-r vs i color-magnitude diagram. Hence, the 2 fossil groups appear as more mature than the coeval, similarly massive groups. Their overall star formation activity ended rapidly after an accelerated build up of the total stellar mass; no significant infall of galaxies with M^stars>=1E10 M_sun took place in the last 3 to 6 Gyr. This similarity holds although the 2 fossil groups are embedded in two very different density environments of the LSS, which suggests that their galaxy populations were shaped by processes that do not depend on the LSS. However, their progenitors may do so. ...Comment: 12 pages, 5 color figures, 1 table; to be published in the MNRA

BLAST Observations of the South Ecliptic Pole field: Number Counts and Source Catalogs

We present results from a survey carried out by the Balloon-borne Large Aperture Submillimeter Telescope (BLAST) on a 9 deg^2 field near the South Ecliptic Pole at 250, 350 and 500 {\mu}m. The median 1{\sigma} depths of the maps are 36.0, 26.4 and 18.4 mJy, respectively. We apply a statistical method to estimate submillimeter galaxy number counts and find that they are in agreement with other measurements made with the same instrument and with the more recent results from Herschel/SPIRE. Thanks to the large field observed, the new measurements give additional constraints on the bright end of the counts. We identify 132, 89 and 61 sources with S/N>4 at 250, 350, 500 {\mu}m, respectively and provide a multi-wavelength combined catalog of 232 sources with a significance >4{\sigma} in at least one BLAST band. The new BLAST maps and catalogs are available publicly at http://blastexperiment.info.Comment: 25 pages, 6 figures, 4 tables, Accepted by ApJS. Maps and catalogs available at http://blastexperiment.info