Intracellular Localization and Gene Expression Analysis Provides New Insights on LEA Proteins' Diversity in Anhydrobiotic Cell Line.

Anhydrobiosis, an adaptive ability to withstand complete desiccation, in the nonbiting midge Polypedilum vanderplanki, is associated with the emergence of new multimember gene families, including a group of 27 genes of late embryogenesis abundant (LEA) proteins (PvLea). To obtain new insights into the possible functional specialization of these genes, we investigated the expression and localization of PvLea genes in a P. vanderplanki-derived cell line (Pv11), capable of anhydrobiosis. We confirmed that all but two PvLea genes identified in the genome of P. vanderplanki are expressed in Pv11 cells. Moreover, PvLea genes are induced in Pv11 cells in response to anhydrobiosis-inducing trehalose treatment in a manner highly similar to the larvae of P. vanderplanki during the real induction of anhydrobiosis. Then, we expanded our previous data on PvLEA proteins localization in mammalian cells that were obtained using C-terminal fusions of PvLEA proteins and green fluorescent protein (GFP). We investigated PvLEA localization using N- and C-terminal fusions with GFP in Pv11 cells and the Sf9 insect cell line. We observed an inconsistency of PvLEA localization between different fusion types and different cell cultures, that needs to be taken into account when using PvLEA in the engineering of anhydrobiotic cell lines.

New group of transmembrane proteins associated with desiccation tolerance in the anhydrobiotic midge Polypedilum vanderplanki.

Larvae of the sleeping chironomid Polypedilum vanderplanki are known for their extraordinary ability to survive complete desiccation in an ametabolic state called "anhydrobiosis". The unique feature of P. vanderplanki genome is the presence of expanded gene clusters associated with anhydrobiosis. While several such clusters represent orthologues of known genes, there is a distinct set of genes unique for P. vanderplanki. These include Lea-Island-Located (LIL) genes with no known orthologues except two of LEA genes of P. vanderplanki, PvLea1 and PvLea3. However, PvLIL proteins lack typical features of LEA such as the state of intrinsic disorder, hydrophilicity and characteristic LEA_4 motif. They possess four to five transmembrane domains each and we confirmed membrane targeting for three PvLILs. Conserved amino acids in PvLIL are located in transmembrane domains or nearby. PvLEA1 and PvLEA3 proteins are chimeras combining LEA-like parts and transmembrane domains, shared with PvLIL proteins. We have found that PvLil genes are highly upregulated during anhydrobiosis induction both in larvae of P. vanderplanki and P. vanderplanki-derived cultured cell line, Pv11. Thus, PvLil are a new intriguing group of genes that are likely to be associated with anhydrobiosis due to their common origin with some LEA genes and their induction during anhydrobiosis.

Parylene Based Memristive Devices with Multilevel Resistive Switching for Neuromorphic Applications.

In this paper, the resistive switching and neuromorphic behaviour of memristive devices based on parylene, a polymer both low-cost and safe for the human body, is comprehensively studied. The Metal/Parylene/ITO sandwich structures were prepared by means of the standard gas phase surface polymerization method with different top active metal electrodes (Ag, Al, Cu or Ti of ~500 nm thickness). These organic memristive devices exhibit excellent performance: low switching voltage (down to 1 V), large OFF/ON resistance ratio (up to 104), retention (≥104 s) and high multilevel resistance switching (at least 16 stable resistive states in the case of Cu electrodes). We have experimentally shown that parylene-based memristive elements can be trained by a biologically inspired spike-timing-dependent plasticity (STDP) mechanism. The obtained results have been used to implement a simple neuromorphic network model of classical conditioning. The described advantages allow considering parylene-based organic memristors as prospective devices for hardware realization of spiking artificial neuron networks capable of supervised and unsupervised learning and suitable for biomedical applications.

Bacillus intermedius ribonuclease (BINASE) induces apoptosis in human ovarian cancer cells.

The cytotoxic effects of Bacillus intermedius RNase (binase) towards ovarian cancer cells (SKOV3 and OVCAR5) were studied in comparison to normal ovarian epithelial cells (HOSE1 and HOSE2). Binase decreased viability and induced the selective apoptosis of ovarian cancer cells. The apoptosis rate was 50% in SKOV3 and 48% in OVCAR5 cells after 24 h of binase treatment (50 µg/ml). Binase-induced apoptosis in these cell lines was accompanied by caspase-3 activation and poly(ADP-ribose) polymerase fragmentation. Normal ovarian epithelial cells were not affected by binase, except for a slight decrease of HOSE2 cell viability and the appearance of traces of activated caspase-3, but not the poly(ADP-ribose) polymerase 85-kDA fragment. Binase did not induce alteration of EZH2 (enhancer of zeste-homolog-2) protein expression neither, in tumor nor in normal cells. In conclusion, selective binase-induced cell death and apoptosis via poly(ADP-ribose) polymerase fragmentation may serve as a new treatment option against ovarian cancer progression.