RESUMEN
OBJECTIVES: Quantification of plasma hepatitis D virus (HDV) RNA is the essential tool for patient management under antiviral therapy. The aim of this European multicenter study was to improve the comparability of quantitative results reported by different laboratories using the CE/IVD-labeled RoboGene HDV RNA Quantification Kit 2.0 (Roboscreen GmbH) with different manual or automated nucleic acid extraction protocols/platforms and amplification/detection devices. METHODS: For harmonization of HDV RNA concentrations obtained by different protocols, correction factors (CF) were determined using the 1st WHO International Standard for HDV RNA. The limit of detection (LOD) and accuracy were determined for each protocol by using reference material. Furthermore, clinical samples were analyzed and results compared. RESULTS: The CF ranged from 20 to 1,870 depending on the protocol used. The LOD was found between 4 and 450 IU/ml. When accuracy was tested, external quality control (EQC) samples containing low HDV RNA concentrations were not detected by those protocols with higher LODs. For EQC samples, the maximum standard deviation of HDV RNA concentrations was found to be 0.53 log10 IU/ml, for clinical samples 0.87 log10 IU/mL. CONCLUSION: To ensure reliability in quantification of HDV RNA, any modification of the extraction and amplification/detection protocol validated by the manufacturer requires revalidation. With the 1st WHO International Standard for HDV RNA, the CF could easily be calculated leading to harmonization of quantitative results. This warrants both accurate monitoring of response to existing anti-HDV treatment and comparability of study results investigating novel anti-HDV drugs.
Asunto(s)
Hepatitis D , Preparaciones Farmacéuticas , Hepatitis D/diagnóstico , Hepatitis D/tratamiento farmacológico , Virus de la Hepatitis Delta/genética , Humanos , ARN Viral , Reproducibilidad de los Resultados , Carga ViralRESUMEN
Multidrug resistant (MDR) bacteria have adapted to most clinical antibiotics and are a growing threat to human health. One promising type of candidates for the everlasting demand of new antibiotic compounds constitute antimicrobial peptides (AMPs). These peptides act against different types of microbes by permeabilizing pathogen cell membranes, whereas being harmless to mammalian cells. Contrarily, another class of membrane-active peptides, namely cell-penetrating peptides (CPPs), is known to translocate in eukaryotic cells without substantially affecting the cell membrane. Since CPPs and AMPs share several physicochemical characteristics, we hypothesized if we can rationally direct the activity of a CPP towards antimicrobial activity. Herein, we describe the screening of a synthetic library, based on the CPP sC18, including structure-based design to identify the active residues within a CPP sequence and to discover novel AMPs with high activity. Peptides with increased hydrophobicity were tested against various bacterial strains, and hits were further optimized leading to four generations of peptides, with the last also comprising fluorinated amino acid building blocks. Interestingly, beside strong antibacterial activities, we also detected activity in cancer cells, while non-cancerous cells remained unharmed. The results highlight our new candidates, particularly those from generation 4, as a valuable and promising source for the development of future therapeutics with antibacterial activity and beyond.
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Antibacterianos/farmacología , Péptidos Catiónicos Antimicrobianos/química , Antineoplásicos/farmacología , Bacterias/efectos de los fármacos , Membrana Celular/efectos de los fármacos , Péptidos de Penetración Celular/química , Péptidos Catiónicos Antimicrobianos/síntesis química , Péptidos Catiónicos Antimicrobianos/farmacología , Bacillus subtilis/efectos de los fármacos , Bacillus subtilis/ultraestructura , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Péptidos de Penetración Celular/síntesis química , Péptidos de Penetración Celular/farmacología , Dicroismo Circular , Corynebacterium glutamicum/efectos de los fármacos , Corynebacterium glutamicum/ultraestructura , Halogenación , Hemólisis/efectos de los fármacos , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Concentración 50 Inhibidora , Pruebas de Sensibilidad Microbiana , Micrococcus luteus/efectos de los fármacos , Microscopía Electrónica de Rastreo , Pseudomonas fluorescens/efectos de los fármacos , Pseudomonas fluorescens/ultraestructuraRESUMEN
In this study, the efficacy of electron beam irradiation versus chemical coupling for yielding polyethersulfone (PES) membranes with antibacterial properties was investigated. For the surface coating, a recently discovered lead compound, IL-KKA, comprising a short peptide sequence functionalized with imidazolium groups, was used. For better integration within the membrane, several novel variants of IL-KKA were generated. Membrane immobilization was achieved using different doses of electron beam irradiation and NHS/EDC chemical coupling. Physicochemical characterization of the coated membranes was performed by water contact angle measurements, X-ray photoelectron spectroscopy, and scanning electron microscopy. Our results show that electron beam irradiation is as effective and gentle as chemical coupling using the NHS/EDC method. Moreover, it was demonstrated that the obtained membranes exhibit promising antibacterial activity against B. subtilis. In summary, the technique presented herein might be promising as a template for developing future anti-biofilm devices.
RESUMEN
Antimicrobial peptides (AMPs) are an interesting class of antibiotics characterized by their unique antibiotic activity and lower propensity for developing resistance compared to common antibiotics. They belong to the class of membrane-active peptides and usually act selectively against bacteria, fungi and protozoans. AMPs, but also peptide conjugates containing AMPs, have come more and more into the focus of research during the last few years. Within this article, recent work on AMP conjugates is reviewed. Different aspects will be highlighted as a combination of AMPs with antibiotics or organometallic compounds aiming to increase antibacterial activity or target selectivity, conjugation with photosensitizers for improving photodynamic therapy (PDT) or the attachment to particles, to name only a few. Owing to the enormous resonance of antimicrobial conjugates in the literature so far, this research topic seems to be very attractive to different scientific fields, like medicine, biology, biochemistry or chemistry.