183 research outputs found
Promising approaches to optimize the biological properties of the antimicrobial peptide esculentin-1a(1-21)NH2. amino acids substitution and conjugation to nanoparticles
Antimicrobial peptides (AMPs) represent an interesting class of molecules with expanding biological properties which make them a viable alternative for the development of future antibiotic drugs. However, for this purpose, some limitations must be overcome: (i) the poor biostability due to enzymatic degradation; (ii) the cytotoxicity at concentrations slightly higher than the therapeutic dosages; and (iii) the inefficient delivery to the target site at effective concentrations. Recently, a derivative of the frog skin AMP esculentin-1a, named esculentin-1a(1-21)NH2, [Esc(1-21): GIFSKLAGKKIKNLLISGLKG-NH2] has been found to have a potent activity against the Gram-negative bacterium Pseudomonas aeruginosa; a slightly weaker activity against Gram-positive bacteria and interesting immunomodulatory properties. With the aim to optimize the antimicrobial features of Esc(1-21) and to circumvent the limitations described above, two different approaches were followed: (i) substitutions by non-coded amino acids, i.e., α-aminoisobutyric acid or d-amino acids; and (ii) peptide conjugation to gold nanoparticles. In this mini-review, we summarized the structural and functional properties of the resulting Esc(1-21)-derived compounds. Overall, our data may assist researchers in the rational design and optimization of AMPs for the development of future drugs to fight the worldwide problem of antibiotic resistance
The frog skin-derived antimicrobial peptide esculentin-1a(1-21)nh2 promotes the migration of human hacat keratinocytes in an egf receptor-dependent manner: a novel promoter of human skin wound healing?
One of the many functions of skin is to protect the organism against a wide range of pathogens. Antimicrobial peptides (AMPs) produced by the skin epithelium provide an effective chemical shield against microbial pathogens. However, whereas antibacterial/antifungal activities of AMPs have been extensively characterized, much less is known regarding their wound healing-modulatory properties. By using an in vitro re-epithelialisation assay employing special cell-culture inserts, we detected that a derivative of the frog-skin AMP esculentin-1a, named esculentin-1a(1-21)NH2, significantly stimulates migration of immortalized human keratinocytes (HaCaT cells) over a wide range of peptide concentrations (0.025-4 μM), and this notably more efficiently than human cathelicidin (LL-37). This activity is preserved in primary human epidermal keratinocytes. By using appropriate inhibitors and an enzyme-linked immunosorbent assay we found that the peptide-induced cell migration involves activation of the epidermal growth factor receptor and STAT3 protein. These results suggest that esculentin-1a(1-21)NH2 now deserves to be tested in standard wound healing assays as a novel candidate promoter of skin re-epithelialisation. The established ability of esculentin-1a(1-21)NH2 to kill microbes without harming mammalian cells, namely its high anti-Pseudomonal activity, makes this AMP a particularly attractive candidate wound healing promoter, especially in the management of chronic, often Pseudomonas-infected, skin ulcers
Dynamic Modelling and Energy, Economic, and Environmental Analysis of a Greenhouse Supplied by Renewable Sources
Purification and characterization of recombinant human renin for X-ray crystallization studies
<p>Abstract</p> <p>Background</p> <p>The renin-angiotensin-aldosterone system (RAS) cascade is a major target for the clinical management of hypertension. Although inhibitors of various components of this cascade have been developed successfully, development of renin inhibitors has proven to be problematic. The development of these inhibitors has been hindered by poor bioavailability and complex synthesis. However, despite the challenges of designing renin inhibitors, the enzyme remains a promising target for the development of novel treatments for hypertension. X-ray crystallographic data could greatly assist the design and development of these inhibitors. Here we describe the purification and characterization of recombinant human renin for x-ray crystallization studies.</p> <p>Results</p> <p>A cDNA encoding the full length of native human preprorenin (406 amino acid residues) was introduced into the HEK-293 cell line. A clonal cell line expressing prorenin was generated and grown under serum free conditions in a hollow fiber bioreactor. Prorenin was constitutively secreted and purified directly from the conditioned medium. Concanavalin A chromatography effectively enriched and purified prorenin to 90% homogeneity in a single step. Prorenin was converted to active renin by trypsin digestion to remove the propeptide. Active renin was further purified using a cation exchange column followed by a gel filtration column. Biochemical characterization of the recombinant enzyme showed both binding and catalytic properties were essentially identical to previously reported activities for purified renin. Crystals were grown using this material in our X-ray structure studies, and high resolution diffraction was obtained.</p> <p>Conclusion</p> <p>This present work describes a simple and efficient method for the generation and purification of active human renin. The protein is highly pure and is suitable for supporting structural biology efforts.</p
Design of a cybernetic hand for perception and action
Strong motivation for developing new prosthetic hand devices is provided by the fact that low functionality and controllability—in addition to poor cosmetic appearance—are the most important reasons why amputees do not regularly use their prosthetic hands. This paper presents the design of the CyberHand, a cybernetic anthropomorphic hand intended to provide amputees with functional hand replacement. Its design was bio-inspired in terms of its modular architecture, its physical appearance, kinematics, sensorization, and actuation, and its multilevel control system. Its underactuated mechanisms allow separate control of each digit as well as thumb–finger opposition and, accordingly, can generate a multitude of grasps. Its sensory system was designed to provide proprioceptive information as well as to emulate fundamental functional properties of human tactile mechanoreceptors of specific importance for grasp-and-hold tasks. The CyberHand control system presumes just a few efferent and afferent channels and was divided in two main layers: a high-level control that interprets the user’s intention (grasp selection and required force level) and can provide pertinent sensory feedback and a low-level control responsible for actuating specific grasps and applying the desired total force by taking advantage of the intelligent mechanics. The grasps made available by the high-level controller include those fundamental for activities of daily living: cylindrical, spherical, tridigital (tripod), and lateral grasps. The modular and flexible design of the CyberHand makes it suitable for incremental development of sensorization, interfacing, and control strategies and, as such, it will be a useful tool not only for clinical research but also for addressing neuroscientific hypotheses regarding sensorimotor control
KDEON WK-11: A short antipseudomonal peptide with promising potential
The plight of antimicrobial resistance continues to limit the availability of antibiotic treatment effective in combating resistant bacterial infections. Despite efforts made to rectify this issue and minimise its effects on both patients and the wider community, progress in this area remains minimal. Here, we de-novo designed a peptide named KDEON WK-11, building on previous work establishing effective residues and structures active in distinguished antimicrobial peptides such as lactoferrin. We assessed its antimicrobial activity against an array of bacterial strains and identified its most potent effect, against Pseudomonas aeruginosa with an MIC value of 3.12 mu M, lower than its counterparts developed with similar residues and chain lengths. We then determined its anti-biofilm properties, potential mechanism of action and in vitro cytotoxicity. We identified that KDEON WK-11 had a broad range of antimicrobial activity and specific capabilities to fight Pseudomonas aeruginosa with low in vitro cytotoxicity and promising potential to express anti-lipopolysaccharide qualities, which could be exploited to expand its properties into an anti-sepsis agent
A Citizen Science Approach for Analyzing Social Media With Crowdsourcing
Social media have the potential to provide timely information about emergency situations and sudden events. However, finding relevant information among the millions of posts being added every day can be difficult, and in current approaches developing an automatic data analysis project requires time and technical skills. This work presents a new approach for the analysis of social media posts, based on configurable automatic classification combined with Citizen Science methodologies. The process is facilitated by a set of flexible, automatic and open-source data processing tools called the Citizen Science Solution Kit. The kit provides a comprehensive set of tools that can be used and personalized in different situations, particularly during natural emergencies, starting from images and text contained in the posts. The tools can be employed by citizen scientists for filtering, classifying, and geolocating the content with a human-in-the-loop approach to support the data analyst, including feedback and suggestions on how to configure the automated tools, and techniques to gather inputs from citizens. Using flooding scenario as a guiding example, this paper illustrates the structure and functioning of the different tools proposed to support citizens scientists in their projects, and a methodological approach to their use. The process is then validated by discussing three case studies based on the Albania earthquake of 2019, the Covid-19 pandemic, and the Thailand floods of 2021. The results suggest that a flexible approach to tools composition and configuration can support a timely setup of an analysis project by citizen scientists, especially in case of emergencies in unexpected locations.ISSN:2169-353
HDV can constrain HBV genetic evolution in hbsag: Implications for the identification of innovative pharmacological targets
Chronic HBV + HDV infection is associated with greater risk of liver fibrosis, earlier hepatic decompensation, and liver cirrhosis hepatocellular carcinoma compared to HBV mono-infection. However, to-date no direct anti-HDV drugs are available in clinical practice. Here, we identified conserved and variable regions in HBsAg and HDAg domains in HBV + HDV infection, a critical finding for the design of innovative therapeutic agents. The extent of amino-acid variability was measured by Shannon-Entropy (Sn) in HBsAg genotype-D sequences from 31 HBV + HDV infected and 62 HBV mono-infected patients (comparable for demographics and virological-parameters), and in 47 HDAg genotype-1 sequences. Positions with Sn = 0 were defined as conserved. The percentage of conserved HBsAg-positions was significantly higher in HBV + HDV infection than HBV mono-infection (p = 0.001). Results were confirmed after stratification for HBeAg-status and patients’ age. A Sn = 0 at specific positions in the C-terminus HBsAg were correlated with higher HDV-RNA, suggesting that conservation of these positions can preserve HDV-fitness. Conversely, HDAg was characterized by a lower percentage of conserved-residues than HBsAg (p < 0.001), indicating higher functional plasticity. Furthermore, specific HDAg-mutations were significantly correlated with higher HDV-RNA, suggesting a role in conferring HDV replicative-advantage. Among HDAg-domains, only the virus-assembly signal exhibited a high genetic conservation (75% of conserved-residues). In conclusion, HDV can constrain HBsAg genetic evolution to preserve its fitness. The identification of conserved regions in HDAg poses the basis for designing innovative targets against HDV-infection
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