Transcriptomic analysis of two sheep breeds during lactation, using a new custom microarray platform

We aim at understanding the genomic influence on milk quality and synthesis by comparing two sheep breeds using sheep-specific microarray technology. From sheep ESTs deposited at NCBI we generated a chip carrying about 22,000 non-redundant features in quadruplicate, achieving very good technical outcomes. Oligos were in situ generated on chip using the Combimatrix equipment. We analysed the mammary transcriptome in individuals of two sheep breeds at two lactation stages, to identify genes controlling milk production and metabolic pathways in which these genes are involved. With |FC|>1.4, and p-value≤0.05, 142 and 14 genes resulted differentially expressed in stages 01 and 02, respectively

Cost-Effectiveness Analysis of Dimethyl Fumarate in the Treatment of Relapsing Remitting Multiple Sclerosis: An Italian Societal Perspective

BACKGROUND: Delayed-release dimethyl fumarate (also known as gastro-resistant dimethyl fumarate, hereafter dimethyl fumarate) is an oral disease-modifying therapy used for the treatment of Relapsing-Remitting Multiple Sclerosis (RRMS), an autoimmune chronic inflammatory condition of the central nervous system.OBJECTIVE: The objective of this economic analysis was to compare cost-effectiveness of dimethyl fumarate with the alternatives used as first-line treatment of RRMS in Italy.METHODS: The analysis was conducted from the Italian societal perspective. Health outcomes and costs were evaluated over a 50-year time horizon (equivalent to a lifetime horizon). Both health outcomes and costs were discounted at 3.5%. The cost-effectiveness analysis was conducted by adapting a Markov model, already used in previous similar economic analyses conducted in RRMS, to the Italian context. The Markov model estimated the clinical and economic consequences of treating RRMS patients with the following therapeutic options: dimethyl fumarate; interferon (IFN) beta-1a subcutaneous (SC) at two different doses, 22 mcg and 44 mcg; IFN beta-1b SC; glatiramer acetate (GA) SC 20 mg; oral teriflunomide. Clinical efficacy data were retrieved from an elaboration of an already published mixed treatment comparison (MTC). Both direct and indirect costs (disability, treatment acquisition, administration, monitoring, relapses, adverse events) were included in the analysis. One-way and probabilistic sensitivity analyses were carried out and cost-effectiveness acceptability curves generated.RESULTS: In the base-case analysis, dimethyl fumarate was more efficacious than alternatives, in terms of both survival (19.634 vs. 19.440-19.600 life years for alternatives), and quality-of-life-adjusted survival (6.526 vs. 5.143- 6.189 QALYs for alternatives). The total lifetime cost per patient treated with dimethyl fumarate (€ 954,286) was lower than that of the other DMTs included in the analysis. Therefore, dimethyl fumarate was dominant compared with all analyzed alternatives. Dimethyl fumarate was also the therapeutic option with the highest benefit on disease burden. In fact, costs of disability management were lower than those of all the other first-line drugs included in the analysis. The results of one-way deterministic sensitivity analysis and probabilistic sensitivity analysis confirmed the reliability of base-case results.CONCLUSIONS: The results of the cost-effectiveness analysis confirm that dimethyl fumarate is an optimal first-line treatment for RRMS in Italy, compared with the other first-line alternatives included in the economic analysis, when evaluated from the societal perspective

Hif1α down-regulation is associated with transposition of great arteries in mice treated with a retinoic acid antagonist

<p>Abstract</p> <p>Background</p> <p>Congenital heart defect (CHD) account for 25% of all human congenital abnormalities. However, very few CHD-causing genes have been identified so far. A promising approach for the identification of essential cardiac regulators whose mutations may be linked to human CHD, is the molecular and genetic analysis of heart development. With the use of a triple retinoic acid competitive antagonist (BMS189453) we previously developed a mouse model of congenital heart defects (81%), thymic abnormalities (98%) and neural tube defects (20%). D-TGA (D-transposition of great arteries) was the most prevalent cardiac defect observed (61%). Recently we were able to partially rescue this abnormal phenotype (CHD were reduced to 64.8%, p = 0.05), by oral administration of folic acid (FA). Now we have performed a microarray analysis in our mouse models to discover genes/transcripts potentially implicated in the pathogenesis of this CHD.</p> <p>Results</p> <p>We analysed mouse embryos (8.5 dpc) treated with BMS189453 alone and with BMS189453 plus folic acid (FA) by microarray and qRT-PCR. By selecting a fold change (FC) ≥ ± 1.5, we detected 447 genes that were differentially expressed in BMS-treated embryos vs. untreated control embryos, while 239 genes were differentially expressed in BMS-treated embryos whose mothers had also received FA supplementation vs. BMS-treated embryos. On the basis of microarray and qRT-PCR results, we further analysed the <it>Hif1α </it>gene. In fact <it>Hif1α </it>is down-regulated in BMS-treated embryos vs. untreated controls (FC_micro= -1.79; FC_qRT-PCR= -1.76; p = 0.005) and its expression level is increased in BMS+FA-treated embryos compared to BMS-treated embryos (FC_micro= +1.17; FC_qRT-PCR= +1.28: p = 0.005). Immunofluorescence experiments confirmed the under-expression of Hif1α protein in BMS-treated embryos compared to untreated and BMS+FA-treated embryos and, moreover, we demonstrated that at 8.5 dpc, Hif1α is mainly expressed in the embryo heart region.</p> <p>Conclusions</p> <p>We propose that Hif1α down-regulation in response to blocking retinoic acid binding may contribute to the development of cardiac defects in mouse newborns. In line with our hypothesis, when Hif1α expression level is restored (by supplementation of folic acid), a decrement of CHD is found. To the best of our knowledge, this is the first report that links retinoic acid metabolism to Hif1α regulation and the development of D-TGA.</p

A Tool for Sheep Product Quality: Custom Microarrays from Public Databases

Milk and dairy products are an essential food and an economic resource in many countries. Milk component synthesis and secretion by the mammary gland involve expression of a large number of genes whose nutritional regulation remains poorly defined. The purpose of this study was to gain an understanding of the genomic influence on milk quality and synthesis by comparing two sheep breeds with different milking attitude (Sarda and Gentile di Puglia) using sheep-specific microarray technology. From sheep ESTs deposited at NCBI, we have generated the first annotated microarray developed for sheep with a coverage of most of the genome

Modellistica strutturale e bioinformatica al servizio della ricerca virale: il caso dell'HIV

Nell’ultimo decennio, l’Italia è stata investita da emergenze sanitarie legate alla diffusione planetaria di nuovi ceppi virali, quali la SARS, l’aviaria e la recente influenza A/H1N1. L’approccio combinato sperimentale e computazionale si sta dimostrando sempre più efficace nello studio di queste nuove problematiche, ma anche di malattie virali che sono state oggetto di ricerca nel passato (AIDS, epatite). Una classe di farmaci antiretrovirali assai promettente contro il virus dell’HIV e di recente introduzione è quella che interferisce nel legame del virus con i corecettori presenti sulle cellule target dell’infezione. Gli inibitori del legame al CCR5 risultano attivi solo sui ceppi virali che utilizzano esclusivamente tale corecettore per entrare nelle cellule ospiti. La conoscenza della sequenza genetica dei virus presenti nel singolo paziente può essere utilizzata per valutare a priori se il farmaco antivirale potrà essere efficace nell’inibire questa fase della replicazione del virus

La genomica animale: biodiversità, qualità dei prodotti, salute umana e benessere animale

La genomica studia struttura, funzione ed evoluzione del patrimonio genetico degli esseri viventi, cercando di spiegare come sono programmate le cellule di individui diversi, attraverso l’esplorazione dell’intero patrimonio genetico. Inoltre la genomica studia anche cellule di uno stesso individuo le quali, pur avendo lo stesso genoma, possono avere forma e funzione molto differenti, come ad esempio un neurone ed una cellula di fegato. La comprensione delle complesse relazioni che esistono tra geni ha già portato enormi miglioramenti nella salute umana ed ora sta cominciando ad incidere anche sulla nostra alimentazione ed il benessere degli animali da allevamento. La conoscenza delle sequenze di DNA degli animali allevati può servire sia a migliorare la produzione di latte e carne che la salute stessa degli animali e, di conseguenza, dell’uomo che se ne nutre. Attualmente le analisi di laboratorio consentono di ottenere, in modo accurato e veloce, una grande quantità informazioni sulle sequenze dei geni e dei loro trascritti, ma l’organizzazione dell’enorme quantità di dati prodotti (compito, questo, affidato alla bioinformatica) è, ancora oggi, un “collo di bottiglia” dell’intero processo conoscitivo

Proteomics and Transcriptomics Investigation on longissimus Muscles in Large White and Casertana Pig Breeds

L'articolo è disponibile sul sito dell'editore http://pubs.acs.org/Consumer complaints against the blandness of modern lean meat and the frequent reference to the more strongly flavored meat that was available years ago have prompted reconsideration of high fatdepositing typical pig breeds. Casertana and Large White pig breeds are characterized by a different tendency toward fat accumulation as they exhibit opposite genetic and physiological traits with respect to the energy metabolism. These physiological differences were investigated in longissimus lumborum muscles through proteomics (2-DE, MS/MS) and microarray approaches. Data were analyzed for pathway and network analyses, as well as GO term enrichment of biological functions. As a result, Casertana showed a greater amount of proteins involved in glycolitic metabolism and mainly rely on fast-mobilizable energy sources. Large White overexpressed cell cycle and skeletal muscle growth related genes. Metabolic behavior and other implications are discussed