ARC-1, a sequence element complementary to an internal 18S rRNA segment, enhances translation efficiency in plants when present in the leader or intercistronic region of mRNAs

The sequences of different plant viral leaders with known translation enhancer ability show partial complementarity to the central region of 18S rRNA. Such complementarity might serve as a means to attract 40S ribosomal subunits and explain in part the translation-enhancing property of these sequences. To verify this notion, we designed β-glucuronidase (GUS) mRNAs differing only in the nature of 10 nt inserts in the center of their 41 base leaders. These were complementary to consecutive domains of plant 18S rRNA. Sucrose gradient analysis revealed that leaders with inserts complementary to regions 1105–1114 and 1115–1124 (‘ARC-1’) of plant 18S rRNA bound most efficiently to the 40S ribosomal subunit after dissociation from 80S ribosomes under conditions of high ionic strength, a treatment known to remove translation initiation factors. Using wheat germ cell-free extracts, we could demonstrate that mRNAs with these leaders were translated more than three times more efficiently than a control lacking such a complementarity. Three linked copies of the insert enhanced translation of reporter mRNA to levels comparable with those directed by the natural translation enhancing leaders of tobacco mosaic virus and potato virus Y RNAs. Moreover, inserting the same leaders as intercistronic sequences in dicistronic mRNAs substantially increased translation of the second cistron, thereby revealing internal ribosome entry site activity. Thus, for plant systems, the complementary interaction between mRNA leader and the central region of 18S rRNA allows cap-independent binding of mRNA to the 43S pre-initiation complex without assistance of translation initiation factors.     

Biological properties of bacteriophages, active to Yersinia enterocolitica

The biological properties of 16 clones of Y. enterolitica bacteriophages were tested to select the most active for subsequent use. For the first time Y. enterocolitica virulent phages belonging to the family of Podoviridae were described and 7 serological groups of phages with no cross reactions were registered. The technology for the production of new therapeutic and prophylactic Y. enterocolitica polyvalent bacteriophage under laboratory conditions was developed. The effective multiplicity of contamination ensuring the maximum release of phages from bacterial cells, the optimum incubation temperature and the time of exposure were established. The experimental batches of therapeutic and prophylactic Y. enterocolitica polyvalent bacteriophage thus obtained met the requirements for antibacterial preparations.     

Interaction between structure of plant steroids and their effect on phytonematodes

The effects of certain plant steroids of the groups of furostanol glycosides, glycoalkaloids, and alpha-ecdysone on growth and development of phytoparasitic nematodes were studied. It was shown using an experimental system including tomato, Lycopersicon esculentum Mill., and root-knot nematode, Meloidogyne incognita Kofoid et White, that steroid molecule had significant nematicidic activity if it contained a carbohydrate moiety and an additional heterocycle in the steroid core. The maximum nematicidic activity is inherent in glycosides containing chacotriose as the carbohydrate moiety of the molecule. Some compounds tested in this work could be used for protecting plants against phytoparasitic nematodes.     

Fine structure of the female reproductive system in Sobolevicanthus gracilis and Cloacotaenia megalops (Cestoda: Cyclophyllidea)

The fine structure of the female reproductive organs and ducts in two cyclophyllidean cestodes was studied by transmission electron microscopy. All the studied ducts (vitelline, fertilization and vaginal ducts, and oviducts) as well as the uterine and ootype walls have the luminal surface elevated to form apical cytoplasmic lamellae and are surrounded by bands of circular muscules, which are attached to the basement layer. The structure of the studied ducts epithelium corresponds to the pattern described in other ducts of the cestode genital system, namely a nucleate syncytial layer. An exception is the vagina Sobolevicanthus gracilis, in which the surface is lined with not numerous atypical microtriches. The comparison of our results and the data reported for other species is given. It is found out that in different species of cestodes forming oligolecital eggs, there are observed various uterine structures and diverse contacts between the eggs capsules and the uterine epithelium. The formation of special structures and contacts is the evolutionary way from the extrauterine type to more progressive intrauterine type of embryo development in cestodes.     

Psychophysiologic parameters of reproducing emotions in healthy subjects and patients during the first episode of depression

We analyzed the spectral power of EEG rhythms and some psychological features in patients with depression (first episode) and in healthy subjects. In the control group, the distributions of the spectral power of the alpha3 and gamma rhythms was symmetrical. In reproducing joy, the power of the alpha3 rhythm increased in the occipital and that of the gamma rhythm increased in the temporal areas of the left hemisphere. In depressive patients, predominant activation of the left posterior cortical quadrant in the alpha3 band was observed both in the baseline activity and during reproducing emotions of both valences. In these patients, the gamma rhythm predominated in the anterior regions of the right hemisphere during rest. The mental reproduction of joy and grief in depressive patients shifted the maximal gamma rhythm spectral power to the occipital cortical regions. Thus, in depressive patients, any emotional load, regardless of its valence, results in a decrease in the pathologic gamma activity and makes the distribution of cortical electric activity more close to that of healthy subjects.     

Induction of Reactive Oxygen Species and Phytoalexins in Onion (Allium cepa) Cell Culture by Biotic Elicitors Derived from the Fungus Botrytis cinerea

Metabolites of a phytopathogenic fungus Botrytis cinerea Pers. were analyzed for the presence of biotic elicitors. Three groups of elicitors competent in inducing defense responses inAllium cepa cells were identified and partly purified. The recognition of the elicitor signal in onion cells was shown to elevate the concentration of reactive oxygen species (ROS), namely, superoxide anion-radical (O₂^{\overset{-}.}) and hydrogen peroxide (₂₂). The intensity of ROS release depended on chemical identity of elicitor and its concentration. The most active ROS production in onion cells was induced by a protein fraction isolated from the medium for fungus culturing. The carbohydrate elicitors extracted from the fungus cytoplasm and cell walls of mycelia were much less effective. The dynamics of ROS generation comprised two stages. The first stage represented fast and low-amplitude changes that peaked in 15 min after the elicitor treatment. The second stage was more durable and extensive; it occurred in 1.5–6 h after the treatment.     

Distribution of transposons Tn<Emphasis Type="Italic">5044</Emphasis> and Tn<Emphasis Type="Italic">5070</Emphasis> with noncanonical <Emphasis Type="Italic">mer</Emphasis> operons in environmental bacterial populations

The distribution of noncanonical mercury resistance transposons, Tn5044 and Tn5070 , was examined. A characteristic feature of Tn5044 is temperature sensitivity of its mercury operon and the presence in the mer operon of the gene homologous to RNA polymerase subunit. Structural organization of mercury operon Tn5070 , containing minimum gene set (merRTPA), differs from mer operons of both Gram-negative and Gram-positive bacteria. None of more than two thousand environmental bacterial strains displaying mercury resistance and isolated from the samples selected from different geographical regions hybridized to Tn5044- and Tn5070-specific probes. A concept on the existence of cosmopolite, endemic, and rare transposons in environmental bacterial populations was formulated.Translated from Genetika, Vol. 40, No. 12, 2004, pp. 1717–1721.Original Russian Text Copyright © 2004 by Gorlenko, Kalyaeva, Bass, Petrova, Mindlin.     

Heat-induced activation of reducing properties of of sea water

It was shown using the Ellman's reagent that chloride and bicarbonate anions are the heat-induced reducing agents of sea-water, and their combined action is more than additive. Sulfate anions do not exhibit these properties. The influence of sea-water anions on the heat-induced production of hydrogen peroxide was studied by enhanced chemiluminescence in a peroxidase-luminol-p-iodophenol system. In NaCl and NaHCO3 solutions, at concentration and pH values equal to those of sea-water, the production of H2O2 upon heating increased, as compared with water, whereas sulfate anions depressed its formation. By using coumarin-3-carboxylic acid as a fluorescent detector of OH radicals, a substantial increase in the production of radicals in the presence of chloride and bicarbonate anions upon heating was shown. The effect is due to the electron donor properties of these anions, which lead to the decomposition of H2O2 with the formation of OH radicals. The results obtained were considered from the viewpoint of the equivalence of heat and electromagnetic radiation of an absolutely black body. It is supposed that the high-energy quanta of its spectrum lead to the dissociation of anions with the formation of a hydrated electron and radicals. Then a recombination of radicals with the formation of various molecular products takes place.     

Analysis of mutations in the RET proto-oncogene in patients with medullary thyroid tumor

The spectrum of mutations of the RET protooncogene was analyzed in Russian patients with inherited or sporadic medullary thyroid carcinoma (MTC). Four RET exons (11, 13, 15, and 16) were subjected to molecular analysis, and mutations were revealed and identified in 47.4% (9/19) patients with sporadic MTC. In total, six mutations (including three new ones) were observed. The most common mutation affected codon 918 to cause substitution of methionine with threonine and accounted for 31.6% alleles. Analysis of exons 11 and 16 revealed four mutations in patients with inherited multiple endocrine neoplasia type 2 (MEN 2). Mutations were found in each patient. Thyroidectomy was performed in four asymptomatic carriers of RET mutations from three MET 2A families (in two families, affected relatives had bilateral pheochromocytoma). In two patients, analysis of the surgery material revealed MTC microfoci in both lobes of the thyroid gland. The results provide the ground for constructing a bank of genetic information on Russian MTC patients with the clinically verified diagnosis.