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101.
Wendy E. Kaman Ingrid Voskamp-Visser Denise M.C. de Jongh Hubert P. Endtz Alex van Belkum John P. Hays Floris J. Bikker 《Analytical biochemistry》2013
Bacterial proteases play an important role in a broad spectrum of processes, including colonization, proliferation, and virulence. In this respect, bacterial proteases are potential biomarkers for bacterial diagnosis and targets for novel therapeutic protease inhibitors. To investigate these potential functions, the authors designed and used a protease substrate fluorescence resonance energy transfer (FRET) library comprising 115 short d- and l-amino-acid-containing fluorogenic substrates as a tool to generate proteolytic profiles for a wide range of bacteria. Bacterial specificity of the d-amino acid substrates was confirmed using enzymes isolated from both eukaryotic and prokaryotic organisms. Interestingly, bacterial proteases that are known to be involved in housekeeping and nutrition, but not in virulence, were able to degrade substrates in which a d-amino acid was present. Using our FRET peptide library and culture supernatants from a total of 60 different bacterial species revealed novel, bacteria-specific, proteolytic profiles, although in-species variation was observed for Pseudomonas aeruginosa, Porphyromonas gingivalis, and Staphylococcus aureus. Overall, the specific characteristic of our substrate peptide library makes it a rapid tool to high-throughput screen for novel substrates to detect bacterial proteolytic activity. 相似文献
102.
Ligiane R. Gouvea Darliane A. Martins Denise da Gama Jean Batista Maria de Nazaré C. Soeiro Sonia R. W. Louro Paulo J. S. Barbeira Letícia R. Teixeira 《Biometals》2013,26(5):813-825
Zn(II) complexes with norfloxacin (NOR) in the absence or in the presence of 1,10-phenanthroline (phen) were obtained and characterized. In both complexes, the ligand NOR was coordinated through a keto and a carboxyl oxygen. Tetrahedral and octahedral geometries were proposed for [ZnCl2(NOR)]·H2O (1) and [ZnCl2(NOR)(phen)]·2H2O (2), respectively. Since the biological activity of the chemicals depends on the pH value, pH titrations of the Zn(II) complexes were performed. UV spectroscopic studies of the interaction of the complexes with calf-thymus DNA (CT DNA) have suggested that they can bind to CT DNA with moderate affinity in an intercalative mode. The interactions between the Zn(II) complexes and bovine serum albumin (BSA) were investigated by steady-state and time-resolved fluorescence spectroscopy at pH 7.4. The experimental data showed static quenching of BSA fluorescence, indicating that both complexes bind to BSA. A modified Stern–Volmer plot for the quenching by complex 2 demonstrated preferential binding near one of the two tryptophan residues of BSA. The binding constants obtained (K b ) showed that BSA had a two orders of magnitude higher affinity for complex 2 than for 1. The results also showed that the affinity of both complexes for BSA was much higher than for DNA. This preferential interaction with protein sites could be important to their biological mechanisms of action. The analysis in vitro of the Zn(II) complexes and corresponding ligand were assayed against Trypanosoma cruzi, the causative agent of Chagas disease and the data showed that complex 2 was the most active against bloodstream trypomastigotes. 相似文献
103.
AbstractAn evaluation of nitrite determination in marine lake sediments has shown that spectrophotometric measurements can be in error due to light scattering by colloidal (<0.2 μm) matter in extract solutions and incomplete nitrite recovery. The scatter error can be minimised by using uncoloured extract in the reference beam but precision at low levels remains poor (RSD 25 to 100%). Recovery tests on ‘spiked’ sediment indicated that optimum retrieval (~85%) occurred with 30 minute mixing with 0.2 M NH4Cl, using a sediment to extractant ratio of 1:30. To counter this variable, calibration based on standard addition to sample suspensions is recommended. Modified procedure proposed is suitable for measuring up to 10 μg g?1 of nitrite N; the lake sediments tested contained <100 ng g?1 相似文献
104.
Ronan A. Lyons Denise Kendrick Elizabeth M. L. Towner Carol Coupland Mike Hayes Nicola Christie Judith Sleney Sarah Jones Richard Kimberlee Sarah E. Rodgers Samantha Turner Mariana Brussoni Yana Vinogradova Tinnu Sarvotham Steven Macey 《PloS one》2013,8(4)
Objective
To determine whether advocacy targeted at local politicians leads to action to reduce the risk of pedestrian injury in deprived areas.Design
Cluster randomised controlled trial.Setting
239 electoral wards in 57 local authorities in England and Wales.Participants
617 elected local politicians.Interventions
Intervention group politicians were provided with tailored information packs, including maps of casualty sites, numbers injured and a synopsis of effective interventions.Main outcome measures
25–30 months post intervention, primary outcomes included: electoral ward level: percentage of road traffic calmed; proportion with new interventions; school level: percentage with 20 mph zones, Safe Routes to School, pedestrian training or road safety education; politician level: percentage lobbying for safety measures. Secondary outcomes included politicians’ interest and involvement in injury prevention, and facilitators and barriers to implementation.Results
Primary outcomes did not significantly differ: % difference in traffic calming (0.07, 95%CI: −0.07 to 0.20); proportion of schools with 20 mph zones (RR 1.47, 95%CI: 0.93 to 2.32), Safe Routes to School (RR 1.34, 95%CI: 0.83 to 2.17), pedestrian training (RR 1.23, 95%CI: 0.95 to 1.61) or other safety education (RR 1.16, 95%CI: 0.97 to 1.39). Intervention group politicians reported greater interest in child injury prevention (RR 1.09, 95%CI 1.03 to 1.16), belief in potential to help prevent injuries (RR 1.36, 95%CI 1.16 to 1.61), particularly pedestrian safety (RR 1.55, 95%CI 1.19 to 2.03). 63% of intervention politicians reported supporting new pedestrian safety schemes. The majority found the advocacy information surprising, interesting, effectively presented, and could identify suitable local interventions.Conclusions
This study demonstrates the feasibility of an innovative approach to translational public health by targeting local politicians in a randomised controlled trial. The intervention package was positively viewed and raised interest but changes in interventions were not statistically significance. Longer term supported advocacy may be needed.Trial Registration
Current Controlled Trials ISRCTN91381117 相似文献105.
Britta Stadelmann Reto Rufener Denise Aeschbacher Markus Spiliotis Bruno Gottstein Andrew Hemphill 《PLoS neglected tropical diseases》2016,10(3)
The metacestode (larval) stage of the tapeworm Echinococcus multilocularis causes alveolar echinococcosis (AE), a very severe and in many cases incurable disease. To date, benzimidazoles such as albendazole and mebendazole are the only approved chemotherapeutical treatment options. Benzimidazoles inhibit metacestode proliferation, but do not act parasiticidal. Thus, benzimidazoles have to be taken a lifelong, can cause adverse side effects such as hepatotoxicity, and are ineffective in some patients. We here describe a newly developed screening cascade for the evaluation of the in vitro efficacy of new compounds that includes assessment of parasiticidal activity. The Malaria Box from Medicines for Malaria Venture (MMV), comprised of 400 commercially available chemicals that show in vitro activity against Plasmodium falciparum, was repurposed. Primary screening was carried out at 10 μM by employing the previously described PGI assay, and resulted in the identification of 24 compounds that caused physical damage in metacestodes. Seven out of these 24 drugs were also active at 1 μM. Dose-response assays revealed that only 2 compounds, namely MMV665807 and MMV665794, exhibited an EC50 value below 5 μM. Assessments using human foreskin fibroblasts and Reuber rat hepatoma cells showed that the salicylanilide MMV665807 was less toxic for these two mammalian cell lines than for metacestodes. The parasiticidal activity of MMV665807 was then confirmed using isolated germinal layer cell cultures as well as metacestode vesicles by employing viability assays, and its effect on metacestodes was morphologically evaluated by electron microscopy. However, both oral and intraperitoneal application of MMV665807 to mice experimentally infected with E. multilocularis metacestodes did not result in any reduction of the parasite load. 相似文献
106.
107.
Reduced hnRNPA3 increases C9orf72 repeat RNA levels and dipeptide‐repeat protein deposition
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Kohji Mori Yoshihiro Nihei Thomas Arzberger Qihui Zhou Ian R Mackenzie Andreas Hermann Frank Hanisch Frits Kamp Brigitte Nuscher Denise Orozco Dieter Edbauer Christian Haass 《EMBO reports》2016,17(9):1314-1325
Intronic hexanucleotide (G4C2) repeat expansions in C9orf72 are genetically associated with frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). The repeat RNA accumulates within RNA foci but is also translated into disease characterizing dipeptide repeat proteins (DPR). Repeat‐dependent toxicity may affect nuclear import. hnRNPA3 is a heterogeneous nuclear ribonucleoprotein, which specifically binds to the G4C2 repeat RNA. We now report that a reduction of nuclear hnRNPA3 leads to an increase of the repeat RNA as well as DPR production and deposition in primary neurons and a novel tissue culture model that reproduces features of the C9orf72 pathology. In fibroblasts derived from patients carrying extended C9orf72 repeats, nuclear RNA foci accumulated upon reduction of hnRNPA3. Neurons in the hippocampus of C9orf72 patients are frequently devoid of hnRNPA3. Reduced nuclear hnRNPA3 in the hippocampus of patients with extended C9orf72 repeats correlates with increased DPR deposition. Thus, reduced hnRNPA3 expression in C9orf72 cases leads to increased levels of the repeat RNA as well as enhanced production and deposition of DPR proteins and RNA foci. 相似文献
108.
109.
Changes in interspecies association patterns of Atlantic bottlenose dolphins,Tursiops truncatus,and Atlantic spotted dolphins,Stenella frontalis,after demographic changes related to environmental disturbance
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Animal populations can be affected by environmental disturbances in many ways including demographic and behavioral changes. This can affect interspecies associations for regularly interacting sympatric species, like bottlenose and spotted dolphins in the Bahamas (observed since 1985 and interspecies associations analyzed since 1993). After two hurricanes in 2004 each species lost roughly 30% of their respective communities resulting in differing social structure and behavioral changes. During mixed species encounters (MSE) group sizes for spotted dolphins ( = 14.1 ± 9.2) were significantly larger than bottlenose dolphins ( = 6.0 ± 7.3; F = 11.74, df = 1, P < 0.001), however, t‐tests revealed no differences between aggressive vs. affiliative encounters. Sexual/aggressive behavior regularly seen previously was not observed posthurricanes and aggressive encounters were greatly reduced. Generally results were similar to prehurricane data including high resightings of spotted dolphins with male alliances prevalent (including new juvenile alliances seen only posthurricane), and individualized bottlenose participation with few male alliances. However temporal associations varied compared to prehurricane. Interspecies association and behavior patterns were altered and likely affected by the changes in intraspecies association patterns following the hurricanes. However both species still participated in MSE, suggesting this is an important component of their ability to coexist as sympatric species. 相似文献
110.
Jonathan M. Large Kristian Birchall Nathalie S. Bouloc Andy T. Merritt Ela Smiljanic-Hurley Denise J. Tsagris Mary C. Wheldon Keith H. Ansell Peter J. Coombs Catherine A. Kettleborough David Whalley Lindsay B. Stewart Paul W. Bowyer David A. Baker Simon A. Osborne 《Bioorganic & medicinal chemistry letters》2019,29(3):509-514
Development of a class of bicyclic inhibitors of the Plasmodium falciparum cyclic GMP-dependent protein kinase (PfPKG), starting from known compounds with activity against a related parasite PKG orthologue, is reported. Examination of key sub-structural elements led to new compounds with good levels of inhibitory activity against the recombinant kinase and in vitro activity against the parasite. Key examples were shown to possess encouraging in vitro ADME properties, and computational analysis provided valuable insight into the origins of the observed activity profiles. 相似文献