Strategy to isolate, in vivo, a eukaryotic replication origin for the analysis of its asscoiated proteins

In all organisms the DNA content is precisely and efficiently replicated. DNA replication starts at specific regions called replication origins, where a preReplicative Complex (pre-RC) is assembled at the end of the G2/M phase, maldng chromatin competent for replication. At the transition between G1 and S phases the activation of two conserved protein kinases triggers the transition from pre-RC into the active form, the Replicative Complex (RC). Replication origins are not fired at the same time during S phase, thus we can define origins as “early” and “late” according to their time of activation. During the past years our knowledge about DNA replication has improved due to the identification of novel proteins. Despite a large body of work, it remains possible that additional factors remain unidentified. The discovery of new proteins by genetic means is difficult due to the presence of redundant mechanisms and to their requirement for cell survival. Thus the aim of my project is to develop an unbiased assay to study, in Saccharomyces cerevisiae, the proteome of a single eukaryotic replication origin and identify novel proteins involved in DNA replication. Because the chromosomal environment profoundly influences the behaviour of replication origins (such as timing, efficiency), it is therefore of interest to analyse the proteomics of replication origins in their endogenous context. Taking advantage of the pSRl recombination system, I constructed two plasmids (pLTl/RsiteA and pLT2/RsiteB) that led to the integration of recombination sites into the yeast genome and the excision in vivo of the late replication origin ARS1413. These plasmids, allowing the excision of any flanking region, represent important genetic tools. The resulting episome has been isolated from the genome by density gradient purification in order to analyze by mass spectrometry the protein associated with the specific replication origin. This technique can be used to study the proteins involved in different steps of DNA replication. In fact cells arrested in G1 with a-factor will provide the proteome of the pre-RC while cells treated with HU will represent the proteome of late origins that are inhibited by the S-phase checkpoint

Racing for radical innovation:how motorsport companies harness network diversity for discontinuous innovation

The motorsport industry is a significant part of the UK economy. According to industry estimates approximately 4,500 companies are involved in the UK Motorsport and Performance Engineering Industry and its wide-ranging support activities. The industry has an annual turnover of £6.0 billion, and contributes £3.6 billion worth of exports. The Motorsport Industry Association estimates that the support side of the sector alone "involving events management, public relations, marketing, sponsorship and a host of other support functions" accounts for approximately £1.7 billion of the yearly industry total. And in terms of employment, UK Motorsport supports 38,500 full and part-time jobs, including 25,000 engineers

Flesh qualitative traits of tub gurnard (Chelidonichthys lucerna L.), a promising species candidate for aquaculture, captured in the middle Adriatic Sea in different seasons

The quality traits of the fillets from tub gurnard (Chelidonichthys lucerna L.) fished in the mid-Adriatic Sea were investigated. Forty fishes per season were sampled to evaluate the proximate composition, cholesterol content and fatty acid profile of fillets. Seasons significantly affected the quality traits of flesh. The protein content ranged from 19.39% in winter to 19.67% in summer, without significant differences. Lipid content was notably higher in spring (2.28%) and summer (2.32%), compared to autumn (1.72%) and winter (1.31%). Energy content was significantly higher in spring (416.45 kJ/100 g) and summer (417.97 kJ/100 g) compared to autumn (391.35 kJ/100 g) and winter (372.79 kJ/100 g). Saturated fatty acid content was highest in spring (35.88%), whereas monounsaturated acid content was not influenced by season. The n-3 polyunsaturated fatty acid content exceeded 37% of total fatty acid content during summer. The n-6/n-3 ratio remained favourably low across all seasons (from 0.16 to 0.18), with a slight significant increase in autumn (0.31). In conclusion, this study indicates that the flesh of tub gurnard has high nutritional value year-round, with the best results (in terms of n-3 polyunsaturated fatty acids) being obtained in summer

Idiopathic Eosinophilic Pneumonia with Associated Pulmonary Vasculitis in Horse: A Case Series

Various eosinophilic lesions and their association with diseases have been described in horses. Multisystemic Eosinophilic Epitheliotropic Disease (MEED) characterized by eosinophilic granulomas in various organs represent the most diffuse manifestation. In the present study, we describe the gross and microscopic lesions of idiopathic eosinophilic pneumonia with associated pulmonary vasculitis in 11 horses without systemic involvement. Lungs were enlarged, pale pink, poorly collapsed with white-reddish nodules, randomly distributed throughout the parenchyma. Histologically the lesions ranged from mild eosinophilic bronchointerstitial pneumonia to severe eosinophilic lobular bronchopneumonia associated with eosinophilic necrotizing vasculitis affecting small to medium-size vessels. There was no histologic evidence of parasites within the lesion of any of these horses. Eosinophilic pneumonia with vasculitis and without intralesional parasites have not been previously reported in horses. The current histologic findings are similar to pathologic lung findings observed in Churg-Strauss syndrome of man

Indagini preliminari sull’espressione immunoistochimica della proteina p53 nel seminoma del cane

SUMMARY - Tlie p53 protein is a transcription factor inducing celi cycle arresi and apoptosis against a DNA damage. p53 mutations might lead to lose its inibitoring growth functions and contribute to not controlied cellular proliferation. Mutations and/or overexpression of p53 are thè most comnion genetic alterations found in human cancer. The role of thè p53 protein in cancerogenesis m domestic animals is stili largely unknown. This prelimmary study investigates thè expression of p?3 in 16 semmomas of thè dog using a standard streptavidinbiotin peroxidase method after microwave antigen retrieval. In 9 diffuse seminomas tlie mtensity of p53 expression is of low degree (^), in 5 diffuse seminomas is of middle degree (++) and m 2 intratubular seminomas is of higt degree (+++). The results suggest tlie p53 expression m seminoma of thè dog decreases with advancing stage of neoplastic process

Suitability of artisanal fishery discards as feed for juvenile tub gurnard (chelidonichthys lucerna L.) reared in sea bottom cages in the mid Adriatic Sea

The suitability of using discards from artisanal fisheries as feed for wild-caught Chelidonichthys lucerna (L.) reared in submerged cages in the Adriatic Sea was investigated. Three-hundred juvenile tub gurnard (72.9 ± 11 g, 18 ± 1 cm) were captured and separated into four circular 35.3 m3 submerged cages. Two stocking densities were used: two cages contained 60 fish each (1.7 fish m-3; CG-L) and two cages contained 90 fish each (2.55 fish m-3; CG-H). Growth performance and survival rates were recorded over a 240 day period during which tub gurnard were fed with a variety non-target fishery discards, including common crab, sardine, Atlantic chub mackerel, anchovy, and salema. The survival rate was approximately 90% in both groups (CG-L = 91% and CG-H = 90%), with final stocking densities of 0.32 kg m-3 and 0.49 kg m-3 for the CG-L and CG-H groups, respectively. There were no significant differences in final mean body weight or length between the CG-L group (206 ± 23 g, 24.6 ± 2 cm) and the CG-H group (215.5 ± 32 g, 24.8 ± 2 cm). The condition index was similar between the two groups (1.38 and 1.42 for CG-L and CG-H, respectively). The feed conversion ratio was high in both the CG-H (2.39) and CG-L (2.32) cages. These results demonstrate that viable growth rates of tub gurnard may be obtained by feeding recycled fishery discards and rearing in cages placed on the seabed, which allows for the natural benthic behavior of this species

Origin of Enzootic Intranasal Tumor in the Goat (Capra hircus): A Glycohistochemical Approach

Enzootic intranasal tumor (EIT) appears glandular in type and has recently been classified as an adenocarcinoma of low malignancy. The aim of this study was to characterize the secretion of surface glycoconjugates (GCs) in EIT and in normal respiratory and olfactory mucosae of the goat by means of conventional and lectin histochemistry, in order to shed light on the histogenesis of EIT. Morphologic and ultrastructural investigations showed two growth types of EIT: i.e., tubular and papillary patterns. Conventional histochemistry revealed the presence of neutral and carboxylated GCs in the olfactory glands and in the tubular part of EIT, as well neutral and sulphated GCs in the respiratory glands and in the papillary part of EIT, suggesting that the papillary pattern tumor arises from the respiratory glands, whereas the tubular portion of EIT arises from the olfactory glands. Lectin histochemistry gave further information on the expressed GCs

Immunohistochemical Expression of p62 in Feline Mammary Carcinoma and Non-Neoplastic Mammary Tissue

The p62 protein, also called sequestosome 1 (SQSTM1), is a ubiquitin-binding scaffold protein. In human oncology, although the interest in the function of this protein is recent, the knowledge is now numerous, but its role in tumorigenesis is not yet clear. This preliminary study aims to evaluate the immunohistochemical expression of p62 in 38 cases of feline mammary carcinoma with different grades of differentiation and in 12 non-neoplastic mammary gland tissues, to assess the expression level and a possible correlation with malignancy. The expression of p62 was statistically higher in carcinoma compared to non-neoplastic mammary glands: 28 feline mammary carcinomas (73.7%) had a high p62 expression score, three (7.9%) had a moderate expression, while seven cases (18.4%) had a low expression. The grade of the differentiation of the carcinoma was not correlated with the p62 expression. This study represents the first approach in feline oncology that correlates p62 expression in feline mammary carcinoma. Our results, although preliminary, are similar to the results of human breast cancer, therefore, also in the cat, p62 could be considered a possible oncotarget

Preliminary survey of the nutritional and palynological traits of honey bee-foraged pollen from Liguria (Italy)

It is known that the palynological features, and consequently the nutritional quality, of bee-foraged pollen vary according to several factors such as sampling site and period of year. In our two year study, carried out as part of the COLOSS CSI Pollen study between April and September in the eastern province of Genova (Italy), we explored the relationships between the palynological spectra and the protein content of corbicular pollen. We found that the differences are mainly driven by the collection season rather than by the site. This observation allowed us to employ the sample\u2019s palynological traits for the identification of foraging seasons characterized by different protein content in pollen. The outcome of our study provides a useful tool for beekeepers, helping them to cautiously program certain apiary management practices. This work also provides suggestions for future monitoring programs, regarding the advantages of studying the seasonal pattern not only of the floral origin of pollen but also of its nutritional traits