High Precision Human Skin Temperature Fluctuations Measuring Instrument.

This paper describes the experimental results of testing a prototype of a high precision human skin rapid temperature fluctuations measuring instrument. Based on the author's work, an original circuit solution on a miniature semiconductor diode sensor has been designed. The proposed circuitry provides operation in the full voltage range with automatic setting and holding the operating point, as well as the necessary slope of the conversion coefficient (up to 2300 mV/°C), which makes it possible to register fast temperature oscillations from the surface of the human body and other biological objects. Simulation results in the Microcap 12 software and laboratory tests have confirmed all declared design specifications: temperature resolution of 0.01 °C, transducer thermal time constant of 0.05 s. An original thermostat and an experimental setup for the simultaneous registration of the electrocardiogram, pulse wave signals from the Biopac polygraph MP36 and a signal of temperature oscillations from the prototype thermometer have been designed for further investigations. The preliminary test results indicates that using the designed measuring instrument gives a possibility to provide an in-depth study of the relationship between micro- and macro-blood circulations manifested in skin temperature fluctuations.

Study of the Fabrication Technology of Hybrid Microfluidic Biochips for Label-Free Detection of Proteins.

A study of the peculiarities and a comparative analysis of the technologies used for the fabrication of elements of novel hybrid microfluidic biochips for express biomedical analysis have been carried out. The biochips were designed with an incorporated microfluidic system, which enabled an accumulation of the target compounds in a biological fluid to be achieved, thus increasing the biochip system's sensitivity and even implementing a label-free design of the detection unit. The multilevel process of manufacturing a microfluidic system of a given topology for label-free fluorometric detection of protein structures is presented. The technological process included the chemical modification of the working surface of glass substrates by silanization using (3-aminopropyl) trimethoxysilane (APTMS), formation of the microchannels, for which SU-8 technologies and a last generation dry film photoresist were studied and compared. The solid-state phosphor layers were deposited using three methods: drop application; airbrushing; and mechanical spraying onto the adhesive surface. The processes of sealing the system, installing input ports, and packaging using micro-assembly technologies are described. The technological process has been optimized and the biochip was implemented and tested. The presented system can be used to design novel high-performance diagnostic tools that implement the function of express detection of protein markers of diseases and create low-power multimodal, highly intelligent portable analytical decision-making systems in medicine.

Comparison of AlN vs. SIO2/LiNbO3 Membranes as Sensitive Elements for the SAW-Based Acceleration Measurement: Overcoming the Anisotropy Effects.

We propose the use of aluminum nitride (AlN) membranes acting as sensitive elements for the surface acoustic wave (SAW)-based acceleration measurement. The proposed solution is compared against existing prototypes based on the use of quartz (SiO2)/lithium niobate (LiNbO3) membranes that are characterized by extensive anisotropic properties. Using COMSOL Multiphysics 5.4 computer simulations we show explicitly that sensitive elements based on less anisotropic AlN membranes overcome both the low sensitivity limitations of SiO2 and low temperature stability of LiNbO3. Moreover, AlN membranes exhibit nearly double the robustness against irreversible mechanical deformations when compared against SiO2, which in turn allows for further 1.5-fold sensitivity enhancement over LiNbO3 based sensors. Taking into account their acceptable frequency characteristics, we thus believe that the AlN membranes are a good candidate forsensitive elements especially for high acceleration measurements.

Fast and simple tool for the quantification of biofilm-embedded cells sub-populations from fluorescent microscopic images.

Fluorescent staining is a common tool for both quantitative and qualitative assessment of pro- and eukaryotic cells sub-population fractions by using microscopy and flow cytometry. However, direct cell counting by flow cytometry is often limited, for example when working with cells rigidly adhered either to each other or to external surfaces like bacterial biofilms or adherent cell lines and tissue samples. An alternative approach is provided by using fluorescent microscopy and confocal laser scanning microscopy (CLSM), which enables the evaluation of fractions of cells subpopulations in a given sample. For the quantitative assessment of cell fractions in microphotographs, we suggest a simple two-step algorithm that combines single cells selection and the statistical analysis. To facilitate the first step, we suggest a simple procedure that supports finding the balance between the detection threshold and the typical size of single cells based on objective cell size distribution analysis. Based on a series of experimental measurements performed on bacterial and eukaryotic cells under various conditions, we show explicitly that the suggested approach effectively accounts for the fractions of different cell sub-populations (like the live/dead staining in our samples) in all studied cases that are in good agreement with manual cell counting on microphotographs and flow cytometry data. This algorithm is implemented as a simple software tool that includes an intuitive and user-friendly graphical interface for the initial adjustment of algorithm parameters to the microphotographs analysis as well as for the sequential analysis of homogeneous series of similar microscopic images without further user intervention. The software tool entitled BioFilmAnalyzer is freely available online at https://bitbucket.org/rogex/biofilmanalyzer/downloads/.

Corrigendum: Superstatistical model of bacterial DNA architecture.

This corrects the article DOI: 10.1038/srep43034.

Superstatistical model of bacterial DNA architecture.

Understanding the physical principles that govern the complex DNA structural organization as well as its mechanical and thermodynamical properties is essential for the advancement in both life sciences and genetic engineering. Recently we have discovered that the complex DNA organization is explicitly reflected in the arrangement of nucleotides depicted by the universal power law tailed internucleotide interval distribution that is valid for complete genomes of various prokaryotic and eukaryotic organisms. Here we suggest a superstatistical model that represents a long DNA molecule by a series of consecutive ~150 bp DNA segments with the alternation of the local nucleotide composition between segments exhibiting long-range correlations. We show that the superstatistical model and the corresponding DNA generation algorithm explicitly reproduce the laws governing the empirical nucleotide arrangement properties of the DNA sequences for various global GC contents and optimal living temperatures. Finally, we discuss the relevance of our model in terms of the DNA mechanical properties. As an outlook, we focus on finding the DNA sequences that encode a given protein while simultaneously reproducing the nucleotide arrangement laws observed from empirical genomes, that may be of interest in the optimization of genetic engineering of long DNA molecules.

Statistical prediction of protein structural, localization and functional properties by the analysis of its fragment mass distributions after proteolytic cleavage.

Structural, localization and functional properties of unknown proteins are often being predicted from their primary polypeptide chains using sequence alignment with already characterized proteins and consequent molecular modeling. Here we suggest an approach to predict various structural and structure-associated properties of proteins directly from the mass distributions of their proteolytic cleavage fragments. For amino-acid-specific cleavages, the distributions of fragment masses are determined by the distributions of inter-amino-acid intervals in the protein, that in turn apparently reflect its structural and structure-related features. Large-scale computer simulations revealed that for transmembrane proteins, either α-helical or ß -barrel secondary structure could be predicted with about 90% accuracy after thermolysin cleavage. Moreover, 3/4 intrinsically disordered proteins could be correctly distinguished from proteins with fixed three-dimensional structure belonging to all four SCOP structural classes by combining 3-4 different cleavages. Additionally, in some cases the protein cellular localization (cytosolic or membrane-associated) and its host organism (Firmicute or Proteobacteria) could be predicted with around 80% accuracy. In contrast to cytosolic proteins, for membrane-associated proteins exhibiting specific structural conformations, their monotopic or transmembrane localization and functional group (ATP-binding, transporters, sensors and so on) could be also predicted with high accuracy and particular robustness against missing cleavages.