The morphology of the apical organ and adjacent epithelium of pilidium prorecurvatum, a pelagic larva of an unknown heteronemertean (Nemertea)

The morphology of pilidia ex gr. recurvatum from Peter the Great Bay (Sea of Japan) was studied by confocal laser scanning and transmission-electron microscopy. The studied pilidium larvae differ from pilidium recurvatum in lacking a posterior ciliary ring and by the presence of a caudal tuft. On this basis, pilidium prorecurvatum is proposed as a new name for the lavae. The apical organ of pilidium prorecurvatum is represented by a thickened epithelium, which consists of uniform columnar monociliary collar cells and is innervated by a pair of serotonergic intraepithelial neurons. The bodies of the serotonergic neurons are located outside of the apical organ, but occasional axons were found at the organ base. The rest of the pilidial epithelium is represented by flattened polygonal multiciliated cells with sparse microvilli; the bodies of two neurons lie in the helmet epithelium immediately adjacent to the apical organ. Morphologically, the apical organ of the pilidium corresponds well to that of other lophotrochozoan larvae, but their homology remains unclear.     

Studies on Disaccharide Nucleoside Synthesis. Mechanism of the Formation of Trisaccharide Purine Nucleosides

Abstract

The unexpected formation of trisaccharide nucleosides during synthesis of purine 5′-O-β-D-ribofuranosylnucleosides in the presence of Lewis acids was observed.     

Synthesis of Dioxolane Analogues of Dideoxynucleotides and Their Substrate Properties in DNA Synthesis Reactions

Abstract

Dioxolane derivatives of dNTP were prepared and their substrate properties were investigated in DNA synthesis reactions.     

Isolation and Purification of Enzymes from Ligninolytic Complex of the Basidial Fungus <Emphasis Type="Italic">Trametes pubescens</Emphasis> (Schumach.) Pilat and Study of Their Properties

A method for purification of enzymes from the ligninolityc complex of the basidiomycete Trametes pubescens (Schumach.) Pilat has been elaborated. Two homogeneous isoforms of laccases (laccase 1 and laccase 2) as well as a homogeneous preparation of lignin peroxidase were isolated. Basic biochemical parameters of the enzymes were determined, such as the molecular weights (67, 67, and 45 kD, respectively), isoelectric points (5.3, 5.1, and 4.2, respectively), as well as content and composition of the carbohydrate moiety of the laccases (N-acetylglucosamine, mannose, and xylose). The pH dependences and thermal stabilities of the laccases were investigated. The kinetic parameters of the enzymatic reactions catalyzed by the laccases were determined using different substrates, such as catechol, hydroquinone, 2,2 -azinobis-(3-ethylbenzthiazoline-6-sulfonate), and K4Fe(CN)6. The structure of the active sites of both laccases and the lignin peroxidase were studied by EPR, CD, and UV-VIS spectroscopy, as well as using fluorescence analysis. Our studies showed similarity of the spectral characteristics of the two laccases, whereas their kinetic properties were found to be different.     

Fusion with the cold-active esterase facilitates autotransporter-based surface display of the 10th human fibronectin domain in Escherichia coli

Cell surface display is a popular approach for the construction of whole-cell biocatalysts, live vaccines, and screening of combinatorial libraries. To develop a novel surface display system for the popular scaffold protein 10th human fibronectin type III domain (¹⁰Fn3) in Escherichia coli cells, we have used an α-helical linker and a C-terminal translocator domain from previously characterized autotransporter from Psychrobacter cryohalolentis K5^T. The level of ¹⁰Fn3 passenger exposure at the cell surface provided by the hybrid autotransporter Fn877 and its C-terminal variants was low. To improve it, the fusion proteins containing ¹⁰Fn3 and the native autotransporter passenger Est877 or the cold-active esterase EstPc in different orientations were constructed and expressed as passenger domains. Using the whole-cell ELISA and activity assays, we have demonstrated that N-terminal position of EstPc in the passenger significantly improves the efficiency of the surface display of ¹⁰Fn3 in E. coli cells.

     

Inactivation of Receptor Tyrosine Kinases Overcomes Resistance to Targeted <Emphasis Type="Italic">B-RAF</Emphasis> Inhibitors in Melanoma Cell Lines

The discovery of B-RAF activating mutations in malignant melanoma cells has led to the development of a number of targeted drugs, which block exclusively the mutant B-RAF protein. Tumor cells often acquire resistance to B-RAF inhibitors via activation of alternative signaling pathways. One of the resistance mechanisms is activation of PDGF, VEGF, c-KIT, and certain other tyrosine kinases. The possibility of overcoming the resistance to the B-RAF inhibitor Vemurafenib by inactivating receptor tyrosine kinases (RTKs) was studied in metastatic melanoma cell lines differing in B-RAF mutations and RTK activity. It was found that RTK inactivation may help to overcome resistance to B-RAF inhibitors via inhibition of tyrosine kinase phosphorylation and a subsequent blocking of the PI3K-AKT-mTOR and MEK-ERK1/2 downstream signaling pathways. The changes eventually mitigated the cell growth and enhanced the Vemurafenibdependent cell cycle arrest.     

Influence of the activation of the immune system cells on the parameters of lipid metabolism in macrophages

The influence of tumor necrosis factor a (TNF-alpha) and media, conditioned by activated macrophages and lymphocytes and containing a complex of biologically active compounds (including cytokines), on the parameters of lipid metabolism in macrophages was studied. The addition of recombinant TNF-alpha and immunocompetent cell-conditioned media to mouse peritoneal macrophages culture stimulated labelled oleate incorporation into cholesterol esters and triglycerides, as well as labelled glycerine incorporation into cholesterol esters, but inhibited labelled cholesterol incorporation into cholesterol esters. One of the mechanisms of the influence of activated immunocompetent cells on cholesterol metabolism in macrophages was, supposedly, the stimulation of sphigmomyelinase activity by a complex of anti-inflammatory cytokines produced by these cells on their activation.