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.     

Defuzzification as a special way of dealing with retranslation

The problem of defuzzification is examined in this paper from a broader perspective as a special way of dealing with the general problem of retranslation. The paper includes an overview of different formulations of the problem of defuzzification, as well as an overview of methods that have been suggested in the literature for dealing with the problem. Our own approach to defuzzification, which is described in the paper in more details, is based on relevant measures of uncertainty-based information. This paper is a companion to our recent paper that addresses the general problem of retranslation in computing with perceptions (Martin and Klir 2006).     

Lunar Habitat Micrometeoroid and Radiation Shielding: Options, Applications, and Assessments

Various shielding approaches to protect lunar habitats from micrometeoroid and radiation hazards present major trade-off considerations. Popular scenarios that envision covering modules with in situ regolith will necessitate means to excavate and move large amounts of material; will complicate evolutionary outpost growth; and may require long tunnels between connecting pressurized elements. Strategies that incorporate shielding materials into module structures or internal shelters add very substantial launch mass penalties. Utilization of water bladders can make efficient use of consumable/recyclable supplies, but may impose excess capacity deliveries at early development stages. This paper addresses these different shielding approaches from a top-level application perspective, highlighting pros and cons of each. Examples draw upon research and design investigations undertaken by the Sasakawa International Center for Space Architecture in support of separate National Aeronautics and Space Administration (NASA) contracts awarded to teams headed by Boeing and ILC-Dover for a “Minimum Functionality Habitation Systems Concept Study.” Comprehensive team study results were presented to NASA in February 2009, and have been released as public information.     

Longitudinal stability of temperamental exuberance and social–emotional outcomes in early childhood.

The goals of the current study were to investigate the stability of temperamental exuberance across infancy and toddlerhood and to examine the associations between exuberance and social–emotional outcomes in early childhood. The sample consisted of 291 4-month-olds followed at 9, 24, and 36 months and again at 5 years of age. Behavioral measures of exuberance were collected at 9, 24, and 36 months. At 36 months, frontal electroencephalogram (EEG) asymmetry was assessed. At 5 years, maternal reports of temperament and behavior problems were collected, as were observational measures of social behavior during an interaction with an unfamiliar peer in the laboratory. Latent profile analysis revealed a high, stable exuberance profile that was associated with greater ratings of 5-year externalizing behavior and surgency, as well as observed disruptive behavior and social competence with unfamiliar peers. These associations were particularly true for children who displayed left frontal EEG asymmetry. Multiple factors supported an approach bias for exuberant temperament but did not differentiate between adaptive and maladaptive social–emotional outcomes at 5 years of age. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

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.     

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.     

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.