RESUMO
Fracture-related infection (FRI) is a major complication in surgically fixed fractures. Instability of the fracture after fixation is considered a risk factor for infection; however, few experimental data are available confirming this belief. To study whether stable fractures led to higher infection clearance, mouse femoral osteotomies were fixed with either stable or unstable fixation and the surgical site was contaminated with either Staphylococcus epidermidis (S. epidermidis)or Staphylococcus aureus (S. aureus)clinical isolates. Infection progression was assessed at different time points by quantitative bacteriology, total cell counts in spleen and lymph node and histological analysis. Operated, non-inoculated mice were used as controls. Two inbred mouse strains (C57BL/6 and BALB/c) were included in the study to determine the influence of different host background in the outcome. Stable fixation allowed a higher proportion of C57BL/6 mice to clear S. epidermidis inoculation in comparison to unstable fixation. No difference associated with fixation type was observed for BALB/c mice. Inoculation with S. aureus resulted in a more severe infection for both stable and unstable fractures in both mouse strains; however, significant osteolysis around the screws rendered the stable group functionally unstable. Our results suggested that fracture stability could have an influence on S. epidermidis infection, although host factors also played a role. No differences were observed when using S. aureus, due to a more severe infection, leading to osteolysis and loss of stability in both groups. Further studies are required in order to address the biological features underlying the differences observed.
Assuntos
Fraturas do Fêmur/cirurgia , Fixação de Fratura/métodos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus epidermidis/crescimento & desenvolvimento , Animais , Carga Bacteriana , Biofilmes/crescimento & desenvolvimento , Feminino , Fraturas do Fêmur/microbiologia , Fixação de Fratura/efeitos adversos , Fixação de Fratura/instrumentação , Interações Hospedeiro-Patógeno , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Microscopia Eletrônica de Varredura , Osteólise/microbiologia , Especificidade da Espécie , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/fisiologia , Staphylococcus aureus/ultraestrutura , Staphylococcus epidermidis/fisiologia , Staphylococcus epidermidis/ultraestrutura , Infecção da Ferida Cirúrgica/etiologia , Infecção da Ferida Cirúrgica/microbiologiaRESUMO
Fifty Clostridium perfringens strains were isolated from individual dogs with acute diarrhoea that were not given antibiotics. Toxin types and minimal inhibitory concentrations of 15 antibiotics were determined for each of them. All strains harboured the alpha-toxin gene, 12 of them had both the alpha- and entero-toxin gene and 5 had both the alpha- and beta2-toxin gene. Eighteen percent of the isolates showed resistance to tetracycline and 54 % showed decreased susceptibility to metronidazole which is one of the most frequently used antibiotics in the treatment of canine diarrhoea. Apart from that, all isolates were susceptible to the remaining antibiotics tested. These findings lead to the conclusion that despite a general susceptibility to antibiotics in C. perfringens, resistance is developing in isolates from dogs. Therefore, careful identification of the pathogenic agent and antibiotic susceptibility testing should be performed prior to therapy in order to minimise further selection of antibiotic resistance.
Assuntos
Antibacterianos/farmacologia , Infecções por Clostridium/veterinária , Clostridium perfringens/efeitos dos fármacos , Doenças do Cão/microbiologia , Animais , Toxinas Bacterianas/genética , Proteínas de Ligação ao Cálcio/genética , Infecções por Clostridium/microbiologia , Clostridium perfringens/genética , Clostridium perfringens/isolamento & purificação , Cães , Resistência Microbiana a Medicamentos , Enterotoxinas/genética , Testes de Sensibilidade Microbiana/veterinária , Suíça , Fosfolipases Tipo C/genéticaRESUMO
Antioxidant activity of vegetable extracts is related to the nature and the amount of active components, mainly polyphenols; therefore, a correct quantification of these molecules should be required to define their concentration in such kind of vegetable extracts. A fast and accurate method to calculate molar absorption coefficients (epsilon), by using HPLC, has been tested on standard polyphenols and caffeine, and should be widely adapted for standardless quantitative analysis. Molar absorptivity (epsilon) of carnosic acid (CA) was determined from 200 to 300 nm, by the proposed method and those values were compared to tert-butyl-hydroxytoluene (BHT) ones for further comparative quantification.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Flavonoides/química , Fenóis/química , Espectrofotometria Ultravioleta/métodos , Abietanos , Diterpenos/química , Extratos Vegetais/química , PolifenóisRESUMO
To improve the malt rootlet value, the antioxidant potentialities of this byproduct of the malting industry have been analyzed. Three extracts have been considered from the points of view of dry matter yield, level of antioxidant compounds, and efficiency and cost of the extraction protocols. They respectively contain rootlet oil, free phenolic compounds, and bound phenolic compounds. The rootlet oil contains only a low quantity of tocopherols (respectively, 20.6 and 4.2 microgram of alpha-tocopherol and gamma-tocopherol per gram of dry rootlets), and a weak antioxidant activity, evaluated in a stripped corn oil by following spectrophotometrically the conjugated dienes, has been pointed out. The bound compound extract presents a good antioxidant power mainly due to the presence of trans-ferulic and trans-p-coumaric acids, but the dry matter yield is low (2%). The free compound extract has a good antioxidant power, and the valuable dry matter, mainly composed of proteins (52%), sugars (33%), and reducing compounds (5.5%), has a yield of 12%. The mixing of bound and free compound extracts presents an antagonistic effect on the antioxidant power, but a synergistic effect has been pointed out for the mixing of alpha-tocopherol and free compound extract.
Assuntos
Antioxidantes/farmacologia , Grão Comestível/química , Extratos Vegetais/farmacologia , Cromatografia Líquida de Alta Pressão , Óleo de Milho , Sinergismo Farmacológico , Hordeum , Fenóis/metabolismo , Raízes de Plantas/químicaAssuntos
Aciltransferases/análise , Mycobacterium/enzimologia , Acetilcoenzima A , Aciltransferases/metabolismo , Sulfato de Amônio , Autorradiografia , Soluções Tampão , Precipitação Química , Cromatografia em Gel , Cromatografia por Troca Iônica , Cromatografia em Camada Fina , Eletroforese em Gel de Poliacrilamida , Malonatos , Peso Molecular , Ácidos Palmíticos , Trítio , Ultracentrifugação , UltrassomRESUMO
Initiation factor eIF4G is an essential protein required for initiation of mRNA translation via the 5' cap-dependent pathway. It interacts with eIF4E (the mRNA 5' cap-binding protein) and serves as an anchor for the assembly of further initiation factors. With treatment of Saccharomyces cerevisiae with rapamycin or with entry of cells into the diauxic phase, eIF4G is rapidly degraded, whereas initiation factors eIF4E and eIF4A remain stable. We propose that nutritional deprivation or interruption of the TOR signal transduction pathway induces eIF4G degradation.
Assuntos
Fatores de Iniciação de Peptídeos/metabolismo , Polienos/farmacologia , Biossíntese de Proteínas , Saccharomyces cerevisiae/metabolismo , Transdução de Sinais , Ciclo Celular/efeitos dos fármacos , Fator de Iniciação 4A em Eucariotos , Fator de Iniciação 4E em Eucariotos , Fator de Iniciação Eucariótico 4G , Genótipo , Cinética , Fatores de Iniciação de Peptídeos/efeitos dos fármacos , RNA Mensageiro/metabolismo , Saccharomyces cerevisiae/citologia , Saccharomyces cerevisiae/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Sirolimo , Fatores de TempoRESUMO
In the yeast Saccharomyces cerevisiae a small protein named p20 is found associated with translation initiation factor eIF4E, the mRNA cap-binding protein. We demonstrate here that p20 is a repressor of cap-dependent translation initiation. p20 shows amino acid sequence homology to a region of eIF4G, the large subunit of the cap-binding protein complex eIF4F, which carries the binding site for eIF4E. Both, eIF4G and p20 bind to eIF4E and compete with each other for binding to eIF4E. The eIF4E-p20 complex can bind to the cap structure and inhibit cap-dependent but not cap-independent translation initiation: the translation of a mRNA with the 67 nucleotide omega sequence of tobacco mosaic virus in its 5' untranslated region (which was previously shown to render translation cap-independent) is not inhibited by p20. Whereas the translation of the same mRNA lacking the omega sequence is strongly inhibited by p20. Disruption of CAF20, the gene encoding p20, stimulates the growth of yeast cells, overexpression of p20 causes slower growth of yeast cells. These results show that p20 is a regulator of eIF4E activity which represses cap-dependent initiation of translation by interfering with the interaction of eIF4E with eIF4G, e.g. the formation of the eIF4F-complex.
Assuntos
Proteínas Fúngicas/metabolismo , Complexo Proteico Nuclear de Ligação ao Cap , Fatores de Iniciação de Peptídeos/metabolismo , Fosfoproteínas/metabolismo , Biossíntese de Proteínas , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/metabolismo , Sequência de Aminoácidos , Sítios de Ligação , Western Blotting , Divisão Celular , Eletroforese em Gel de Poliacrilamida , Fator de Iniciação 4E em Eucariotos , Fator de Iniciação 4F em Eucariotos , Fator de Iniciação Eucariótico 4G , Regulação Fúngica da Expressão Gênica , Dados de Sequência Molecular , Proteínas de Ligação a RNA/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Homologia de Sequência de AminoácidosRESUMO
The combination of reversed-phase high-performance liquid chromatography with coulometric detection allows the detection of phenolic antioxidants in complex matrices like plant extracts with a high degree of sensitivity and selectivity. According to their voltammetric behaviour, phenolic acids and tocopherols show maximal detector response at low potentials (100-450 mV) while flavonoids show optimal response at two different potential values (one at 0-300 mV and one at 600-900 mV). The potential corresponding to maximal detector response (MDRP) of phenolic acids was shown to be inversely proportional to their antioxidant efficiency as determined in a lipidic model system under strong oxidizing conditions (110 degrees C, intensive oxygenation) or by the DPPH() test. However, such a relationship was not observed for flavonoids.