Structural Basis for Chaperone-Independent Ubiquitination of Tau Protein by Its E3 Ligase CHIP.

The multi-site ubiquitination of Tau protein found in Alzheimer's disease filaments hints at the failed attempt of neurons to remove early toxic species. The ubiquitin-dependent degradation of Tau is regulated in vivo by the E3 ligase CHIP, a quality controller of the cell proteome dedicated to target misfolded proteins for degradation. In our study, by using site-resolved NMR, biochemical and computational methods, we elucidate the structural determinants underlying the molecular recognition between the ligase and its intrinsically disordered substrate. We reveal a multi-domain dynamic interaction that explains how CHIP can direct ubiquitination of Tau at multiple sites even in the absence of chaperones, including its typical partner Hsp70/Hsc70. Our findings thus provide mechanistic insight into the chaperone-independent engagement of a disordered protein by its E3 ligase.

The Impact of "Wine Country of Origin" on the Perception of Wines by South African and French Wine Consumers: A Cross-Cultural Comparison.

Culture is an important factor that influences how marketing interacts with food choice. This study aims at exploring the effect of consumers' Country of Origin (COO) on wine representations and perception using Chenin blanc as a model. The first objective was to evaluate the role of origin in the construction of the representation. We used the theoretical framework of social representation to compare South African (SA) and French consumers' representations via a word association task. The results indicated that SA representations are dominated by consumers' experience of the wine (sensory and emotional dimensions), whereas French representations are dominated by the wine itself, in particular its origin and mode of consumption. The second objective was to evaluate the effect of origin on wine categorization in two conditions: with and without information concerning the two geographical origins of the samples. Results showed that providing information on the origin of the wines affected French participants more than SA participants. In both conditions, the groups of wines formed in the sorting tasks by SA participants were based on sensory descriptors and appeared not to be impacted by the information on origin. In contrast, providing information on the origin of the wines to French participants led to an increased use of the words "Loire", "South Africa" and "familiar" suggesting a different sorting strategy more deliberately based on the origin of the wines. Our findings have important implications for the marketing and export activities within the wine industry.

Modelling the sensory space of varietal wines: Mining of large, unstructured text data and visualisation of style patterns.

The increasingly large volumes of publicly available sensory descriptions of wine raises the question whether this source of data can be mined to extract meaningful domain-specific information about the sensory properties of wine. We introduce a novel application of formal concept lattices, in combination with traditional statistical tests, to visualise the sensory attributes of a big data set of some 7,000 Chenin blanc and Sauvignon blanc wines. Complexity was identified as an important driver of style in hereto uncharacterised Chenin blanc, and the sensory cues for specific styles were identified. This is the first study to apply these methods for the purpose of identifying styles within varietal wines. More generally, our interactive data visualisation and mining driven approach opens up new investigations towards better understanding of the complex field of sensory science.

[Application of pulsed-field gel electrophoresis (PFGE) to methicillin-resistant strains of Staphylococcus aureus from humans and domestic animals]. / Applicazione della elettroforesi pulsata (PFGE) a stipiti meticillino-resistenti di Staphylococcus aureus isolati dall'uomo e dagli animali.

BACKGROUND: The aim of the research was to isolate and to identify the methicillin-resistant Staphylococcus aureus (MRSA) strains from cattle and human and to determine their genetic relatedness comparing the DNA restriction patterns. METHODS: Strains of Staphylococcus aureus were isolated from animals (223 strains) and humans (83). The E-test was applied to determine methicillin-resistance. The restriction patterns of DNA were carried out with pulsed field gel electrophoresis (PFGE). RESULTS: Thirty two (14.34%) from animals and 53 (63.8%) from men strains of S. aureus showed resistance to methicillin. PFGE demonstrated that the strains from human and veterinary pathology are different. The microrganisms isolated from men revealed, among them, an high similarity while only two strains, from animals, were considered identical. CONCLUSIONS: The resistance to methicillin involved both human and veterinary pathology. The human MRSA strains were higher than the animals ones. The strains isolated from animals showed a large genomic variability while in man the number of indistinguishable microrganisms, induces to suppose the existence of a prevalent clone. PFGE could be considered the gold standard for molecular characterisation of MRSA isolates.

Detection of Streptococcus dysgalactiae subsp. equisimilis in equine nasopharyngeal swabs by PCR.

Streptococcus (S.) dysgalactiae subsp. equisimilis is responsible for severe diseases in humans, including primary bacteraemia, pneumonia, endocarditis, and toxic shock syndrome. Infection in some animal species can also occur, although a few studies have looked into cross-species infectivity. In horses, S. equisimilis is generally considered infrequent or opportunistic, but has recently been isolated from cases of strangles-like disease. Rapid and sensitive diagnostic techniques could enable epidemiological studies and effective investigation of outbreaks involving these bacteria. In this study, PCR protocols previously described in cattle and in humans to detect the species S. dysgalactiae and the subspecies equisimilis were evaluated to detect specific sequences in equine samples. For this purpose, 99 monolateral nasal swabs were collected from horses from stud farms with a history of S. equisimilis infection and were tested blindly by bacteriological isolation and by single and duplex PCR. DNA for PCR was extracted both from the colonies grown on agar media and from enrichment broth aliquots after incubation with nasal swab samples. S. equisimilis was identified by bacteriological isolation in 23 out of 99 swab samples, and PCR assays on these colonies were fully concordant with bacteriological identification (kappa statistic = 1.00). In addition, PCR of the enrichment broth aliquots confirmed the bacteriological results and detected S. equisimilis in 6 samples more than the bacteriological examination (kappa statistic = 0.84). The PCR protocols appeared to be reliable for the rapid identification of S. equisimilis in equine nasal swab samples, and could be useful for microbiological diagnosis.

Attenuated Salmonella enterica serovar Typhimurium lacking the ZnuABC transporter confers immune-based protection against challenge infections in mice.

Salmonella enterica has long been recognised as an important zoonotic pathogen of economic significance, both in animals and humans. We have recently shown that inactivation of the ZnuABC high affinity zinc transporter significantly affects the pathogenicity of S. enterica, likely due to zinc shortage in the eukaryotic tissues. Here, we demonstrate that a S. enterica serovar Typhimurium znuABC deleted strain is able to induce a short lasting infection in mice. On the same time, it primes a cell-mediated immune response, which confers a solid and durable immune-based protection against challenge infections with virulent strains of S. Typhimurium. These findings suggest the possibility to explore the use of S. enterica ZnuABC deleted mutants for the production on novel vaccines.

Cloning, expression, and functional characterization of the equine herpesvirus 1 DNA polymerase and its accessory subunit.

We report the expression and characterization of the putative catalytic subunit (pORF30) and accessory protein (pORF18) of equine herpesvirus 1 DNA polymerase, which are encoded by open reading frames 30 and 18 and are homologous to herpes simplex virus type 1 UL30 and UL42, respectively. In vitro transcription-translation of open reading frames 30 and 18 generated proteins of 136 and 45 kDa, respectively. In vitro-expressed pORF30 possessed basal DNA polymerase activity that was stimulated by pORF18, as measured by DNA polymerase assays in vitro. Purified baculovirus-expressed pORF30 exhibited DNA polymerase activity similar to that of the in vitro-expressed protein, and baculovirus-expressed pORF18 could stimulate both nucleotide incorporation and long-chain DNA synthesis by pORF30 in a dose- and time-dependent manner. The salt optima for activity of both pORF30 and the holoenzyme were substantially different from those for other herpesvirus DNA polymerases. As demonstrated by yeast two-hybrid assays, pORF30 and pORF18 could physically interact, most likely with a 1:1 stoichiometry. Finally, by mutational analysis of the 1,220-residue pORF30, we demonstrated that the extreme C terminus of pORF30 is important for physical and functional interaction with the accessory protein, as reported for UL30 and other herpesvirus DNA polymerases. In addition, a C-proximal region of pORF30, corresponding to residues 1114 to 1172, is involved in binding to, and stimulation by, pORF18. Taken together, the results indicate that pORF30 and pORF18 are the equine herpesvirus 1 counterparts of herpes simplex virus type 1 UL30 and UL42 and share many, but not all, of their characteristics.