Fine mapping of loci on BTA8 associated to antibody response to Mycobacterium avium paratuberculosis in cattle

Paratuberculosis (ParaTB) or Johne's disease, caused by Mycobacterium avium subspecies paratuberculosis commonly known as MAP in cattle, is a chronic gastroenteritis characterized by diarrhoea, decreased milk production and ultimately death. MAP is responsible for huge economic losses, particularly in dairy cattle herds. Susceptibility to MAP infection has been found to be heritable with heritability estimates ranging from 0.06 to 0.102. The definition of an infected animal can be based either on the presence of anti-MAP antibodies in the serum, or by direct demonstration of MAP in tissue or faeces by culture or PCR. Several studies have addressed the identification of genetic loci associated with MAP susceptibility. The objective of this study was to refine a locus associated with antibody response to Mycobacterium avium paratuberculosi (MAP). Using a genome- wide association analysis, a single nucleotide polymorphism on Bos taurus autosome BTA8 namely the SNP rs43161947 at posi- tion 35398490 with a p-value of 7.02 e-05, has previously been identified by the authors as associated with MAP infection. Fine mapping of the region was conducted with 100 single nucleotide polymorphisms spanning a region between BTA8: 34422912 and BTA8: 364553881 covering 2 Mega bases (Mb) designed in to cover 1 Mb ahead and after the SNP identified on BTA8. The 2 Mb region on BTA8 was evaluated within a group of 966 Holstein cows collected from routine ParaTB screening in the province of Lodi in Italy, in an area with a high prevalence of ParaTB. Animals were defined as ParaTB positive based on the detection of serum antibodies produced in response to MAP infection using the ID-screen\uae ELISA test (ID VET Montpellier, France). Of the 966 samples, 483 were MAP antibody positive (cases) and 483 MAP antibody negative (MAP negative controls). All animals were female, and cases and MAP negative controls were from the same farm tested on the same day.Using a single marker associ- ation analysis, conducted within the R statistical environment, we identified 3 different QTLs within the 2 Mega base region, under the main QTL on BTA8 associated with antibody response to MAP, in position 34.700.000, 35.800.000 and 36.400.000 bp. This reveals the complexity of the genetic architecture of thetrait and confirms the need to further explore the genome with fine mapping approaches, or by the use of whole genome sequencing to investigate complex traits, such as disease resistance

P316Expression and functional role of Ccdc80 in developing heart and in cardiomyopathies.

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TCO Optimization in Si Heterojunction Solar Cells on p-type Wafers with n-SiOx Emitter☆

Abstract Silicon heterojunction solar cells have largely demonstrated their suitability to reach high efficiencies. We have here focused on p-type c-Si wafers as absorber, considering that they share more than 90% of the solar cell market. To overcome some of the issues encountered in the conventional (n)a-Si:H/(p)c-Si configuration, we have implemented a mixed phase n-type silicon oxide (n-SiOx) emitter in order to gain from the wider bandgap and lower activation energy of this material with respect to (n)a-Si:H. The workfunction of the transparent conductive oxide layer (WTCO) plays also a key role, as it may induce an unfavourable band bending at the interface with the emitter. We have here focused on AZO, a promising alternative to ITO. Different layers with varying WTCO were prepared, by changing relevant deposition parameters, and were tested into solar cells. The experimental results have been explained with the aid of numerical simulations. Finally, for the n-SiOx/(p)c-Si heterojunction with optimized WTCO a potential conversion efficiency well over 23% has been estimated

The Utility of Capsicum annuum L. in Internal Medicine and In Dentistry: A Comprehensive Review

Capsaicin is a chili peppers extract, genus Capsicum, commonly used as a food spice. Since ancient times, Capsaicin has been used as a "homeopathic remedy" for treating a wild range of pathological conditions but without any scientific knowledge about its action. Several studies have demonstrated its potentiality in cardiovascular, nephrological, nutritional, and other medical fields. Capsaicin exerts its actions thanks to the bond with transient receptor potential vanilloid subtype 1 (TRPV1). TRPV1 is a nociceptive receptor, and its activation starts with a neurosensitive impulse, responsible for a burning pain sensation. However, constant local application of Capsaicin desensitized neuronal cells and leads to relief from neuropathic pain. In this review, we analyze the potential adjuvant role of Capsaicin in the treatment of different pathological conditions either in internal medicine or dentistry. Moreover, we present our experience in five patients affected by oro-facial pain consequent to post-traumatic trigeminal neuropathy, not responsive to any remedy, and successfully treated with topical application of Capsaicin. The topical application of Capsaicin is safe, effective, and quite tolerated by patients. For these reasons, in addition to the already-proven beneficial actions in the internal field, it represents a promising method for the treatment of neuropathic oral diseases

Insights in the genome of mycobacterium avium subsp. paratuberculosis by Next Generation Sequencing approaches

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis - or Johne's disease \u2013 that affects farmed and wild animals worldwide, causing negative economic consequences particularly relevant in the livestock sector of dairy cattle and beef (1). Recent estimates say that more than 50 % of the herds in Europe and North America are infected (3). In Italy, a study conducted in the Lombardy and Veneto regions reveals that about 70 % of dairy herds are infected (2). The disease shows high variability in the progression and symptoms that may be due to the genetic variability of the host, the pathogen, or a combination of the two. Understanding the mechanism responsible of this variability could be of paramount importance for the control of the disease (1, 3). Aim of this work was to study the genomic variability of MAP isolated from dairy cattle from different farms distributed in several Italian regions through the use of Next Generation Sequencing (NGS) techniques. The preliminary results on 15 strains are presented

s-ccdc80 is involved in axon guidance of zebrafish primary motoneurons

The similar to ccdc80 (s-ccdc80) gene was in silico identified on the basis of its high nuclear sequence identity (56,6%) with zebrafish ccdc80 (coiled-coil domain containing 80). The analysis of s-ccdc80 temporal/spatial expression by means of RT-PCR and whole mount in situ hybridization revealed its diffuse expression at early developmental stages and in epiboly. During somitogenesis up to 48 hours post fertilization, s-ccdc80 is expressed in specific territories such as cranial ganglia, ventral portion of somites, migrating adaxial cells and muscle pioneers, dorsal dermis and caudal vein plexus region. s-ccdc80 knock-down through the injection of a specific morpholino did not lead to morphological body plan defects nor alteration of the expression of several myogenic markers, though motility resulted affected. Thus, we decided to investigate the motoneuronal development in knocked-down embryos, also bearing in mind s-ccdc80 expression in territories involved in motoneurons patterning and axonal pathfinding: muscle pioneers and adaxial cells. s-ccdc80 knock-down led to axonal outgrowth defects of two primary motoneurons (CaP and Mip), that branched excessively and followed aberrant pathways. Our results strongly suggest that s-ccdc80 is involved in the guidance of the axonal extension of primary motoneurons but not in their proper formation. Furthermore, s-ccdc80 expression seemed to be positively regulated by the Hedgehog pathway

La risonanza magnetica nella programmazione chirurgica degli epatocarcinomi

The diagnostic accuracy of Magnetic Resonance Imaging (MRI) in the preoperative staging of unifocal hepatocellular carcinoma (HCC) was investigated and compared with that of ultrasonography (US) and Computed Tomography (CT). Eighteen patients with focal HCCs underwent MRI, CT and US scans before surgery. In all cases the histopathologic diagnosis was made with CT-guided fine-needle aspiration biopsy (FNAB). The diagnostic accuracy of each imaging modality was investigated with the assessment of three parameters thought to be of the utmost importance for surgical planning, i.e., lesion unifocality, the presence of a capsule and finally vascular involvement. MRI proved to be more sensitive than CT in demonstrating both lesion unifocality (100% vs. 94.4%) and the presence of a capsule (100% vs. 71.4%). In 2 of 18 patients some blood vessels were involved, which was clearly demonstrated only by MRI, CT missing it. Both MRI and CT had 100% specificity in the detection of a perilesional capsule and of vascular involvement. To conclude, MRI exhibited higher diagnostic accuracy than US and CT, thus confirming its major role in the preoperative staging of unifocal HCCs

Expression analysis and functional characterization of the dro1/cl2 gene during zebrafish somitogenesis

The recently cloned human DRO1/CL2 gene is expressed in almost all adult tissues and is downregulated in several cancer cell lines and solid tumors. DRO1/CL2 is indeed considered a potential oncosoppressor gene. To understand the physiological role of DRO1/CL2, we decided to clone its zebrafish homolog and to analyse its function during embryonic development. The dro1/cl2 zebrafish gene shares high homology with the mammalian counterpart and the translated protein (867 aa) presents the same structure, which is characterized by the simultaneous presence of a signal peptide and multiple nuclear localization signals as well as by three highly similar domains of still unknown function. dro1/cl2 is maternally supplied. Temporal expression analysis performed by RT-PCR showed that it is downregulated during gastrulation. Its expression level starts increasing from the initial steps of segmentation up to the late somitogenesis, when it reaches the highest value. Whole mount in situ hybridization experiments show a diffuse dro1/cl2 expression during cleavage and epiboly. Interestingly, during somitogenesis, the dro1/ cl2 expression becomes mainly restricted to the notochord. Finally, in later developmental stages it becomes detectable in the muscles of both trunk and head, and in the heart. To investigate the functional role of the dro1/cl2 gene, we performed loss- and gain-of-function experiments by injecting a specific dro1/cl2 MO and the full length mRNA, respectively. The MO injected embryos presented with abnormally shaped somites in the early somitogenesis, while curved tails were evident at the end of this developmental phase. dro1/cl2 overexpression also impaired somitogenesis. To determine whether the observed defects are a result of a direct activity of dro1/cl2 on the different phases of somitogenesis rather than an indirect consequence of a disruption of nothocordal signals, we evaluated the expression of notochordal and somite markers in the injected embryos. We excluded a dro1/cl2 functional role in the developing notochord. Indeed, no tail and sonic hedgehog were unaffected. By converse, the block of the Hh signals indicated dro1/cl2 as a target of the Hh pathway. The expression pattern of the myogenic markers myod and myogenin resulted heavily disrupted in the lateral domain without affecting the adaxial one. The segmentation clock marker her1, and the segment polarization markers mesp-a and mesp-b showed no significant alteration. Interestingly, while no apparent effect was seen on the expression pattern of gadd45beta2 in the furrow, we observed the disappearance of its expression in the already formed somites. The demonstration of a specific link between dro1/cl2 and gadd45beta2 expression derived from rescue experiments, showing a significant phenotype recovery in dro1/cl2 morphants injected with gadd45beta2 mRNA. In our knowledge, this is the first demonstration of a notochordal signal controlling the lateral somitic maturation through gadd45beta2

The Coiled-Coil Domain Containing 80 (ccdc80) Gene Regulates gadd45β2 Expression in the Developing Somites of Zebrafish as a New Player of the Hedgehog Pathway

The Coiled-Coil Domain Containing 80 (CCDC80) gene has been identified as strongly induced in rat thyroid PC CL3 cells immortalized by the adenoviral E1A gene. In human, CCDC80 is a potential oncosoppressor due to its down-regulation in several tumor cell lines and tissues and it is expressed in almost all tissues. CCDC80 has homologous in mouse, chicken and zebrafish. We cloned the zebrafish ccdc80 and analyzed its expression and function during embryonic development. The in silico translated zebrafish protein shares high similarity with its mammalian homologous, with nuclear localization signals and a signal peptide. Gene expression analysis demonstrates that zebrafish ccdc80 is maternally and zygotically expressed throughout the development. In particular, ccdc80 is strongly expressed in the notochord and it is under the regulation of the Hedgehog pathway. In this work we investigated the functional effects of ccdc80-loss-of-function during embryonic development and verified its interaction with gadd45\u3b22 in somitogenesis