69 research outputs found
Proline-Rich Peptides with Improved Antimicrobial Activity against E. coli, K. Pneumoniae, and A. Baumannii.
No full textProline-rich antimicrobial peptides (PrAMPs) are promising agents to combat multi-drug resistant pathogens due to a high antimicrobial activity, yet low cytotoxicity. A library of derivatives of the PrAMP Bac5(1-17) was synthesized and screened to identify which residues are relevant for its activity. In this way, we discovered that two central motifs -PIRXP- cannot be modified, while residues at N- and C- termini tolerated some variations. We found five Bac5(1-17) derivatives bearing 1-5 substitutions, with an increased number of arginine and/or tryptophan residues, exhibiting improved antimicrobial activity and broader spectrum of activity while retaining low cytotoxicity toward eukaryotic cells. Transcription/translation and bacterial membrane permeabilization assays showed that these new derivatives still retained the ability to strongly inhibit bacterial protein synthesis, but also acquired permeabilizing activity to different degrees. These new Bac5(1-17) derivatives therefore show a dual mode of action which could hinder the selection of bacterial resistance against these molecules
Peptide inhibitors of bacterial protein synthesis with broad spectrum and SbmA-independent bactericidal activity against clinical pathogens.
No full textProline-rich antimicrobial peptides (PrAMPs) are promising lead compounds for developing new antimicrobials, however their narrow spectrum of action is limiting. PrAMPs kill bacteria binding to their ribosomes and inhibiting protein synthesis. In this study, 133 derivatives of the PrAMP Bac7(1-16) were synthesized to identify the crucial residues for ribosome inactivation and antimicrobial activity. Then, five new Bac7(1-16) derivatives were conceived and characterized by antibacterial and membrane permeabilization assays, by X-ray crystallography and molecular dynamics simulations. Some derivatives displayed broad spectrum activity, encompassing Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa and Staphylococcus aureus. Two peptides out of five, acquired a weak membrane-perturbing activity, while maintaining the ability to inhibit protein synthesis. These derivatives became independent of the SbmA transporter, commonly used by native PrAMPs, suggesting that they obtained a novel route to enter bacterial cells. PrAMP-derived compounds could become new-generation antimicrobials to combat the antibiotic-resistant pathogens
The effect of lipidation and glycosylation on short cationic antimicrobial peptides
No full textThe global health threat surrounding bacterial resistance has resulted in antibiotic researchers shifting their focus away from ‘traditional’ antibiotics and concentrating on other antimicrobial agents, including antimicrobial peptides. These low molecular weight "mini-proteins" exhibit broad-spectrum activity against bacteria, including multi-drug resistant strains, viruses, fungi and protozoa and constitute a major element of the innate-immune system of many multicellular organisms. Some naturally occurring antimicrobial peptides are lipidated and/or glycosylated and almost all antimicrobial peptides in clinical use are either lipopeptides (Daptomycin and Polymyxin E and B) or glycopeptides (Vancomycin). Lipidation, glycosylation and PEGylation are an option for improving stability and activity in serum and for reducing the rapid clearing via the kidneys and liver. Two broad-spectrum antimicrobial peptides NH2-RIRIRWIIR-CONH2 (A1) and NH2-KRRVRWIIW-CONH2 (B1) were conjugated via a linker, producing A2 and B2, to individual fatty acids of C8, C10, C12 and C14 and in addition, A2 was conjugated to either glucose, N-acetyl glucosamine, galactose, mannose, lactose or polyethylene glycol (PEG). Antimicrobial activity against two Gram-positive strains (methicillin resistant Staphylococcus aureus (MRSA) and vancomycin resistant Enterococcus faecalis (VRE)) and three Gram-negative strains (Salmonella typhimurium, E. coli and Pseudomonas aeruginosa) were determined. Activity patterns for the lipidated versions are very complex, dependent on sequence, bacteria and fatty acid. Two reciprocal effects were measured; compared to the parental peptides, some combinations led to a 16-fold improvement whereas other combinations let to a 32-fold reduction in antimicrobial activity. Glycosylation decreased antimicrobial activity by 2 to 16-fold in comparison to A1, respectively on the sugar-peptide combination. PEGylation rendered the peptide inactive. Antimicrobial activity in the presence of 25% human serum of A1 and B1 was reduced 32-fold and 8-fold, respectively. The longer chain fatty acids almost completely restored this activity; however, these fatty acids increased hemolytic activity. B1 modified with C8 increased the therapeutic index by 2-fold for four bacterial strains. Our results suggest that finding the right lipid-peptide combination can lead to improved activity in the presence of serum and potentially more effective drug candidates for animal studies. Glycosylation with the optimal sugar and numbers of sugars at the right peptide position could be an alternative route or could be used in addition to lipidation to counteract solubility and toxicity issues
Database-driven activities to support web-based learning
No full textThis paper describes and rationalises the use of database driven activities and resources for Web-based learning. Database driven Web resources differ from conventional Web-based resources in that they tend not to exist in discrete forms of HTML but are generated dynamically through such means as learner interaction. Database driven applications are built around the use of the Web as' a means to store, retrieve and develop media resource. The applications are controlled by software engines that provide the functionality and flexibility for both teachers' and students' to interact with Web resources and content. This' paper describes various forms of database driven resources for learners' and discusses the learning opportunities that result
New technologies, new pedagogies: Mobile technologies and new ways of teaching and learning
No full textThis paper describes a major development and research study that investigated the use of mobile technologies in higher education. The project investigated the educational potential of two ubiquitous mobile devices: Palm smart phones and iPod digital audio players (mp3 players). An action learning framework for professional development was designed and implemented with a group of teachers from a Faculty of Education. Each teacher or team created pedagogies to implement appropriate use of a mobile device in different subject areas in higher education. This paper describes the project aims, design and implementation in four phases, together with a description of the project management and communication factors that helped to ensure its success
Learning from biophysical heterogeneity: inductive use of case studies for maize cropping systems in Central America
No full textGlobal society has become conscious that efforts towards securing food production will only be successful if agricultural production increases are obtained through mechanisms that ensure active regeneration of the natural resource base. Production options should be targeted in the sense of that their suitability to improve agricultural production and maintain natural resources is evaluated prior to their introduction. Biophysical targeting evaluates production options as a function of the spatial and temporal variability of climate conditions, in interaction with soil, crop characteristics and agronomic management strategies. This thesis contributes to the development of a system-based methodology for biophysical targeting. Cropping system simulation and weather generator tools are interfaced to geographical information systems. Inductive use of two case studies - a green manure cover crop and reduced tillage with residue management - helped to develop the methodology. Insight is gained into the regional potential for and the soil and climate conditions under which successful introduction of these production options may be achieved. The resulting information supports regional stakeholders involved in agriculture in their analysis and discussion, negotiation and decision-making concerning where to implement production systems. This process can improve the supply of appropriate agricultural production practices that enhance production and conserve soil and water resources
Supramolecular Presentation of Hyaluronan onto Model Surfaces for Studying the Behaviour of Cancer Stem Cells
No full textThe supramolecular presentation of extracellular matrix components on surfaces provides a platform for the investigation and control of cell behavior. Hyaluronan (HA) is one of the main components of the extracellular environment and has been shown to play an important role in different cancers and their progression. However, current methods of HA immobilization often require its chemical modification. Herein, a peptide-based self-assembled monolayer (SAM) is used as an anchor to immobilize unmodified HA on a bare gold surface, as demonstrated by the quartz crystal microbalance with dissipation monitoring. Peptide-HA surfaces show increased roughness and greater hydrophobicity when compared to poly-D-lysine/HA surfaces, as measured by atomic force microscopy and water contact angle, respectively. Additionally, the peptide SAM can be micro-contact printed and used to restrict the presentation of HA to specific regions, thereby creating HA patterned surfaces to examine cell behavior. When used for cell culture, these surfaces result in altered adhesion and migration of LUC4 head and neck squamous cell carcinoma cells. These biomimetic surfaces can provide insights into the role of HA in cancer and other diseases and be used as a platform for the development of cell sorting devices.C.O’M. thanks The Queen Mary Institute of Bioengineering and the
Engineering and Physical Sciences Research Council for financial support through a Ph.D. studentship (Award number 1502316).
J.B. gratefully acknowledges the financial support by Fundação para
a Ciência e a Tecnologia (FCT), I.P., through individual contract
(CEECIND/03202/2017). This work was also funded by national
funds (OE), through FCT, I.P., in the scope of the framework contract
foreseen in the numbers 4, 5, and 6 of the article 23, of the Decree-Law
57/2016, of August 29, changed by Law 57/2017, of July 19. The authors
also acknowledge the financial support from the EU-funded project
“SuprHApolymers” (PCIG14-GA-2013-631871).publishe
The design dimensions of e-learning
No full textThe currently popular term e-learning is shown to have several meanings which confuse discussion about a new technology which is finding widespread international use in a range of educational sectors. This paper analyses the characteristics of e-learning applications, and proposes a set of four design dimensions which could be considered when designing and assessing the suitability of e-learning applications. The applicability of the four proposed dimensions, and their shortcomings, are discussed in detail, and found to be justifiable
Can BioSAXS detect ultrastructural changes of antifungal compounds in Candida albicans?–an exploratory study
No full textThe opportunistic yeast Candida albicans is the most common cause of candidiasis. With only four classes of antifungal drugs on the market, resistance is becoming a problem in the treatment of fungal infections, especially in immunocompromised patients. The development of novel antifungal drugs with different modes of action is urgent. In 2016, we developed a groundbreaking new medium-throughput method to distinguish the effects of antibacterial agents. Using small-angle X-ray scattering for biological samples (BioSAXS), it is now possible to screen hundreds of new antibacterial compounds and select those with the highest probability for a novel mode of action. However, yeast (eukaryotic) cells are highly structured compared to bacteria. The fundamental question to answer was if the ultrastructural changes induced by the action of an antifungal drug can be detected even when most structures in the cell stay unchanged. In this exploratory work, BioSAXS was used to measure the ultrastructural changes of C. albicans that were directly or indirectly induced by antifungal compounds. For this, the well-characterized antifungal drug Flucytosine was used. BioSAXS measurements were performed on the synchrotron P12 BioSAXS beamline, EMBL (DESY, Hamburg) on treated and untreated yeast C. albicans. BioSAXS curves were analysed using principal component analysis (PCA). The PCA showed that Flucytosine-treated and untreated yeast were separated. Based on that success further measurements were performed on five antifungal peptides {1. Cecropin A-melittin hybrid [CA (1–7) M (2–9)], KWKLFKKIGAVLKVL; 2. Lasioglossin LL-III, VNWKKILGKIIKVVK; 3. Mastoparan M, INLKAIAALAKKLL; 4. Bmkn2, FIGAIARLLSKIFGKR; and 5. optP7, KRRVRWIIW}. The ultrastructural changes of C. albicans indicate that the peptides may have different modes of action compared to Flucytosine as well as to each other, except for the Cecropin A-melittin hybrid [CA (1–7) M (2–9)] and optP7, showing very similar effects on C. albicans. This very first study demonstrates that BioSAXS shows promise to be used for antifungal drug development. However, this first study has limitations and further experiments are necessary to establish this application
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