Changes in the Activity and Pattern of Hemolymph Esterases in the Larvae of Greater Wax Moth Galleria mellonellaL. (Lepidoptera, Pyralidae) during Mycosis

The extended pattern of multiple esterase forms has been revealed in the hemolymph of wax moth Galleria mellonellalarvae infected by the fungi Metarhizium anisopliae, Beauveria bassianaor Paecilomyces fumoso-roseus. The total esterase activity of the hemolymph also increases during mycosis. Mechanical damage of the cuticle, treatment with deltamethrin, and chilling of the caterpillars induced similar changes in the hemolymph pattern of esterase activity. Presumably, the changed spectrum and activity of the hemolymph esterases during mycosis is due to the damaged cuticle and epidermis cells.     

A unique,thermostable dimer linkage structure of RD114 retroviral RNA

Retroviruses package their genome as RNA dimers linked together primarily by base-pairing between palindromic stem–loop (psl) sequences at the 5′ end of genomic RNA. Retroviral RNA dimers usually melt in the range of 55°C–70°C. However, RNA dimers from virions of the feline endogenous gammaretrovirus RD114 were reported to melt only at 87°C. We here report that the high thermal stability of RD114 RNA dimers generated from in vitro synthesized RNA is an effect of multiple dimerization sites located in the 5′ region from the R region to sequences downstream from the splice donor (SD) site. By antisense oligonucleotide probing we were able to map at least five dimerization sites. Computational prediction revealed a possibility to form stems with autocomplementary loops for all of the mapped dimerization sites. Three of them were located upstream of the SD site. Mutant analysis supported a role of all five loop sequences in the formation and thermal stability of RNA dimers. Four of the five psls were also predicted in the RNA of two baboon endogenous retroviruses proposed to be ancestors of RD114. RNA fragments of the 5′ R region or prolonged further downstream could be efficiently dimerized in vitro. However, this was not the case for the 3′ R region linked to upstream U3 sequences, suggesting a specific mechanism of negative regulation of dimerization at the 3′ end of the genome, possibly explained by a long double-stranded RNA region at the U3-R border. Altogether, these data point to determinants of the high thermostability of the dimer linkage structure of the RD114 genome and reveal differences from other retroviruses.     

The Relationship between the Structure of the Tick-Borne Encephalitis Virus Strains and Their Pathogenic Properties

Tick-borne encephalitis virus (TBEV) is transmitted to vertebrates by taiga or forest ticks through bites, inducing disease of variable severity. The reasons underlying these differences in the severity of the disease are unknown. In order to identify genetic factors affecting the pathogenicity of virus strains, we have sequenced and compared the complete genomes of 34 Far-Eastern subtype (FE) TBEV strains isolated from patients with different disease severity (Primorye, the Russian Far East). We analyzed the complete genomes of 11 human pathogenic strains isolated from the brains of dead patients with the encephalitic form of the disease (Efd), 4 strains from the blood of patients with the febrile form of TBE (Ffd), and 19 strains from patients with the subclinical form of TBE (Sfd). On the phylogenetic tree, pathogenic Efd strains formed two clusters containing the prototype strains, Senzhang and Sofjin, respectively. Sfd strains formed a third separate cluster, including the Oshima strain. The strains that caused the febrile form of the disease did not form a separate cluster. In the viral proteins, we found 198 positions with at least one amino acid residue substitution, of which only 17 amino acid residue substitutions were correlated with the variable pathogenicity of these strains in humans and they authentically differed between the groups. We considered the role of each amino acid substitution and assumed that the deletion of 111 amino acids in the capsid protein in combination with the amino acid substitutions R16K and S45F in the NS3 protease may affect the budding process of viral particles. These changes may be the major reason for the diminished pathogenicity of TBEV strains. We recommend Sfd strains for testing as attenuation vaccine candidates.     

Development of the microcirculatory bed of the human tongue in the prenatal period

Morphofunctional regularities of formation and development of the blood microcirculatory bed in the human tongue have been studied in the prenatal period of morphogenesis. 119 human embryos and fetuses at the age of 5 weeks--9 months have been investigated. A complex of methods have been used: common histological (hematoxylin--eosin, after van Gieson and Mallory), injection of the lingual vessels with 20% suspension of Indian ink--gelatin, transmissive electron microscopy. General regularities of organogenesis, stages of the blood microcirculatory bed development and peculiarities of the process on formation of the primary protocapillary lingual blood bed are revealed. Regularities in structure of the terminal vascular constructions are studied for each structural element of the organ--mucosal membrane, muscles, glands, lingual tonsil. For these elements at the ultrastructural level certain features of the organic specificity in the structure of the blood microcirculatory bed links are determined.     

Effect of electromagnetic radiation in a decimeter wave-length range on the calcium current of molluscan neurons

Microwave effect on calcium current of the dialysed snail neurons was investigated. Isolated neurons were irradiated by unmodulated and modulated microwaves (900 MHz) with modulation frequency 0,5 divided by 1000 Hz and SAR = 0,1 divided by 20 W/kg. Calcium current increase was shown to be induced by microwave heating. The current increments were proportional to SAR. Microwave effects on the charge distribution of the cell membrane surface and other non-thermal special microwave effects were not detected.