The purposes of this research are to find out consumer¡¦s perception and expectation of beef safety. Our questionnaire research is by asking people who lives in Kaohsiung city and had bough beef before to find out those factors which affect purchasing willing. Our research contents two purposes. The first is to understand the consumer¡¦s perception of import beef safety. The other is to find out the factors those affect buying willing. The total responses are 204 questionnaires. Use ANCOVA and regression statistics to analyze our data. Results showed that: (1) age is significant to the knowledge of BSE. (2) the belief of government and Prof. who has positive attitude about BSE is significant to purchasing willing. (3) percentage of relatives and friends who also eat beef is significant to purchasing willing. According to our research results, we made some suggestions to government to improve beef permit system and to promote knowledge of BSE to consumer.

Investigations detailed in this thesis constitue a part of continuing programme of research work undertaken in this laboratory on vitamin binding proteins. Avidin from the chicken egg white, streptavidin &om the bacterium Streptromyces avidin and biotin binding proteins (BBP-I and BBP-11) from chicken egg yolk constitute a family of proteins that bind the vitamin biotin with extremely high affinities. The yolk BBPs are involved in the deposition of the vitamin in the developing oocyte in chicks whereas an antimicrobial function has been attributkl to avidin.. The fact that all these proteins bind the vitamin in the same manner, unlike biotin-dependent enzymes, indicates that the structural features involved in ligand binding could be similar, if not identical in these proteins. To delineate the basis of putative structural similarity among these proteins, studies were carried out using antibodies as the immunological probes. Avidin, a homotetremer glycoprotein, with a subunit Mr of 17,000 has been purified to homogeneity from chicken egg white using a novel procedure involving ammonium sulphate fractionation, ethanol precipitation and S-Sepharose column chromatography. Despite their lesser abundance in chicken egg yolk associated with a large amount of interfering lipids during the purification, both BBP-I (monomer and shown to be precursor for BBP-11) and BBP-I1 (tetramer) have been purified to homogeneity by employing a common method using butanol extraction to remove the lipids, DEAE-Sephacel column chromatography, biotin-AH-Sepharose affinity chromatography and fast performance liquid chrometography (FPLC) system. The purity of all these proteins was confirmed by SDS-PAGE analysis.

Texte remanié de: Thèse de doctorat--Histoire--Limoges, 2007. Titre de soutenance : Une spécialisation bovine en pays de petite culture : l'élevage bovin en Limousin au XVIIIe siècle. / En appendice, choix de documents. Bibliogr. p. 407-437. Notes bibliogr. Index.

Thèse d'exercice : Pharmacie : Université de Nantes : 2004. / Bibliogr. f. 110-117 [163 réf.].

Thèse d'exercice : Pharmacie : Université de Nantes : 2004. / Bibliogr. f. 84-88 [39 réf.].

Rapport de recherche bibliographique (DPSSIB) : Ecole nationale supérieure des sciences de l'information et des bibliothèques : Villeurbanne (France) : 2001. / DPSSIB = Diplôme professionnel supérieur en sciences de l'information et des bibliothèques. Notes bibliogr.

Bovine adenovirus (BAdV)-3, a Mastadenovirus was isolated from the healthy and sick cattle (Darbyshire et al., 1965; Zhu et al., 2011). Like other adenoviruses, BAdV-3 replication is characterized by the temporally regulated expression of genes characterized by early, intermediate and late gene expression. Genus-common, non-structural protein 100K is encoded by late region L6 of BAdV-3. The objective of the present study was to characterize the BAdV-3 100K protein and identify cellular and viral proteins interacting with 100K. Although BAdV-3 100K encoded as 850 amino acid polypeptide (Reddy et al., 1998), rabbit antisera raised against peptides representing N-terminus or C-terminus recognized a protein of 130 kDa at 12-24 hrs post infection, and proteins of 130 kDa, 100 kDa, 95 kDa and 15 kDa at 36-48 hrs post infection. The 100K appeared to be localized to the nucleus and cytoplasm of BAdV-3 infected cells. In contrast, 100K localized predominantly to cytoplasm of transfected cells. However, BAdV-3 infection of cells transfected with 100K-EYFP expressing plasmid detected fluorescent protein in nucleus of the cells suggesting that another viral protein may be required for the nuclear localization of 100K. Using yeast two-hybrid and GST pull-down assays, 100K protein was shown to interact with BAdV-3 33K protein. These results were validated using bimolecular fluorescence complementation (BiFC) assay. Although, 100K protein interacts with 33K protein, co-expression of both proteins in transfected cells did not alter the cytoplasmic localization of 100K. Using GST-pull down assay and BiFC assay, 33K interacting region of 100K was localized to a stretch of 13 amino acids (624-637). Repeated attempts were not successful in rescuing a recombinant BAdV-3 expressing mutant 100K (containing deletion of amino acids 624-637). The interaction of cellular protein(s) with 100K was determined by mass spectrometric analysis of immunoprecipitated 100K. Mass spectrometry of immunoprecipitate obtained by immunoprecipitating 100K protein from BAdV-3 infected cells harvested at 48 hrs post infection identified six proteins including dynein light chain (DYNLT)1. The initial identified interaction of 100K with DYNLT1 was confirmed by the yeast two-hybrid assay, co-immunoprecipitation assay and BiFC assays. Furthermore, DYNLT1 interacting domain of 100K protein of BAdV-3 was found to be located between 499-587 amino acids. Co-expression of BAdV-3 100K-EY fusion protein with myc epitope tagged DYNLT1 protein did not alter the localization of 100K-EY fusion protein. The investigation into the differences in the subcellular localization of the 100K protein in the transfected and infected cells lead to identification of the cleavage by adenoviral protease. Subsequent analysis suggested that BAdV-3 protease cleaves 100K at two identified potential protease cleavage sites (amino acid 740-745 and 781-786) in transfected or BAdV-3 infected cells. Although protease encoded by human adenovirus (HAdV)-5 or porcine adenovirus (PAdV)-3 also cleaved BAdV-3 100K at potential identified protease cleavage sites, no such cleavage of 100K encoded by HAdV-5 or PAdV-3 could be detected in cells expressing virus specific protease. Successful isolation of recombinant BAdV-3 expressing mutant protease (substitution of alanine for glycine in potential protease cleavage site) suggested that cleavage of BAdV-3 100K by viral protease is not essential for viral replication. However, further analysis observed less virus in the supernatant of cells infected with mutant BAdV-3 compared to WT BAdV-3 suggesting a possible role for cleaved C-terminal fagment in lysis of infected cells. Co-expression of BAdV-100K with other late viral proteins suggested that the 100K-EYFP fusion protein localized to the nucleus in cells co-expressing BAdV-3 protease-DsRed fusion protein. Interestingly, only C-terminal cleaved fragment of 100K localizes to the nucleus in BAdV-3 protease expressing cells. Further analysis suggested that C-terminal fragment localizing to the nucleus contains a bipartite nuclear localization signal, which is recognized by importin α3. Our results suggest that the N-terminal part of 100K may be retained in the cytoplasm by interaction with Tctex1 (DYNLT1). Our study provides for the first time a plasmid co-transfection system for the study of the protease cleavage of viral proteins. Moreover, this is the first report of cleavage of any non-structural viral proteins by adenoviral protease in infected cells.

The goal of this dissertation was to evaluate genomic regulation during bovine mammary epithelial cell (BMEC) growth and differentiation. To accomplish this goal, a collagen gel cell culture system was developed that was capable of mimicking the prepartum stages of epithelial development and differentiation. In addition, a 4,600-cDNA bovine microarray was developed in order to profile gene expression. Analysis of BMEC in collagen cultures using various lactogenic conditions highlighted the critical importance of both hormonal and structural signals. The objective of the first study utilizing the microarray was to evaluate the contribution of the two prominent lactogenic factors in vitro, 1) prolactin and 2) gel release. Collectively, lactogenic stimulation appears to turn off genes associated with structural progression and morphogenesis, and turn on genes involved in alveolar MEC differentiation such as cell polarization, milk protein synthesis and ER/Golgi transport. The objective of the second study utilizing these resources was to evaluate the direct effects of thermal stress on BMEC growth and development. The structural response to thermal stress was characterized by morphogenic inhibition and dramatic regression of the ductal branches. Microarray analysis revealed an overall up-regulation of genes associated with stress response, DNA repair, protein degradation and cell death. In contrast, genes associated with cellular and MEC-specific biosynthesis, metabolism, and morphogenesis, were generally down-regulated. Subsequent to the analysis of BMEC differentiation was a targeted effort focusing on two small molecules hypothesized to be involved in regulating the BMEC secretory response: serotonin and prostaglandin E2. A pilot study suggested that serotonin is produced by bovine MEC and a model was proposed that describes serotonin's role as a feedback inhibitor during milk synthesis and secretion. A second pilot study demonstrated that PGE2 had a consistently positive influence on lumen diameter of alveolar structures in vitro. Overall, this dissertation provides new resources for studying bovine functional genomics, particularly within the mammary gland, and it provides a strong foundation for understanding genomic regulation of mammary epithelial structure and function. Furthermore, it establishes potential roles for local regulation of milk production by serotonin and PGE2.

To understand bovine oocytes meiotic maturation, developmental potential, gene expression is required. The gene expression studies in the preimplantation bovine oocytes has been difficult, because the procedures that are being employed for extracting total RNA are not specific for bovine oocytes and so far is not providing the required amount for further procedures. Quantification of genes generally requires large amounts of total RNA in order to overcome the problem of low amount of mRNA present, so a standardized specific protocol is recommended. These days most of the researchers are using commercial Kit protocols without knowing the significance of chemicals and how they are acting on cells. In present project a standardized protocol (modified trizol) was designed for bovine oocytes, which was specific and less expensive. The efficiency of this protocol compared with Pure Link (Kit Protocol), GNTC (Guanidinium thiocyanate) for extraction of total RNA from fresh oocytes, vitrified oocytes with PROH (1,2 propanediol) and DMSO (dimethylsulfoxide) cryoprotectans was much better. The RNA (absorbance 260/280) purity levels of the standardized protocol was ranging (1.50-2.10), whereas for GNTC protocol (1.05-1.36), Pure Link (kit protocol) (2.05-2.7). Amplification of housekeeping genes (SDHA and GAPDH gene) showed the specificity and efficiency of the standardized protocol over other protocols.

Assessment of occurrence and diagnostic methods of viral diseases in cattle – viral diarrhoea (BVD), infectious bovine rhinotracheitis (IBR), rabies, enzootic bovine leukosis (EBL), and spongiform encephalopathy (BSE) – was carried out for the first time in Lithuania. It was established that viruses of rabies, infectious rhinotraheitis and viral diarrhoea are most widespread in the country. It was determined that occurrence of rabies in cattle is parallel with the infection of wildlife with rabies virus. Analysis of eradication programme of enzootic bovine leucosis was done. It revealed that only combined application of diagnostic and preventive measures allowed reducing the cattle infection up to 0.2%. Though bovine spongiform encephalopathy has not been recorded in Lithuania, it is feasible to implement its diagnostic and prevention programme. An overall financial analysis of expenditures on BSE and viral diseases diagnostics and control was for the first time done in Lithuania. It showed that BSE and EBL occupied the leading positions in the structure of expenditures on viral diseases. In 2001, expenditures on BSE investigations accounted for 76.68% and in 2004 for 86.74% of the total. Expenditures on EBL investigations relatively reduced from 86.98% in 2000 to 8.47% in 2004. During the time under consideration, expenditures on investigations of other viral diseases changed but little. It was determined that consistent and wide-scale preventive vaccination created... [to full text]