A Cell Model of Human Small Intestinal Wall Based on Genetically Modified Caco-2 Cells.

We propose a cell model of the human small intestinal wall based on genetically modified Caco-2 cells that allows visualization and quantitative assessment of activation of NF-κB factor and related intracellular pathway by using fluorescence microscopy. A dose-dependent increase in fluorescence intensity of the obtained cells in response to TNFα exposure in concentrations of 1-100 ng/ml was demonstrated. It was found that this parameter correlates with a decrease in the transepithelial resistance of the cell monolayer in response to TNFα and can be used to assess the toxic effects of substances on epithelial cells of the human small intestine.

[Quality control study of engineered skin tissue]. / Metod otsenki kachestva rekonstruirovannogo épidermisa.

OMERO service was used to annotate the cell line HaCaT microscope images by two independent expert groups. The images were obtained in the course of developing tissue-engineered epithelium which consisted of several layers of the keratinocytes. Evaluation of expert opinions was performed by calculation of specificity, sensitivity and accuracy. The best convergence of opinions (91%) was achieved for the confluence of the cell monolayers. Accuracy 70% was observed in determining the extent of cell differentiation after 10 days of incubation. The paper illustrates the usefulness of OMERO service for dynamic cross-validation of quality in the development and standardization of cell preparations.

[Proteome of the human HaCaT keratinocytes: Identification of the oxidative stress proteins after sodium dodecyl sulpfate exposur].

Oxidative stress is a universal response of the skin cell damage of various origins. Sodium dodecyl sulfate (SDS, sodium lauryl sulfate) is an anionic surfactant commonly used as an emulsifying detergent in household cleaners. Sodium dodecyl sulfate is the reference compound for testing toxicity on cellular skin models. The effect of sodium dodecyl sulfate in sub toxic dose 25 µg/mL during 48 h on the protein profile of human keratinocytes HaCaT was studied by tandem mass spectrometry with electrospray ionization. In total, 1064 proteins were found in immortalized human keratinocytes HaCaT, of which about 80% were identified by two or more peptides. The change of the 217 proteins content was revealed, among them 39 according to Gene Ontology are associated with oxidative stress. It has been found that sodium dodecyl sulfate leads to a decrease in the number of proteins/peptides containing carboxymethylated and/or carboxyethylated lysine. We concluded about the promising of the cells redox-balance analysis at testing chemicals in the doses, which do not lead to a decrease in their viability. Possible involvement of sodium dodecyl sulfate in the development of cutaneous neoplasia is discussed.

[Proteomic profiling of HaCaT keratinocytes exposed to skin damaging detergents]. / Proteomnoe profilirovanie keratinotsitov linii HaCaT pri vozdeistvii riada poverkhnostno-aktivnykh veshchestv, vyzyvaiushchikh povrezhdenie kozhi.

The effects of sodium dodecyl sulfate (25 mg/ml) and Triton X-100 (12.5 mg/ml and 25 mg/ml) on the HaCaT immortalized keratinocytes exposed to these surfactants for 48 h were studied. Using shotgun proteomics, a comparative analysis of the proteomic profiles of control and experimental cells after surfactants exposure was carried out. 260 common proteins were identified in control and experimental cells; 33 proteins were found in cells exposed to all three treatments, but not in control cells. These 33 proteins apparently reflect a nonspecific (universal) response of cells to toxic damage by the surfactants. These proteins are associated with activation of cell proliferation, changes in the functional activity of their ER and mitochondria, increased mRNA stability and activation of protein degradation processes in the cells. The possibility of using these proteins as a nonspecific parameter of cell response to cytotoxic damage is discussed. The mass spectrometry proteomics data ("raw", "mgf" and "xml" files) have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifiers PXD007789 and PXD007776.

Correlation between LC50 for Adult Fish and Fish Embryos.

Testing for substance toxicity for living organisms is an important step in the development and adaptation of new drugs for various purposes. Analysis of the dependences between toxicological parameters of chemical substances for various test objects and physicochemical properties of these agents is a promising trend. Partition coefficient logP (logKow) was chosen as the key physicochemical parameter determining the toxicological parameters of the same substance for hydrobionts at different developmental stages. We found a correlation between decimal logarithm of the ratio of LCe50 for fish embryos to LCa50 for adult fish and logP. This dependence was found as a liner combination of equations obtained by drawing a trend line between experimental points and calculation of Pearson's correlation coefficient R.

Changes in the Proteome of HaCaT Keratinocytes Induced by Cytotoxic Substance Triton X-100.

Changes in the proteome of keratinocytes of immortalized HaCaT line exposed to cytotoxic substance Triton X-100 in concentrations of 12.5 and 25 µg/ml were studied by liquid chromatography combined with mass spectrometry. The appearance of proteins involved in the regulation of mitosis, RNA stability, and catabolic processes were detected; the number of apoptosis-associated proteins increased, while the number of proteins involved in differentiation and energy metabolism of keratinocytes decreased.