Segments of Escherichia coli genome similar to the exons of human prothymosin alpha gene.

Identification of the putative prothymosin alpha homolog in Escherichia coli cells prompted the search for a prothymosin alpha-coding gene in the E. coli genome. A set of interspersed DNA segments was identified, which match various parts of the human prothymosin alpha molecule. Their location in the E. coli genome and high degree of similarity with the appropriate regions of the human prothymosin alpha gene suggest that some kind of trans-splicing should exist in E. coli, which could be responsible for bringing these putative bacterial prothymosin alpha-coding exons together.     

Phytochemical review of the lichen genus <Emphasis Type="Italic">Stereocaulon</Emphasis> (Fam. Stereocaulaceae) and related pharmacological activities highlighted by a focus on nine species

The Stereocaulon genus is one of the fruticose lichen groups distributed worldwide from tropical zones to polar zones. However, the scientific study of this tricky genus is still limited, making it a challenge to study the group further. Detailed morphological studies are essential to discriminate closely shaped species which is illustrated through personal data focused on phyllocladia, apothecia and spores of nine species. Secondary metabolites isolated from Stereocaulon species are mostly some depsides, depsidones, diphenylethers and dibenzofurans which can have a taxonomic value. The use of Stereocaulon lichens as a traditional medicine in several regions of the world and pharmacological studies of extracts and isolated compounds have been compiled. Biological activities as cytotoxic, anti-inflammatory, antibacterial, antifungal or antioxidant are reported.     

Necroptosis is preceded by nuclear translocation of the signaling proteins that induce it

A signaling pathway that induces programmed necrotic cell death (necroptosis) was reported to be activated in cells by several cytokines and various pathogen components. The major proteins participating in that pathway are the protein kinases RIPK1 and RIPK3 and the pseudokinase mixed lineage kinase domain-like protein (MLKL). Recent studies have suggested that MLKL, once activated, mediates necroptosis by binding to cellular membranes, thereby triggering ion fluxes. However, our knowledge of both the sequence of molecular events leading to MLKL activation and the subcellular sites of these events is fragmentary. Here we report that the association of MLKL with the cell membrane in necroptotic death is preceded by the translocation of phosphorylated MLKL, along with RIPK1 and RIPK3, to the nucleus.Apart from the apoptotic cell death pathway that ligands of the tumor necrosis factor (TNF) family can activate, these ligands and various other inducers, including the interferons and various pathogen components, have in recent years been found also to trigger a signaling cascade that induces programmed necrotic death (necroptosis). This cascade encompasses sequential activation of the protein kinases RIPK1 and RIPK3 and the pseudokinase mixed lineage kinase domain-like protein (MLKL).^{1, 2, 3, 4, 5} RIPK3-mediated phosphorylation of MLKL triggers its oligomerization, which is necessary and sufficient for the induction of cell death,^{6, 7, 8} and can also trigger some non-deadly functions.⁹ MLKL was recently suggested to trigger cell death by binding to cellular membranes and initiating ion fluxes through them.^{6, 7, 8, 10} However, its exact molecular target in death induction is contentious.^{6, 8, 10, 11, 12} Current knowledge of the subcellular sites of MLKL action is based mainly on determination of the location of this protein close to the time of cell death. Here we present a detailed assessment of the cellular location of MLKL at different times following its activation. Our findings indicate that before cell death, MLKL translocates to the nucleus along with RIPK1 and RIPK3.     

Interaction of chloramphenicol tripeptide analogs with ribosomes

Chloramphenicol amine peptide derivatives containing tripeptide fragments of regulatory “stop peptides”–MRL, IRA, IWP–were synthesized. The ability of the compounds to form ribosomal complexes was studied by displacement of the fluorescent erythromycin analog from its complex with E. coli ribosomes. It was found that peptide chloramphenicol analogs are able to bind to bacterial ribosomes. The dissociation constants were 4.3-10 μM, which is 100-fold lower than the corresponding values for chloramphenicol amine–ribosome complex. Interaction of the chloramphenicol peptide analogs with ribosomes was simulated by molecular docking, and the most probable contacts of “stop peptide” motifs with the elements of nascent peptide exit tunnel were identified.     

Bimolecular fluorescence complementation based on the red fluorescent protein FusionRed

The aim of this work is to construct split variants of the red fluorescent protein FusionRed, each of which consists of two separate polypeptides, nonfluorescent parts of FusionRed, that can form functional fluorescent proteins upon reassociation. At the first stage, various circularly permuted FusionRed variants have been created (in circular permutants the protein polypeptide chain is divided into two parts, which change places so that the C-terminal part is followed by the N-terminal part). Two variants with the highest rate of chromophore maturation (fluorescence development) have been selected out of 23 tested permutation points. These proteins called cpFR76-73 and cpFR189-188 (the first number indicates the last amino acid residue of the N-terminal part; the second number, the first residue of the C-terminal part) are spectrally similar to parental FusionRed but possess lower fluorescence quantum yields. Split variants corresponding to these two circular permutants have been tested in mammalian cells. For reassembly of the fluorescent protein fragments, heterodimerizing leucine zippers have been used. It has been shown that split variant FR189-188 matures at 37°C and possesses fluorescence brightness similar to that of FusionRed. Consequently, FR189-188 is potentially suitable for a wide range of applications, for example, the study of protein–protein interactions or visualization of cell populations, in which two target gene promoters are simultaneously active.     

Allele polymorphism analysis of hemostasis and folate metabolism genes by real-time microchip PCR

Genetic diagnostics is widely used for detection of risk factors of hereditary thrombophilias caused by molecular defects in the coagulation system. The hereditary thrombophilias are frequently associated with higher incidences of point mutations in hemostasis (F2 20210G>A, F5 1691G>A) and folate metabolism (MTHFR 677C>T, MTHFR 1298A>C) genes. Combinations of gene abnormalities in F2 and/or MTHFR with Leiden mutation (F5 1691G>A) significantly increase risk of thrombosis. Thus, simultaneous analysis of allele polymorphism of these genes is of clinical importance. This study has demonstrated high efficiency of microchip-based multiplex real time PCR for analysis of allele specific polymorphism in hemostasis and folate metabolism genes. Using this test it is possible to analyze polymorphism of the three genes (four point mutations) in a short time; it requires a minimal quantity of DNA template and PCR reagents including DNA polymerase, and thus can be recommended for clinical laboratory diagnostics.     

The phenomenon of the replication of ontogenetic immaturity in adult cats caused by the cerebrospinal fluid of kittens

Studies have been made on the electrical activity of muscles in the hindlimbs in kittens and adult cats before and after injection the liquor from 10-, 21-day and 1-month kittens into the lumbar part of the spinal cord. Analysis was made of the EMG at rest and that of the evoked reactions. It was shown that EMG of adult cats after injection of kitten liquor attained the pattern typical of the donor. The data obtained indicate the presence in kitten liquor of biologically active substances which account for the level of regulatory relationships. Transfer of these substances to adult animals results in the formation of a new level of coordinations of movements which corresponds to immature organism.     

Effect of thyroxine on skeletal muscles in hereditary myodystrophia

The influence of 3-week L-thyroxin administration on contractility and histochemical composition of skeletal muscles was studied in 129Rj mice of normal and dystrophic genotypes. The rise in speed characteristics of isometric contraction and percentage of type II "fast" fibers was established in both types of animals. Contractile properties in dystrophic animals, receiving L-thyroxin, showed a tendency towards normalization and were comparable to those of normal animals.     

Localization of 5' and 3' ends of the ribosome-bound segment of template polynucleotides by immune electron microscopy

Poly(U) with an average chain length of 40-70 nucleotides was modified at the 5'- or 3'-terminal residues with 2,4-dinitrophenyl derivatives. The modified poly(U) was used to form 30S.poly(U) or 70S.poly(U).Phe-tRNA complexes. Localization of the 5' and 3' ends of the template polynucleotide on the 30S subunit and the 70S ribosome was performed by immune electron microscopy using antibodies against dinitrophenyl haptens. The 5' and 3' ends of poly(U) (putative entry and exit sites of the message) were found in the same region both on the 30S subunit and the 70S ribosome. They were located on the dorsal side of the 30S subunit between the head and the body near the groove bordering the side ledge (platform). Comparison of the size of this region with the possible length of the polynucleotide chain covered by the ribosome allowed us to suggest that the message makes a 'U-turn" (or forms a 'loop') as it passes through the ribosome.