Recent Biophysical Issues About the Preparation of Solute-Filled Lipid Vesicles

Here we report some recent biophysical issues on the preparation of solute-filled lipid vesicles and their relevance to the construction of “synthetic cells.” First, we introduce the “semi-synthetic minimal cells” as the liposome-based cell-like systems, which contain a minimal number of biomolecules required to display simple and complex biological functions. Next, we focus on recent aspects related to the construction of synthetic cells. Emphasis is given to the interplay between the methods of synthetic cell preparation and the physics of solute encapsulation. We briefly introduce the notion of structural and compositional “diversity” in synthetic cell populations.     

Preparation and characterization of chitosan‐based dispersions

Complexation of chitosan in aqueous solutions by low molecular weight electrolytes is one of the simplest methods for the preparation of aqueous chitosan dispersions. In this work, the influence of storage time, sulfate concentration, method of preparation and surfactant content on some properties of the resultant chitosan dispersions (turbidity, viscosity and zeta potential) was analyzed. Turbidimetry was adequate to monitor the formation of particles, while viscometry was suitable to monitor changes in the dispersing phase. An analysis of the properties of these systems, mainly in terms of particle–particle and macromolecule–macromolecule interactions was carried out. Copyright © 2004 Society of Chemical Industry     

Diagnosing Czech Patients with Inherited Platelet Disorders

A single-center study was conducted on 120 patients with inherited disorders of primary hemostasis followed at our hematological center. These patients presented a variety of bleeding symptoms; however, they had no definitive diagnosis. Establishing a diagnosis has consequences for the investigation of probands in families and for treatment management; therefore, we aimed to improve the diagnosis rate in these patients by implementing advanced diagnostic methods. According to the accepted international guidelines at the time of study, we investigated platelet morphology, platelet function assay, light-transmission aggregometry, and flow cytometry. Using only these methods, we were unable to make a definitive diagnosis for most of our patients. However, next-generation sequencing (NGS), which was applied in 31 patients, allowed us to establish definitive diagnoses in six cases (variants in ANKRD26, ITGA2B, and F8) and helped us to identify suspected variants (NBEAL2, F2, BLOC1S6, AP3D1, GP1BB, ANO6, CD36, and ITGB3) and new suspected variants (GFI1B, FGA, GP1BA, and ITGA2B) in 11 patients. The role of NGS in patients with suspicious bleeding symptoms is growing and it changes the diagnostic algorithm. The greatest disadvantage of NGS, aside from the cost, is the occurrence of gene variants of uncertain significance.     

Aminopeptidase N Inhibitors as Pointers for Overcoming Antitumor Treatment Resistance

Aminopeptidase N (APN), also known as CD13 antigen or membrane alanyl aminopeptidase, belongs to the M1 family of the MA clan of zinc metallopeptidases. In cancer cells, the inhibition of aminopeptidases including APN causes the phenomenon termed the amino acid deprivation response (AADR), a stress response characterized by the upregulation of amino acid transporters and synthetic enzymes and activation of stress-related pathways such as nuclear factor kB (NFkB) and other pro-apoptotic regulators, which leads to cancer cell death by apoptosis. Recently, APN inhibition has been shown to augment DR4-induced tumor cell death and thus overcome resistance to cancer treatment with DR4-ligand TRAIL, which is available as a recombinant soluble form dulanermin. This implies that APN inhibitors could serve as potential weapons for overcoming cancer treatment resistance. In this study, a series of basically substituted acetamidophenones and the semicarbazones and thiosemicarbazones derived from them were prepared, for which APN inhibitory activity was determined. In addition, a selective anti-proliferative activity against cancer cells expressing APN was demonstrated. Our semicarbazones and thiosemicarbazones are the first compounds of these structural types of Schiff bases that were reported to inhibit not only a zinc-dependent aminopeptidase of the M1 family but also a metalloenzyme.     

Active application of primer acid on acid‐treated enamel: Influence on the bond effectiveness of self‐etch adhesives systems

Aim: Evaluate the composite‐to‐enamel bond after passive or active application of self‐etching primer systems on polished or pre‐etched enamel with phosphoric acid. Materials and Methods: Two self‐etch adhesives systems (SEAS) were used: Clearfil SE Bond and Easy Bond. Third human molars were divided into 8 groups (N = 10). The crown of each tooth was sectioned into halves and the mesial/distal surfaces were used. The adhesives were actively or passively applied on enamel with or without prior phosphoric‐acid etching. Resin composite cylinders were built after adhesive application. After stored in relative humidity for 24 hr/37°C the specimens were subjected to microshear test in universal testing a machine at a crosshead speed of 0.5 mm/minute. The results were analyzed with three‐way ANOVA and the Tukey test. The enamel‐etching pattern was evaluated under SEM. Results: The 2‐step SEAS system presented significantly higher adhesive bond strength means (47.37 MPa) than the 1‐step (36.87 MPa). A poor enamel‐ etching pattern was observed in active mode showing irregular and short resin tags, however there was not compromised the bond strength. Conclusion: Active or passive application produced similar values of bond strength to enamel regardless of enamel pretreatment and type of SEAS.     

Survey and comparison of message authentication solutions on wireless sensor networks

Security is an important concern in any modern network. This also applies to Wireless Sensor Networks (WSNs), especially those used in applications that monitor sensitive information (e.g., health care applications). However, the highly constrained nature of sensors imposes a difficult challenge: their reduced availability of memory, processing power and energy hinders the deployment of many modern cryptographic algorithms considered secure. For this reason, the choice of the most memory-, processing- and energy-efficient security solutions is of vital importance in WSNs. To date, a number of extensive analyses comparing different encryption algorithms and key management schemes have been developed, while very little attention has been given to message authentication solutions. In this paper, aiming to close this gap, we identify cipher-based Message Authentication Codes (MACs) and Authenticated Encryption with Associated Data (AEAD) schemes suitable for WSNs and then evaluate their features and performance on a real platform (TelosB). As a result of this analysis, we identify the recommended choices depending on the characteristics of the target network and available hardware.     

ENERGY LOSS AND ENERGY STRAGGLING OF LIGHT IONS IN FULLERITE

To determinate the stopping cross section in fullerite a feasible approach, taking into account the high radiation sensitivity and mechanical fragility of fullerite films, should be employed. In this work, the stopping cross sections of ¹H, ³H, ⁴He and ⁷Li ions for several selected energies were measured by Rutherford backscattering, neutron depth profiling and alpha spectroscopy using sandwich structures of samples composed of fullerite deposited on a firm substrate (Si, steel) with an intermediate marker (Au, N, Li, B, Pu). In addition, ion transmission through a thin C film supporting a fullerite layer was also utilized. The measured stopping cross sections follow the theoretical predictions calculated for carbon, but are systematically (10-35%) higher than the theoretical ones (with the exception of 5 — 5.5 MeV ⁴He). The observed deviation of the experimental data can partly be explained by the chemical state effects in fullerite, which accounts for about 20-50% of the difference. The measured energy straggling exceeds Bohr's value by a factor of about 2 for alpha spectroscopy and ion transmission, and 2.5 or 9.5 for Rutherford backscattering and neutron depth profiling, respectively. The discrepancy can be explained by a thickness variation, such as surface roughness of the fullerite films.     

Comparison of Consumer Attitudes Between Cyprus and Latvia: An Evaluation of Effect of Setting on Consumer Preferences in the Water Industry

Models approaching consumer expectations of their water supplier from a risk perspective suggest that consumers primarily and overwhelmingly want safe drinking water supply. In this study consumer preferences in the water sector are investigated in two contrasting case studies: Cyprus, where there have been significant quantity and continuity of supply issues, and Riga, where there have been water quality issues. While water quality is undoubtedly the main priority of water consumers in Riga, in Cyprus consumers indicate that they prioritise a more reliable service even though many are sufficiently dissatisfied with water quality that they do not drink the tap water. The analysis of consumer attitudes in the two case studies suggests that when water supply is unreliable, reliability takes precedence; once it is reliable quality issues come to the fore.     

Aerobic degradation of 2,4-dinitrotoluene by individual bacterial strains and defined mixed population in submerged cultures

The degradation efficiencies of isomeric mononitrotoluenes (2- and 4-NTs) and dinitrotoluenes (2,4-DNT and 2,6-DNT) by either individual bacterial strains (Bacillus cereus NDT4, Pseudomonas putida NDT1, Pseudomonas fluorescens NDT2, and Achromobacter sp. NDT3) or their mixture were compared in submerged batch cultivations. The mixed culture degraded 2,4-DNT nearly 50 times faster than any of the individual strains. The mixed culture also demonstrated significantly shorter lag periods in 2,4-DNT degradation, a lack of nitrite or organic intermediates accumulation in the liquid medium and the ability to degrade a broader spectrum of nitrotoluenes over a wider concentration range. The presence of both readily degradable 2-NT (or 4-NT) and poorly degradable 2,6-DNT in the medium negatively affected 2,4-DNT biodegradation. However, the mixed bacterial culture still effectively degraded 2,4-DNT with only slightly lower rates under these unfavorable conditions, thus showing potential for the remediation of 2,4-DNT contaminated sites.     

Detection of Campylobacter jejuni in naturally contaminated chicken skin by melting peak analysis of amplicons in real-time PCR

Contamination of poultry by Campylobacter spp. is a significant source of human diarrheal diseases. Traditional methods currently used to detect Campylobacter in foods are time-consuming and labor-intensive. In this study, primers designed for the Campylobacter jejuni cadF gene sequence were used in a SYBR Green I real-time PCR assay as an alternative to a conventional bacteriological method for the rapid detection of C. jejuni from poultry. Twelve portions of chicken purchased from two local grocery stores and 39 portions obtained from a commercial processing plant were examined. Samples of the skin were enriched in Bolton broth at 37 degrees C for 3 h and then at 42 degrees C for 9, 21, or 45 h under microaerobic conditions. DNA was extracted from 1-ml aliquots of the enrichment cultures using 1% Triton X-100. The DNA was used as the template in a real-time polymerase chain reaction (PCR) assay. After 24 h of enrichment, C. jejuni was isolated from 13 samples and all of the positive cultures were also detected by the real-time PCR procedure. C. jejuni was detected by both methods from samples artificially contaminated with 1 or 10 CFU of C. jejuni per 10 g, after 24 h of enrichment. The real-time PCR method was found to be sensitive and specific. It significantly reduced the time required for the detection of C. jejuni in poultry following enrichment of samples.