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1.
Perez-Pineiro R Bjorndahl TC Berjanskii MV Hau D Li L Huang A Lee R Gibbs E Ladner C Dong YW Abera A Cashman NR Wishart DS 《The FEBS journal》2011,278(21):4002-4014
Although highly conserved throughout evolution, the exact biological function of the prion protein is still unclear. In an effort to identify the potential biological functions of the prion protein we conducted a small-molecule screening assay using the Syrian hamster prion protein [shPrP(90-232)]. The screen was performed using a library of 149 water-soluble metabolites that are known to pass through the blood-brain barrier. Using a combination of 1D NMR, fluorescence quenching and surface plasmon resonance we identified thiamine (vitamin B1) as a specific prion ligand with a binding constant of ~60 μM. Subsequent studies showed that this interaction is evolutionarily conserved, with similar binding constants being seen for mouse, hamster and human prions. Various protein construct lengths, both with and without the unstructured N-terminal region in the presence and absence of copper, were examined. This indicates that the N-terminus has no influence on the protein's ability to interact with thiamine. In addition to thiamine, the more biologically abundant forms of vitamin B1 (thiamine monophosphate and thiamine diphosphate) were also found to bind the prion protein with similar affinity. Heteronuclear NMR experiments were used to determine thiamine's interaction site, which is located between helix 1 and the preceding loop. These data, in conjunction with computer-aided docking and molecular dynamics, were used to model the thiamine-binding pharmacophore and a comparison with other thiamine binding proteins was performed to reveal the common features of interaction. 相似文献
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VR Chowdhary AY Tilahun CR Clark JP Grande G Rajagopalan 《Journal of immunology (Baltimore, Md. : 1950)》2012,189(4):2054-2062
Chronic nasal and skin colonization with superantigen (SAg)-producing Staphylococcus aureus is well documented in humans. Given that trans-mucosal and trans-cutaneous absorption of SAgs can occur, we determined whether chronic exposure to small amounts of SAg per se could activate autoreactive CD4(+) and CD8(+) T cells and precipitate any autoimmune disease without further external autoantigenic stimulation. Because HLA class II molecules present SAg more efficiently than do mouse MHC class II molecules, HLA-DQ8 transgenic mice were implanted s.c. with mini-osmotic pumps capable of continuously delivering the SAg, staphylococcal enterotoxin B (total of 10 μg/mouse), or PBS over 4 wk. Chronic exposure to staphylococcal enterotoxin B resulted in a multisystem autoimmune inflammatory disease with features similar to systemic lupus erythematosus. The disease was characterized by mononuclear cell infiltration of lungs, liver, and kidneys, accompanied by the production of anti-nuclear Abs and deposition of immune complexes in the renal glomeruli. The inflammatory infiltrates in various organs predominantly consisted of CD4(+) T cells bearing TCR Vβ8. The extent of immunopathology was markedly reduced in mice lacking CD4(+) T cells and CD28, indicating that the disease is CD4(+) T cell mediated and CD28 dependent. The absence of disease in STAT4-deficient, as well as IFN-γ-deficient, HLA-DQ8 mice suggested the pathogenic role of Th1-type cytokines, IL-12 and IFN-γ. In conclusion, our study suggests that chronic exposure to extremely small amounts of bacterial SAg could be an etiological factor for systemic lupus erythematosus. 相似文献
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Eden Ephraim Agnes Odenyo Mogessie Ashenafi 《World journal of microbiology & biotechnology》2005,21(6-7):803-809
Summary Tannins limit the use of fodder trees as feed for ruminants. Removal of the effects of tannins would thus improve the nutritional
quality of these trees. This prompted the study to evaluate the effect of rumen or faecal mixed cultures from different animals
on tannin degradation. Tannin extracts, tannic acid and gallic acid were used to enrich media to assess if rumen or faecal
mixed cultures could degrade the phenolic compounds. Rumen fluid of Acacia-adapted sheep, sheep fed on wheat bran, bush duikers (Sylvicapra grimmia) and goats fed on Leucaena pallida and Sesbania goetzei were separately inoculated into Growth Study Medium (GSM) and incubated for 5-15 days. Faecal samples from dikdik (Madoqua guentheri), camel (Camelus dromedarius), zebra (Equus quagga), Grant’s gazelle (Gazella granti) and hartebeest (Alcelaphus buselaphus) were also separately inoculated into GSM media and incubated from 3-5 days. TLC results showed that mixed cultures from
rumen fluids of Acacia-adapted sheep, sheep on wheat bran, goats on Leucaena pallida and Sesbania goetzei partially hydrolysed tannic acid to pyrogallol. Complete degradation of the heterocyclic ring in tannic acid and gallic acid
was achieved by the mixed cultures from the faecal samples of dikdik and this was confirmed by HPLC. Mixed cultures from faecal
samples of camel hydrolysed gallic acid to phloroglucinol. This study has demonstrated that faecal microorganisms of Ethiopian
dikdik could completely degrade hydrolysable tannin. 相似文献
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Takele Beyene Tufa Feraol Gurmu Ashenafi Feyisa Beyi Henk Hogeveen Tariku Jibat Beyene Dinka Ayana Fanos Tadesse Woldemariyam Eyerusalem Hailemariam Fanta Desissa Gutema J. A. Stegeman 《BMC veterinary research》2018,14(1):409
Background
Antimicrobials and anthelmintics are the most commonly used veterinary drugs to control animal diseases. However, widespread use of these drugs could contribute to the emergence of drug resistance. Information on the practice of antimicrobial usage among food animal raising communities in Central Ethiopia is scarce. We used a standardised questionnaire survey to assess knowledge, awareness, and practices related to drug use and resistance in food animals among the farmers in and around Bishoftu town.Results
Of the total of 220 livestock owners interviewed, around 80% of the respondents were not able to define what antimicrobials are and for what purposes they are used. Only 14.1% (n?=?31) of the respondents had awareness about antimicrobial resistance (AMR) and its consequences; and 35.5% (n?=?11/31) and 9.7% (n?=?3/31) of them agreed that the irrational use of antimicrobials in animals could lead to AMR in animals and humans. Oxytetracycline was the most commonly available antibiotic in veterinary drug shops/pharmacies and the most widely used drug in the area. However, 43.3% of the respondents did not see clinical improvements after using antibiotics. Similarly, the respondents explained that no response was observed in 73.3, 70.8 and 52.5% of the cases after medication with anthelmintics, antiprotozoal and acaricides, respectively. About 56.7% of the respondents considered traditional medicines equally important to modern medicines. It was also noted that there were illegal drug vendors, dispensing medicines under unfavourable conditions which include a direct exposure to sunlight, which practice violates the drug handling and storage recommendations given by WHO.Conclusion
The study revealed that there is a general lack of awareness among food animal owners about the correct use of antibiotics and anthelmintics. The widespread misuse and improper drug dispensing and handling practices observed in this study can affect the drug quality and can also contribute to the development of drug resistance in central Ethiopia.6.
M. Ashenafi 《World journal of microbiology & biotechnology》1989,5(3):313-319
Summary A total of 708 strains belonging to different bacterial genera were isolated from fresh tomatoes, eggs, green peppers and fish sold in open markets in Awassa, Ethiopia. They mainly consisted of coliforms (54%),Staphylococcus aureus (26%) andProteus (16%).Shigella was isolated from 26% of the tomato specimens. Total counts for the various food items varied between 2.4 × 106 c.f.u./g and 9.5 × 106 c.f.u./g. Most probable number of total coliform and faecal coliform ranged from 1.5 × 102/g to 3 × 102/g and 37/g to 70/g. Less than 50% of the isolated 522 Gram-negative rods were susceptible to ampicillin and cephalothin and only 14 isolates were susceptible to all drugs tested.Proteus showed higher rates of resistance to ampicillin, carbenicillin and cephalothin (75%). Resistance to these drugs was also relatively higher inEscherichia coli (50%). Sixty-eight of the 186S. aureus isolates were sensitive to all the drugs tested and susceptibility to tetracycline and chloramphenicol was relatively lower (70%).
Charge microbienne, incidence et résistance aux antibiotiques de quelques micro-organismes dans des aliments destinés à être consommés crus en Ethiopie
Résumé On a isolé un total de 708 souches appartenant à divers genres bactériens de tomates fraîches, d'oeufs, de poivrons verts et de poissons vendus sur les marchés ouverts en Awassa, Ethiopie. Parmi celles-ci, on a trouvé 26% deStaphylococcus aureus et 16% deProteus. On a isolé desShigella dans 26% des tomates testées. L'énumération totale des germes dans divers aliments a donné des valeurs situées entre 2.4 × 106 c.f.u./g et 9.5 × 106 c.f.u./g. Le nombre le plus probable de coliformes totaux et de coliformes fécaux variait entre 1.5 × 102/g et 3 × 102/g et entre 37/g et 70/g. Moins de 50% des 522 souches isolées à forme de bâtonnets étaient sensibles à l'ampicilline et à la céphalothine et seules 14 souches isolées étaient sensibles à toutes les drogues testées. LesProteus ont montré davantage de résistance à l'ampicilline, la carbénicilline et la céphalothine (75%). La résistance à ces drogues était aussi relativement plus élevée chezEscherichia coli. Soixante huit de 186 souches isolées deS. aureus étaient sensibles à toutes les drogues testées et leur sensibilité à la tétracycline et au chloramphenicol était relativement plus faible (70%).相似文献
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Counts of Bacillus cereus reached ca 108 cfu/g within 40 h in fermenting unacidified horsebean tempeh and resulted in complete spoilage of the product. In fermenting unacidified pea, chickpea and soybean tempeh, B. cereus counts reached 106 –107 cfu/g, although the products were not spoiled. Inoculation of these unacidified beans with Lactobacillus plantarum decreased the final count of B. cereus by 2 log units, but had no effect on its growth in unacidified horsebean tempeh and its subsequent spoilage. Acidification of the beans during soaking resulted in a lower rate of B. cereus growth during fermentation. Inoculation of acidified beans with Lact. plantarum resulted in a markedly lower growth rate of B. cereus . In an associative broth culture study, B. cereus was completely inhibited by Lact. plantarum at pH values of about 5·5. Lactobacillus plantarum may be used to control the growth of B. cereus during tempeh production. 相似文献
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Endalkachew Ashenafi Alemu Trond Lamark Knut Martin Torgersen Aasa Birna Birgisdottir Kenneth Bowitz Larsen Ashish Jain Hallvard Olsvik Aud ?vervatn Vladimir Kirkin Terje Johansen 《The Journal of biological chemistry》2012,287(47):39275-39290
Autophagy is a lysosome-dependent degradation system conserved among eukaryotes. The mammalian Atg1 homologues, Unc-51 like kinase (ULK) 1 and 2, are multifunctional proteins with roles in autophagy, neurite outgrowth, and vesicle transport. The mammalian ULK complex involved in autophagy consists of ULK1, ULK2, ATG13, FIP200, and ATG101. We have used pulldown and peptide array overlay assays to study interactions between the ULK complex and six different ATG8 family proteins. Strikingly, in addition to ULK1 and ULK2, ATG13 and FIP200 interacted with human ATG8 proteins, all with strong preference for the GABARAP subfamily. Similarly, yeast and Drosophila Atg1 interacted with their respective Atg8 proteins, demonstrating the evolutionary conservation of the interaction. Use of peptide arrays allowed precise mapping of the functional LIR motifs, and two-dimensional scans of the ULK1 and ATG13 LIR motifs revealed which substitutions that were tolerated. This information, combined with an analysis of known LIR motifs, provides us with a clearer picture of sequence requirements for LIR motifs. In addition to the known requirements of the aromatic and hydrophobic residues of the core motif, we found the interactions to depend strongly on acidic residues surrounding the central core LIR motifs. A preference for either a hydrophobic residue or an acidic residue following the aromatic residue in the LIR motif is also evident. Importantly, the LIR motif is required for starvation-induced association of ULK1 with autophagosomes. Our data suggest that ATG8 proteins act as scaffolds for assembly of the ULK complex at the phagophore. 相似文献
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