Cloning and Characterization of a Thermostable Endolysin of Bacteriophage TP-84 as a Potential Disinfectant and Biofilm-Removing Biological Agent

The obligatory step in the life cycle of a lytic bacteriophage is the release of its progeny particles from infected bacterial cells. The main barrier to overcome is the cell wall, composed of crosslinked peptidoglycan, which counteracts the pressure prevailing in the cytoplasm and protects the cell against osmotic lysis and mechanical damage. Bacteriophages have developed two strategies leading to the release of progeny particles: the inhibition of peptidoglycan synthesis and enzymatic cleavage by a bacteriophage-coded endolysin. In this study, we cloned and investigated the TP84_28 endolysin of the bacteriophage TP-84, which infects thermophilic Geobacillus stearothermophilus, determined the enzymatic characteristics, and initially evaluated the endolysin application as a non-invasive agent for disinfecting surfaces, including those exposed to high temperatures. Both the native and recombinant TP84_28 endolysins, obtained through the Escherichia coli T7-lac expression system, are highly thermostable and retain trace activity after incubation at 100 °C for 30 min. The proteins exhibit strong bacterial wall digestion activity up to 77.6 °C, decreasing to marginal activity at ambient temperatures. We assayed the lysis of various types of bacteria using TP84_28 endolysins: Gram-positive, Gram-negative, encapsulated, and pathogenic. Significant lytic activity was observed on the thermophilic and mesophilic Gram-positive bacteria and, to a lesser extent, on the thermophilic and mesophilic Gram-negative bacteria. The thermostable TP84_28 endolysin seems to be a promising mild agent for disinfecting surfaces exposed to high temperatures.     

In Vitro Cytotoxicity,Colonisation by Fibroblasts and Antimicrobial Properties of Surgical Meshes Coated with Bacterial Cellulose

Hernia repairs are the most common abdominal wall elective procedures performed by general surgeons. Hernia-related postoperative infective complications occur with 10% frequency. To counteract the risk of infection emergence, the development of effective, biocompatible and antimicrobial mesh adjuvants is required. Therefore, the aim of our in vitro investigation was to evaluate the suitability of bacterial cellulose (BC) polymer coupled with gentamicin (GM) antibiotic as an absorbent layer of surgical mesh. Our research included the assessment of GM-BC-modified meshes’ cytotoxicity against fibroblasts ATCC CCL-1 and a 60-day duration cell colonisation measurement. The obtained results showed no cytotoxic effect of modified meshes. The quantified fibroblast cells levels resembled a bimodal distribution depending on the time of culturing and the type of mesh applied. The measured GM minimal inhibitory concentration was 0.47 µg/mL. Results obtained in the modified disc-diffusion method showed that GM-BC-modified meshes inhibited bacterial growth more effectively than non-coated meshes. The results of our study indicate that BC-modified hernia meshes, fortified with appropriate antimicrobial, may be applied as effective implants in hernia surgery, preventing risk of infection occurrence and providing a high level of biocompatibility with regard to fibroblast cells.     

Design of reconfigurable machining lines: A novel comprehensive optimisation method

We present a novel comprehensive optimization model for designing reconfigurable machining lines. Due to the proposed fine mathematical modelling, it is possible to optimize simultaneously the whole set of machines and machining modules as well as their cutting parameters, their configuration that will be used for processing of each part and part position at each machine. The experimental results show that the proposed optimization approach substantially outperforms the existing heuristic design method and therefore it can be used by the designers in order to reduce the total system cost and improve the efficiency of reconfigurable machining lines.     

Phylogenetic-Derived Insights into the Evolution of Sialylation in Eukaryotes: Comprehensive Analysis of Vertebrate β-Galactoside α2,3/6-Sialyltransferases (ST3Gal and ST6Gal)

Cell surface of eukaryotic cells is covered with a wide variety of sialylated molecules involved in diverse biological processes and taking part in cell–cell interactions. Although the physiological relevance of these sialylated glycoconjugates in vertebrates begins to be deciphered, the origin and evolution of the genetic machinery implicated in their biosynthetic pathway are poorly understood. Among the variety of actors involved in the sialylation machinery, sialyltransferases are key enzymes for the biosynthesis of sialylated molecules. This review focus on β-galactoside α2,3/6-sialyltransferases belonging to the ST3Gal and ST6Gal families. We propose here an outline of the evolutionary history of these two major ST families. Comparative genomics, molecular phylogeny and structural bioinformatics provided insights into the functional innovations in sialic acid metabolism and enabled to explore how ST-gene function evolved in vertebrates.     

Factors Influencing Rapid Detection of Bioaerosols by Surface Plasmon Resonance-Based Technique

In bioaerosol monitoring applications, technologies allowing rapid and precise detection of airborne pathogens are highly demanded. One of such technologies, based on the immunoreaction-operating principle in nearly real-time mode without any specific labeling, is known as surface plasmon resonance (SPR). In previous studies, we have shown applicability of the SPR technology for rapid and selective detection of viral and bacterial aerosols where successful combination of the SPR machine with our earlier produced personal bioaerosol sampler opened new prospects in development of portable bioaerosol monitors. The current study is a logical continuation of our previous research dedicated to the technology development for rapid bioaerosol detection. Here, we focus on one of the main factors possibly influencing the SPR-based bioaerosol monitoring; the SPR performance on target bioaerosol detection was evaluated at conditions of substantial air contamination with different nontargeted microorganisms, commonly presented in the air. Besides, different compositions of sampling liquids were tested in regards to the SPR results interference. Our findings clearly verified high specificity of the technology even in cases of highly contaminated air environments with aerosols of biological and mineral origins. It was found that both nontargeted bioaerosols and nanosized aerosols of mineral background do not have significant influence on the specific SPR detection of targeted bioaerosols.

Copyright 2014 American Association for Aerosol Research     

Using light to control the interactions between self-rotating assemblies of active gels

Polymer gels undergoing the oscillating Belousov-Zhabotinsky (BZ) reaction exhibit an autonomous, periodic swelling and deswelling, where the mechanical oscillations are driven by the chemical reaction within the polymer network. Using computer simulations, we show that these BZ gels can undergo a form of auto-chemotaxis, enabling the gels to spontaneously move in response to self-generated chemical gradients. Focusing on four millimeter-sized pieces of these BZ gels, we show that the pieces can organize into self-rotating clusters that resemble a moving pinwheel or gear. By analyzing the factors that promote the formation of a single self-rotating cluster, we attempt to design systems of multiple, interacting gears. We show that light, which suppresses the oscillations of the gels, can be harnessed to promote the formation of two self-rotating clusters. These studies point to a novel form of photo-chemo-mechanical transduction, where light is utilized to control the conversion of chemical and mechanical energy in the system. Moreover, the interaction between the BZ gel gears reveals a new form of entrainment between these moving units. Namely, their coordinated motion is achieved through chemical coupling or communication, rather than a mechanical coupling. These findings can lead to the formation of chemically “communicating” devices that can be programmed to perform autonomous work through the use of light.     

A process for the complete fractionation of baker's yeast

A process for the preparation of yeast-derived food additives was developed. The four products obtained, yeast protein concentrate (YPC), cell-wall protein (CWP), semi-pure glucomannan (SPG) and yeast extract, compared well with similar products, present in the current market, with respect to functional properties. YPC and CWP exhibited improved water-holding and oil-binding properties over those of soy-protein isolate (SPI). The emulsifying capacity was very close to that observed in commercial samples. A one-step alkaline extraction enhanced the functional properties of primary yeast glycan to match traditional food hydrocolloid sources. SPG showed oil-binding properties significantly greater than commercial glucomannan.     

From miRNA Target Gene Network to miRNA Function: miR-375 Might Regulate Apoptosis and Actin Dynamics in the Heart Muscle via Rho-GTPases-Dependent Pathways

MicroRNAs (miRNAs) are short, single-stranded, non-coding ribonucleic acid (RNA) molecules, which are involved in the regulation of main biological processes, such as apoptosis or cell proliferation and differentiation, through sequence-specific interaction with target mRNAs. In this study, we propose a workflow for predicting miRNAs function by analyzing the structure of the network of their target genes. This workflow was applied to study the functional role of miR-375 in the heart muscle (myocardium), since this miRNA was previously shown to be associated with heart diseases, and data on its function in the myocardium are mostly unclear. We identified PIK3CA, RHOA, MAPK3, PAFAH1B1, CTNNB1, MYC, PRKCA, ERBB2, and CDC42 as key genes in the miR-375 regulated network and predicted the possible function of miR-375 in the heart muscle, consisting mainly in the regulation of the Rho-GTPases-dependent signaling pathways. We implemented our algorithm for miRNA function prediction into a Python module, which is available at GitHub.     

eDNA Inactivation and Biofilm Inhibition by the PolymericBiocide Polyhexamethylene Guanidine Hydrochloride (PHMG-Cl)

The choice of effective biocides used for routine hospital practice should consider the role of disinfectants in the maintenance and development of local resistome and how they might affect antibiotic resistance gene transfer within the hospital microbial population. Currently, there is little understanding of how different biocides contribute to eDNA release that may contribute to gene transfer and subsequent environmental retention. Here, we investigated how different biocides affect the release of eDNA from mature biofilms of two opportunistic model strains Pseudomonas aeruginosa ATCC 27853 (PA) and Staphylococcus aureus ATCC 25923 (SA) and contribute to the hospital resistome in the form of surface and water contaminants and dust particles. The effect of four groups of biocides, alcohols, hydrogen peroxide, quaternary ammonium compounds, and the polymeric biocide polyhexamethylene guanidine hydrochloride (PHMG-Cl), was evaluated using PA and SA biofilms. Most biocides, except for PHMG-Cl and 70% ethanol, caused substantial eDNA release, and PHMG-Cl was found to block biofilm development when used at concentrations of 0.5% and 0.1%. This might be associated with the formation of DNA–PHMG-Cl complexes as PHMG-Cl is predicted to bind to AT base pairs by molecular docking assays. PHMG-Cl was found to bind high-molecular DNA and plasmid DNA and continued to inactivate DNA on surfaces even after 4 weeks. PHMG-Cl also effectively inactivated biofilm-associated antibiotic resistance gene eDNA released by a pan-drug-resistant Klebsiella strain, which demonstrates the potential of a polymeric biocide as a new surface-active agent to combat the spread of antibiotic resistance in hospital settings.