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)

A critical review of the mechanisms involved in the occurrence of growth-related abnormalities affecting broiler chicken breast muscles

In the past decade, the poultry industry has faced the occurrence of growth-related muscular abnormalities that mainly affect, with a high incidence rate, the Pectoralis major of the fast-growing genotypes selected for their production performances (high growth rate and breast yield). These myopathies are termed as White Striping, Wooden Breast, and Spaghetti Meat and exhibit distinctive phenotypes. A spatiotemporal distribution has been demonstrated for these disorders as in the early stage they primarily affect the superficial area in the cranial portion of the muscle and, as the birds grow older, involve the entire tissue. Aside from their distinctive phenotypes, these myopathies share common histological features. Thus, it might be speculated that common causative mechanisms might be responsible for the physiological and structural perturbations in the muscle associated with these conditions and might underpin their occurrence. The present review paper aims to represent a critical survey of the outcomes of all the histologic and ultrastructural observations carried out on White Striping, Wooden Breast, and Spaghetti Meat affected muscles. Our analysis has been performed by combining these outcomes with the findings of the genetic studies, trying to identify possible initial causative mechanisms triggering the onset and the time-series of the events ultimately resulting in the development and progression of the growth-related myopathies currently affecting broilers Pectoralis major muscles. Several evidences support the hypothesis that sarcoplasmic reticulum stress, primarily induced an accumulation of misfolded proteins (but also driven by other factors including altered calcium homeostasis and accumulation of fatty acids), may be responsible for the onset of these growth-related myopathies in broilers. At the same time, the development of hypoxic conditions, as a direct consequence of an inadequate vascularization, triggers a time-series sequence of events (i.e., phlebitis, oxidative stress, etc.) resulting in the activation of response mechanisms (i.e., modifications in the energetic metabolism, inflammation, degeneration, and regeneration) which are all strictly related to the progression of these myopathic disorders

Beta-lactoglobulin polymorphism in Girgentana goat breed

Beta-lactoglobulin (b-lg) is a globular protein belonging to the lipocalin family. It is the major whey protein in the milk of ruminants. It is also present in the milk of most mammals but is lacking in rodents, lagomorphs and humans. A large number of variants have been reported for cow and sheep milk. Several studies have shown association between b-lg variants and milk production and composition, even if the results are not always concordant. In goat, no b-lg variants related with amino acid change have been characterized at DNA level, but some authors described the presence of polymorphisms in the 3’UTR and in the proximal promoter region. Mutations in the promoter region could be those most likely responsible for different level of gene expression. The aim of this work was to study the genetic polymorphism at DNA level of b-lg gene in Girgentana goat breed. A total of 238 genomic DNA samples of Girgentana breed were genotyped. A fragment of 709 bp, including 588 bp of proximal promoter region and 121 bp of exon 1, was amplified using primers GOAPF3 and GoatE1R2. PCR-RFLP procedure was used for fast detection of two single nucleotide substitutions as described by Graziano et al. (2003). The base substitutions originating the polymorphic sites consist of: 1. a transition T›C at position -341 and 2. a transition C›T at position -60. A FspBI PCR-RFLP protocol was used to detect the mutation -341 (T/C) and a SmaI PCR-RFLP protocol for the mutation -60 (C/T) of the proximal promoter region. The allelic frequencies and the Hardy-Weinberg equilibrium were estimated using the GENEPOP software. Girgentana goat breed shows no significant deviation from Hardy- Weinberg equilibrium for the allele frequencies found in both polymorphic sites considered. The genotypic frequencies for both mutations resulted in 0.65 (T/T), 0.33 (T/C) and 0.02 (C/C) for the position -341, and 0.82 (C/C), 0.17 (C/T) and 0.01 (T/T) for the position -60. These results are in agreement with the previous obtained by Graziano et al. (2003) in the same breed. Further analysis are in progress to investigate the possible effect of these variants on the expression of b-lg gene, on the milk protein composition and on milk production traits

Multi-site Resource Allocation in a QoS-Aware 5G Infrastructure

Network softwarization has paved the way for 5G technologies, and a wide-range of (radically new) verticals. As the telecommunications infrastructure evolves into a sort of distributed datacenter, multiple tenants such as vertical industries and network service providers share its aggregate pool of resources (e.g., networking, computing, etc.) in a layered \u201das-a-Service\u201d approach exposed as slice abstractions. The challenge remains in the coordination of various stakeholders\u2019 assets in realizing end-to-end network slices and supporting the multi-site deployment and chaining of the micro-service components needed to implement cloud-native vertical applications (vApps). In this context, particular care must be taken to ensure that the required resources are identified, made available and managed in a way that satisfies the vApp requirements, allows for a fair share of resources and has a reasonable impact on the overall vApp deployment time. With these challenges in mind, this paper presents the Resource Selection Optimizer (RSO)-a software-service in the MATILDA Operations Support System (OSS), whose main goal is to select the most appropriate network and computing resources (according to some criterion) among a list of options provided by the Wide-area Infrastructure Manager (WIM). It consists of three submodules that respectively handle: (i) the aggregation of vApp components based on affinities, (ii) the forecasting of (micro-) datacenter resources utilization, (iii) and the multi-site placement of the (aggregated) vApp micro-service components. The RSO\u2019s performance is mainly evaluated in terms of the execution times of its submodules while varying their respective input parameters, and additionally, three selection policies are also compared. Experimental results aim to highlight the RSO behavior in both execution times and deployment costs, as well as the RSO interactions with other OSS submodules and network platform components, not only for multi-site vApp deployment but also for other network/services management operations

Genome-wide association study identifies markers associated with carcass and meat quality traits in Italian Large White pigs

A GWAS was performed using the genotypes obtained by PorcineSNP60 v2 BeadChip and 11 phenotypic traits (carcass lean meat percentage; backfat thickness; Longissimus thoracis muscle thickness; lightness; backfat thickness measured with caliper at the midline; meat pH measured at about 1 h post mortem and 24 h post mortem; CIE L*, a* and b* color parameters; and water-holding capacity). Three markers were associated with three of the phenotypic traits considered: M1GA0009592 (SSC7) with backfat thickness and lean meat content, DIAS0002910 (SSC6) and ALGA0109856 (SSC6) with water-holding capacity. The marker M1GA0009592, associated with backfat thickness, lies in a QTL region near the gene JARID2, which is a transcription factor also involved in the regulation of adipose-derived stem cell pluripotency. The results seem to indicate a possible role of these genomic regions in the regulation of pig carcass fatness (i.e. backfat at last rib) and water-holding capacity

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

Analysis of the melanocortin receptor 1 (MC1R) gene in Sicilian goat breeds

AbstractMammalian coat colour is mainly determined by the distribution of two different types of melanins: pheomelanin (red/yellow pigments) and eumelanin (black pigments). Their synthesis is regulated by the melanocortin 1 receptor (MC1R/Extension locus) that binds the α-melanocyte-stimulating hormone (α-MSH) and the agouti signalling protein (ASIP, coded by the Agouti locus). In mammals, several studies have reported that loss-of-function mutations in MC1R lead to red/yellow pigmentation, while gain-of-function mutations lead to black/dark colours. Mutations at the Agouti locus exert, in general, epistatic interactions on the Extension locus. In goats, classical genetic studies have indicated that variations at the Agouti locus may be the main source of colour variability within and between breeds, while the effect of the Extension locus on this phenotypic trait has been only partially deduced. In order to better understand the role of the Extension locus on coat colour in this species, here we analysed..

Research Notes : United States : A greenhouse method of screening soybeans for resistance to Fusarium wilt

Fusarium wilt of soybean (causal organism: Fusarium oxysporum Schlecht. emend. Snyd. & Hans.) has become an increasingly severe disease in the breed-ing plots at Gainesville and may be an undiagnosed or misdiagnosed problem in soybean production fields. At Gainesville, severity of Fusarium wilt, or a complex which includes F. oxysporum, has reduced yields in some plots to near-ly zero

study of fatty acid synthase and adiponectin snps in the italian duroc breed

AbstractFatty acid synthase (FASN) is a multifunctional enzyme that plays a central role in fatty acid biosynthesis catalysing the conversion of acetyl-CoA and malonyl-CoA into long-chain saturated fatty acids and has an important role in energy homeostasis. Pig FASN gene has been assigned to chromosome 12p1.5 and a T>C polymorphism in the fourth exon was found. Adiponectin (ADN) is a fat-derived hormone involved in insulin sensitivity, in lipid and glucose metabolism. In literature is reported that the gene was mapped on chromosome 13 at 53.6 cM, in a region containing QTL for intramuscolar fat (IMF). In this gene several SNPs were identified and one of these polymorphisms (a G>A missense mutation within the 60th codon) determining the Val-Ile substitution in the protein, has been previously reported.The aim of this work is to analyse the variability of polymorphisms of fatty acid synthase described by Munoz et al., 2003 (Anim. Genet. 34:234) and adiponectin genes, candidates for meat and carcass quality..