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Correct determination of the instantaneous level and changes of relevant proteins inside individual cells is essential for correct interpretation and understanding of physiological, diagnostic, and therapeutic events. Thus, single-cell analyses are important for quantification of natural cellular heterogeneity, which cannot be evaluated from averaged data of a cell population measurements. Here, we developed an original highly sensitive and selective instrumentation and methodology based on homogeneous single-step bioluminescence assay to quantify caspases and evaluate their heterogeneity in individual cells. Individual suspended cells are selected under microscope and reliably transferred into the 7 µL detection vials by a micromanipulator. The sensitivity of the method is given by implementation of photomultiplying tube with a cooled photocathode working in the photon counting mode. By optimization of our device and methodology, the limits of detection and quantitation were decreased down to 2.1 and 7.0 fg of recombinant caspase-3, respectively. These masses are lower than average amounts of caspase-3/7 in individual apoptotic and even non-apoptotic cells. As a proof of concept, the content of caspase-3/7 in single treated and untreated HeLa cells was determined to be 154 and 25 fg, respectively. Based on these results, we aim to use the technology for investigations of non-apoptotic functions of caspases.

We report the preparation and results of vibrational spectral analysis, which were obtained using both FT-IR and FT-Raman spectroscopy, for three 1-alkyl-1-methylpiperazine-1,4-diium salts (AMPSs), where alkyl is benzyl 4a, n-octadecyl 4b, and methyl 4c, respectively. These were prepared by multistep synthesis from piperazine. The acid base study of AMPSs was performed and corresponding acid base constants were obtained. Single crystals of AMPSs suitable for XRD-analysis were obtained and analyzed. The complete vibrational assignments of wavenumbers were made on the basis of a potential energy distribution. The HOMO and LUMO analysis was used to determine the charge transfer within the molecules. The calculated first hyperpolarizabilities of AMPSs 4a-4c are 48.34, 57.77 and 123.41 times that of urea. As can be seen from the MEP plots, the negative electrostatic potential regions are mainly localized over the chlorine and oxygen atoms for compounds 4a and 4b and chlorine and iodine atoms of compound 4c, and are possible sites for electrophilic attack. (C) 2015 Elsevier B.V. All rights reserved.

We report the vibrational spectral analysis was carried out using FT-IR and FT-Raman spectroscopy for 1-(pyrid-4-yl)piperazine (PyPi). Single crystals of PyPi suitable for X-ray structural analysis were obtained. The acid-base properties are also reported. PyPi supported on a weak acid cation-exchanger in the single protonated form and this system can be used efficiently as the solid supported analogue of 4-N,N-dimethyl-aminopyridine. The complete vibrational assignments of wavenumbers were made on the basis of potential energy distribution. The HOMO and LUMO analysis is used to determine the charge transfer within the molecule and with the molecular electrostatic potential map was applied for the reactivity assessment of PyPi molecule toward proton, electrophiles and nucleopholes as well. The stability of the molecule arising from hyper-conjugative interaction and charge delocalization has been analyzed using NBO analysis. The calculated first hyperpolarizability of PyPi is 17.46 times that of urea. (C) 2013 Elsevier B.V. All rights reserved.

Introduction and aim: In the literature there is evidence that ballet dancers excessively load themedial edge of their foot during the gait in comparison with non-dancers [1]. It can be explained by the dancing positions (e.g. turnout) in which the dancers have a tendency to load the medial edge of foot with greater intensity [2]. The dancers transfer this stereotype into their everyday activities, such as gait. For a successful prevention of possible excessive foot inversion and eversion rehabilitation can be used. The aim of the present study is to determine the influence of rehabilitation intervention on the plantar pressure distribution in professional dancers. Materials andmethods: Seventeen professional ballet dancers (six men, 11 women; mean age: 25.2±5.2 years; mean height: 170.7±9.9 cm; mean weight: 60.0±12.3 kg) participated in the present study. Theirmean dancing experiencewas 16.1±4.8 years. The intensity of dancing training was 9h a day. Foot pressure data was collected using a 2m pressure plate (RSscan International, Olen, Belgium). Eight gait cycles were used for data analysis. Using the Footscan gait software (version 7.97) the foot was divided into ten parts. Dancerswere randomly divided into two groups – experimental (EG) (9) and control group (CG) (8). They underwent two measurements (before and after rehabilitation). The EG has been undergoing six weeks of rehabilitation, the CG has been without intervention. For rehabilitation we were using soft tissues treatment of foot and lower limb and methods based on the scientific principles of developmental kinesiology and the neurophysiological aspects of the maturing locomotor system. Results: The results showed significantly higher values (p