First vertebrate BRICHOS antimicrobial peptides: ß-hairpin host defense peptides in limbless amphibia lung resemble those of marine worms.

Innate immunity of invertebrates offers potent antimicrobial peptides (AMPs) against drug-resistant infections. To identify new worm ß-hairpin AMPs, we explored the sequence diversity of proteins with a BRICHOS domain, which comprises worm AMP precursors. Strikingly, we discovered new BRICHOS AMPs not in worms, but in caecilians, the least studied clade of vertebrates. Two precursor proteins from Microcaecilia unicolor and Rhinatrema bivittatum resemble SP-C lung surfactants and bear worm AMP-like peptides at C-termini. The analysis of M. unicolor tissue transcriptomes shows that the AMP precursor is highly expressed in the lung along with regular SP-C, suggesting a different, protective function. The peptides form right-twisted ß-hairpins, change conformation upon lipid binding, and rapidly disrupt bacterial membranes. Both peptides exhibit broad-spectrum activity against multidrug-resistant ESKAPE pathogens with 1-4 µM MICs and remarkably low toxicity, giving 40-70-fold selectivity towards bacteria. These BRICHOS AMPs, previously unseen in vertebrates, reveal a novel lung innate immunity mechanism and offer a promising antibiotics template.

A Simple Contactless High-Frequency Electromagnetic Sensor: Proof of Concept.

We report on the development of a very simple and inexpensive sensor device based on an inductance coil connected to a high-frequency electric field generator. The working principle of this sensor is as follows: liquid sample in a plastic tube is placed inside the inductance coil as its core and this core changes the properties of high-frequency electric current passing through the coil; these changes depend on sample conductivity, dielectric constant, magnetic properties, and capacitance. The electric signal registered after the coil represents a kind of a spectrum that can be effectively treated with chemometric tools. Our studies have shown that such a sensor can be used to distinguish between substances with different physical and chemical properties; different concentrations of ions in aqueous solutions with a linear response in a broad concentration range from 10-3 M to 10-1 M; can quantify the fat content in milk and cream samples with about 2% accuracy; and can distinguish between bacterial cultures and cell line cultures. The measurements are very fast and contactless, and the results show good promise for developing a number of applications of this sensor system.

Rapid Evaluation of Integral Quality and Safety of Surface and Waste Waters by a Multisensor System (Electronic Tongue).

The paper describes a wide-range practical application of the potentiometric multisensor system (MS) (1) for integral safety evaluation of a variety of natural waters at multiple locations, under various climatic conditions and anthropogenic stress and (2) for close to real consistency evaluation of waste water purification processes at urban water treatment plants. In total, 25 natural surface water samples were collected around St. Petersburg (Russia), analyzed as is, and after ultrasonic treatment. Toxicity of the samples was evaluated using bioassay and MS. Relative errors of toxicity assessment with MS in these samples were below 20%. The system was also applied for fast determination of integral water quality using chemical oxygen demand (COD) values in 20 samples of water from river and ponds in Kolkata (India) and performed with an acceptable precision of 20% to 22% in this task. Furthermore, the MS was applied for fast simultaneous evaluation of COD, biochemical oxygen demand, inorganic phosphorous, ammonia, and nitrate nitrogen at two waste water treatment plants (over 320 samples). Reasonable precision (within 25%) of such analysis is acceptable for rapid water safety evaluation and enables fast control of the purification process. MS proved to be a practicable analytical instrument for various real-world tasks related to water safety monitoring.

A Simple Procedure to Assess Limit of Detection for Multisensor Systems.

Currently, there are no established procedures for limit of detection (LOD) evaluation in multisensor system studies, which complicates their correct comparison with other analytical techniques and hinders further development of the method. In this study we propose a simple and visually comprehensible approach for LOD estimation in multisensor analysis. The suggested approach is based on the assessment of evolution of mean relative error values in calibration series with growing analyte concentration. The LOD value is estimated as the concentration starting from which MRE values become stable from sample to sample. This intuitive procedure was successfully tested with a variety of real data from potentiometric multisensor systems.

In vivo and in vitro application of near-infrared fiber optic probe for Ehrlich carcinoma distinction: Towards the development of real-time tumor margins assessment tool.

This report describes a full-scale experiment on intradermal Ehrlich carcinoma (EC) differentiation in mouse model using NIR spectroscopy in diffuse reflectance mode and chemometric data processing. EC is widely used as an experimental tumor model due to its resemblance with human undifferentiated epithelial tumors and can be applied as a preclinical testing in order to verify the capability of NIR spectroscopy to distinguish cancer from healthy tissues before a clinical research with an aim of creating a new analytical tool for on-line intraoperative tumor margins assessment. The study consists of five steps of NIR spectra measurements: in vivo on the early stage of carcinoma growth; in vivo on the advanced stage of carcinoma growth; in vivo during the surgery; in vitro study of the post-operative materials stored in formalin; in vitro study of the post-operative materials stored in paraffin. It was shown that reliable tumor differentiation with a compact optic fiber probe was possible in all these cases. The classification models were built on two data sets, obtained during in vivo and in vitro measurements; both of them demonstrated 100% specificity and sensitivity.

Critical view on drug dissolution in artificial saliva: A possible use of in-line e-tongue measurements.

Proper monitoring of drug's dissolution is a prerequisite for assessing of taste masking efficacy of pharmaceuticals. Corresponding dissolution procedure is likely to be performed with water. Since the objective of these tests is to examine fate of a pharmaceutical formulation in oral cavity, this choice of solvent seems unsuitable because physical and chemical properties of human saliva are quite far from those of water. Obviously, dissolution profiles registered in water may differ significantly from what really happens with a drug in a mouth cavity. In order to address this discrepancy we examined three different compositions of artificial saliva in dissolution test context in present study. It was found that certain compositions preclude the employment of traditional UV-vis spectroscopy as a detection tool due to strong light scattering in the media caused by viscosity and protein composition modifiers. This issue was circumvented by the use of in-line applied potentiometric multisensor system (e-tongue) and the potential of this new approach for more biorelevant dissolution tests was demonstrated with two model formulations of quinine and ibuprofen.

A sample-effective calibration design for multiple components.

The experimental design of mixtures for multivariate calibration is introduced. The idea of this design is based on uniform distribution of experimental points in a concentration hypercube. Unlike the already reported uniform designs this one is pretty simple and not computationally demanding. The suggested approach does not employ the concept of fixed "levels" and allows for designs with any number of experimental mixtures and any number of components depending on "time and money" considerations for each particular calibration experiment. The performance of the design is assessed with a UV-Vis spectroscopic experiment for simultaneous quantification of four inorganic components in complex mixtures. The performance of the PLS regression models derived from the design is compared with that of cyclic permutation and Kennard-Stone designs. The suggested approach allows for comparable or higher prediction accuracy with the lower number of experimental points.

A combination of dynamic measurement protocol and advanced data treatment to resolve the mixtures of chemically similar analytes with potentiometric multisensor system.

Data processing techniques and measuring protocol are very important parts of the multisensor systems methodology. Complex analytical tasks like resolving the mixtures of two components with very similar chemical properties require special attention. We report on the application of non-linear (artificial neural networks, ANNs) and linear (projections on latent structures, PLS) regression techniques to the data obtained from the flow cell with potentiometric multisensor detection of neighouring lanthanides in the Periodic System of the elements (samarium, europium and gadolinium). Quantification of individual components in mixtures is possible with reasonable precision if dynamic components of the response are incorporated thanks to the use of an automated sequential injection analysis system. The average absolute error in prediction of lanthanides with PLS was around 1 × 10(-4)mol/L, while the use of ANNs allows the lowering of prediction errors down to 2 × 10(-5)mol/L in certain cases. The suggested protocol seems to be useful for other analytical applications where simultaneous determination of chemically similar analytes in mixtures is required.