http://http://dspace.unitus.it:80 The DSpace digital repository system captures, stores, indexes, preserves, and distributes digital research material. 2013-05-22T20:35:44Z http://hdl.handle.net/2067/1842 Title: Proteomics and Transcriptomics Investigation on longissimus Muscles in Large White and Casertana Pig Breeds Authors: Murgiano, Leonardo; D'Alessandro, Angelo; Egidi, Maria Giulia; Crisa, Alessandra; Prosperini, Gianluca; Timperio, Anna Maria; Zolla, Lello Abstract: 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. Description: L'articolo è disponibile sul sito dell'editore http://pubs.acs.org/ 2009-12-31T23:00:00Z http://hdl.handle.net/2067/1848 Title: Comparative proteomics and transcriptomics analyses of livers from two different Bos taurus breeds: “Chianina and Holstein Friesian” Authors: Timperio, Anna Maria; D'Alessandro, Angelo; Pariset, Lorraine; D'Amici, Gian Maria; Valentini, Alessio; Zolla, Lello Abstract: The Holstein Friesian and Chianina cattle breeds are representative of extreme selection for milk and meat traits, respectively, with significant changes in metabolism resulting from human selection over the past centuries. In the present study, we wanted to assess whether selection for different purposes has had a measurable effect on liver metabolism through a comparison of the protein and gene expression profiles of the two breeds. We applied 2-DE in order to identify proteins which were differentially expressed in the livers of the two breeds and relate them to different liver functions. We expected to find that only a small number of proteins would be differentially expressed, due to the relatively short phylogenetic distance between these cattle breeds. Nonetheless, thirty nine differentially-expressed proteins were characterized between Chianina and Holstein Friesian, out of a total of 560 ± 57 spots that matched. Microarray analyses evidenced the differential expression of 167 genes (148 for the Holstein Friesian and 19 for the Chianina). Despite being closely related at the genetic level, the disparity of the proteomic and transcriptomic profiles of these two breeds allows us to perform pathway analysis thus to pinpoint proteins whose expression might render the latter capable of greater milk production, or proteins involved in altered thermoregulatory ability or hormone production. On the other hand, we found proteins and gene transcripts in Chianina, not expressed in Holstein, which, upon interaction pathway analysis, were mainly involved in anabolic pathways. In brief, our integrated study provides molecular evidences to support the physiological differences between Holstein and Chianina cattle breeds. Description: L'articolo è disponibile sul sito dell'editore http://www.sciencedirect.com/ 2008-12-31T23:00:00Z http://hdl.handle.net/2067/1843 Title: Recombinant clotting factor VIII concentrates: Heterogeneity and high-purity evaluation Authors: D'Amici, Gian Maria; Timperio, Anna Maria; Gevi, Federica; Grazzini, Giuliano; Zolla, Lello Abstract: Factor VIII is an important glycoprotein involved in hemostasis. Insertion of expression vectors containing either the full-length cDNA sequence of human factor VIII (FLrFVIII) or B-domain deleted (BDDrFVIII) into mammalian cell lines results in the production of recombinant factor VIII (rFVIII) for therapeutic usage. Three commercially available rFVIII concentrates (Advates, Helixate NexGens and Refactos), either FLrFVIII or BDDrFVIII, were investigated by 1- and 2-DE and MS. The objective of this study was to compare the heterogeneity and the high purity of both rFVIII preparations before and after thrombin digestion. In particular, the 2-D gel was optimized to better highlight the presence of contaminants and many unexpected proteins. Recombinant strategies consisting of insertion of expression vectors containing BDDrFVIII and FLrFVIII resulted in homogeneous and heterogeneous protein products, respectively, the latter consisting in a heterogeneous mixture of various B-domain-truncated forms of the molecule. Thrombin digestion of all the three rFVIII gave similar final products, plus one unexpected fragment of A2 domain missing 11 amino acids. Regarding the contaminants, Helixate NexGens showed the presence of impurities, such as Hsp70 kDa, haptoglobin and proapolipoprotein; Refactos showed glutathione S-transferase and b-lactamase, whereas Advates apparently did not contain any contaminants. The proteomic approach will contribute to improving the quality assurance and manufacturing processes of rFVIII concentrates. In this view, the 2-DE is mandatory for revealing the presence of contaminants. Description: L'artcoo è disponibile sul sito dell'editore http://onlinelibrary.wiley.com/ 2009-12-31T23:00:00Z http://hdl.handle.net/2067/1844 Title: Comparison among plasma-derived clotting Factor VIII by using Authors: Timperio, Anna Maria; Gevi, Federica; Grazzini, Giuliano; Vaglio, Stefania; Zolla, Lello Abstract: Background. Deficiency or dysfunction of coagulation factor VIII (FVIII) is the underlying cause of haemophilia A. Haemophilic patients are at present treated with plasma-derived FVIII (pdFVIII) or recombinant FVIII (rFVIII) in order to correct their clotting deficiency. pdFVIII concentrates are exclusively produced from human plasma upon pooling from multiple donors. It is not know whether the presence of excess of other plasma proteins, in addition to von Willebrand factor, could stimulate untoward immune responses in the recipient. Thus, information regarding the presence of contaminants in commercial products is of concern. Materials and Methods. Two commercially available pdFVIII concentrates were characterized through SDS-PAGE and mass spectrometry Emoclot® and Beriate® . Results. The components of two pdFVIII products considered in this study were well identified by mass spectrometry analysis, in both cases we found abundant components coming from blood plasma, and some other contaminants. Only in Beriate® we also found truncated form of pdFVIII. Conclusion. The two pdFVIII examined showed the presence of vWF, Fibrinogen in excess, and other substances that could be considered as contaminants or impurities. Description: L'articolo è disponibile sul sito dell'editore http://www.bloodtransfusion.it 2009-12-31T23:00:00Z http://hdl.handle.net/2067/1887 Title: Comparative proteomics and transcriptomics analyses of livers from two different Bos taurus breeds: “Chianina and Holstein Friesian” Authors: Timperio, Anna Maria; D'Alessandro, Angelo; Pariset, Lorraine; D'Amici, Gian Maria; Valentini, Alessio; Zolla, Lello Abstract: The Holstein Friesian and Chianina cattle breeds are representative of extreme selection for milk and meat traits, respectively, with significant changes in metabolism resulting from human selection over the past centuries. In the present study, we wanted to assess whethe selection for different purposes has had a measurable effect on liver metabolism through a comparison of the protein and gene expression profiles of the two breeds. We applied 2-DE in order to identify proteins which were differentially expressed in the livers of the two breeds and relate them to different liver functions. We expected to find that only a small number of proteins would be differentially expressed, due to the relatively short phylogenetic distance between these cattle breeds. Nonetheless, thirty nine differentially expressed proteins were characterized between Chianina and Holstein Friesian, out of a total of 560 ± 57 spots that matched. Microarray analyses evidenced the differential expression of 167 genes (148 for the Holstein Friesian and 19 for the Chianina). Despite being closely related at the genetic level, the disparity of the proteomic and transcriptomic profiles of these two breeds allows us to perform pathway analysis thus to pinpoint proteins whose expression might render the latter capable of greater milk production, or proteins involved in altered thermoregulatory ability or hormone production. On the other hand, we found proteins and gene transcripts in Chianina, not expressed in Holstein, which, upon interaction pathway analysis, were mainly involved in anabolic pathways. In brief, our integrated study provides molecular evidences to support the physiological differences between Holstein and Chianina cattle breeds. 2011-04-05T22:00:00Z http://hdl.handle.net/2067/1880 Title: Comparison of Milk Fat Globule Membrane (MFGM) Proteins of Chianina and Holstein Cattle Breed Milk Samples Through Proteomics Methods Authors: Murgiano, Leonardo; Timperio, Anna Maria; Zolla, Lello; Bongiorni, Silvia; Valentini, Alessio; Pariset, Lorraine Abstract: Identification of proteins involved in milk production is important to understand the biology of lactation. Many studies have advanced the understanding of mammary function and milk secretion, but the critical molecular mechanisms implicated in milk fat secretion is still incomplete. Milk Fat Globules are secreted from the apical surface of the mammary cells, surrounded by a thin membrane bilayer, the Milk Fat Globule Membrane (MFGM), formed by proteins which have been suggested to be holesterolemia-lowering factors, inhibitors of cancer cell growth, vitamin binders, bactericidal, suppressors of multiple sclerosis. Using a proteomic approach, we compared MFGM from milk samples of individuals belonging to two different cattle breeds, Chianina and Holstein,representative of selection for milk and meat traits, respectively. We were able to isolate some of the major MFGM proteins in the examined samples and to identify differences between the protein fractions of the two breeds. We detected differences in the amount of proteins linked to mammary gland development and lipid droplets formation, as well as host defence mechanisms. We have shown that proteomics is a suitable, unbiased method for the study of milk fractions proteins and a powerful tool in nutritional genomics. 2011-04-05T22:00:00Z http://hdl.handle.net/2067/1847 Title: Preface Authors: Zolla, Lello Abstract: Since in the last few years clinicians and proteomicists have rapidly reached the rendez-vous point, we are currently in the middle of the docking manoeuvres. Two formerlyindependent worlds have started to cooperate, with the declared intent to ameliorate the quality of the production and handling processes of blood components and plasma derivatives, with the shared goal to guarantee their safety and effectiveness for the whole health care system. This special issue tries to add some little pieces to this yet complicated puzzle, since it represents an attempt to sail across the mare magnum of the actual knowledge in this research endeavour, to the end of summarizing the main advancements. In the light of what emerges, blood-related proteomics appears as much as promising as never before. In particular, proteomics has been recently transformed from a mere basic-research expensive toy into a dramatically-sensitive and high-throughput research tool which could be exploited to investigate the molecular mechanisms of blood and blood components and promises to establish quality parameters in the blood-banking production-chain totally anew. Description: L'articolo è disponibile sul sito dell'editore http://www.sciencedirect.com/ 2009-12-31T23:00:00Z