Nucleolytic activities and appearance of a new DNase in relation to nickel and manganese accumulation inAlyssum murale

Serpentine and non-serpentine plants of Alyssum murale, a nickel (Ni) accumulator plant, from North Greece, were studied in order to examine: (1) The ability of natural plants to accumulate metals; (2) the ability of their seedlings to tolerate increasing concentrations of Ni²⁺ or Mn²⁺ (0, 0.16, 0.32, 0.5 and 1 mmol/L), when grown in nutrient solution; (3) the activities and electrophoretic patterns of root and shoot DNases and RNases under the above conditions. Measurements of metal concentrations in serpentine and non-serpentine natural plants and the respective soils revealed: (1) Very low calcium (Ca)/magnesium (Mg) (0.16) ratio and high concentration of Ni in serpentine soil; (2) very high Ca/Mg (17) ratio and high concentration of manganese (Mn) in non-serpentine soil; (3) the ability of serpentine natural plants to accumulate Ni and the inability of plants of both serpentine and non-serpentine populations to accumulate Mn. A. murale plants grown in nutrient solution with increasing Ni²⁺ or Mn²⁺ concentrations showed a negative correlation between the Ni²⁺ or Mn²⁺ concentrations in the nutrient solution, and the chlorophyll concentration, shoot and especially root length. The accumulation of Ni²⁺ or Mn²⁺ in the plant showed a positive correlation with increasing Ni²⁺ or Mn²⁺ concentrations in the nutrient solution. Application of 0.5 mmol/L Ni²⁺ or Mn²⁺ resulted in the inhibition of DNase activities and the appearance of a new DNase form, in both root and shoot detected by electrophoresis in active ssDNA polyacrylamide gel. The new gel-extracted DNase showed nicking action against plasmid DNA and has been characterised as an endo-DNase. In contrast, electrophoretic patterns and RNase activities were unaffected. According to our studies on growth, both serpentine and non-serpentine plants of A. murale have a constitutive ability to tolerate and accumulate Ni²⁺ or Mn²⁺; they have similar DNase and RNase electrophoretic patterns and show a new DNase form under Ni²⁺ or Mn²⁺ stress. This is the first report on the response of nucleolytic enzymes under metallic elements hyperaccumulation.     

Seasonal and spatial variability of elemental concentrations in boreal forest larch foliage of Central Siberia on continuous permafrost

We measured the seasonal dynamics of major and trace elements concentrations in foliage of larch, main conifer species of Siberia, and we analyzed cryogenic soils collected in typical permafrost-dominated habitats in the Central Siberia. This region offers a unique opportunity to study element fractionation between the soil and the plant because of (i) the homogeneous geological substratum, (ii) the monospecific stands (Larix gmelinii) and (iii) the contrasted habitats (North-facing slope, South-facing slope, and Sphagnum peatbog) in terms of soil temperature, moisture, thickness of the active layer, tree biomass and rooting depth. The variation of these parameters from one habitat to the other allowed us to test the effects of these parameters on the element concentration in larch foliage considered with high seasonal resolution. Statistical treatment of data on larch needles collected 4 times in 3 locations during entire growing season (June–September) demonstrated that : (1) there is a high similarity of foliar chemical composition of larch trees in various habitats suggesting intrinsically similar requirements of larch tree growth for nutrients, (2) the variation of elemental concentrations in larch needles is controlled by the period (within the growing season) and not by the geographical location (South-facing slope, North-facing slope or bog zone) and (3) there are three groups of elements according to their patterns of elements concentration in needles over the growing season from June to September can be identified: (1): nutrient elements [P, Cu, Rb, K, B, Na, Zn, Ni and Cd] showing a decrease of concentration from June to September similar to the behaviour of major nutrients such as N, P and K; (2): accumulating elements [Ca, Mg, Mo, Co, Sr, Mn, Pb and Cr] showing an increase of concentration from June–July to September; (3): indifferent elements [Al, Zr, Fe, Ba, Ti, REEs (Pr, Nd, Ce, La, Gd, Er, Dy, Tb, Lu, Yb, Tm, Sm, Ho, Eu), Y, Th and U] showing a decrease of concentration from June to July and then an increase of concentration to September. A number of micronutrients (e.g., Cu, Zn) demonstrate significant resorption at the end of growing season suggesting possible limitation by these elements. Although the intrinsic requirement seems to be similar among habitats, the total amount of element stored within the different habitats is drastically different due to the differences in standing tree biomass. The partitioning coefficients between soil and larch appear to be among the lowest compared to other environments with variable plants, soils and climates. Applying the “space for time” substitution scenario, it follows that under ongoing climate warming there will be an increase of the element stock following enhanced above-ground biomass accumulation, even considering zero modification of element ratios and their relative mobility. In this sense, the habitats like south-facing slopes can serve as resultant of climate warming effect on element cycling in larch ecosystems for the larger territory of Central Siberia.     

Significance of penetration and metabolism on topical toxicity of diflubenzuron in Spodoptera littoralis and Spodoptera exigua

Toxicity assays indicated that when diflubenzuron (DFB) was topically applied on larvae of the Egyptian cotton leafworm, Spodoptera littoralis (Boisduval), and the beet armyworm, Spodoptera exigua (Hübner), estimated LD₅₀ values were similar, reaching 0.47 and 0.44 ng/larva, respectively. This study evaluated the importance of the rate of uptake and excretion, and of enzymatic metabolism in building up an insecticidal toxicity after topical application on the insect cuticle in both species. In general, penetration of DFB in S. littoralis was about 2-fold higher than in S. exigua, whereas metabolic breakdown was of minor importance in S. littoralis as compared with S. exigua. The current data indicate the importance of cuticle penetration and discuss that extrapolation from oral feeding toxicity results to topical toxicity data should be evaluated with caution, especially for insect growth regulators used in integrated pest management.     

Rational design of genetically stable, live-attenuated poliovirus vaccines of all three serotypes: relevance to poliomyelitis eradication

The global eradication of poliomyelitis caused by wild-type virus is likely to be completed within the next few years, despite immense logistic and political difficulties, and may ultimately be followed by the cessation of vaccination. However, the existing live-attenuated vaccines have the potential to revert to virulence, causing occasional disease, and viruses can be shed by immunocompromised individuals for prolonged periods of time. Moreover, several outbreaks of poliomyelitis have been shown to be caused by viruses derived from the Sabin vaccine strains. The appearance of such strains depends on the prevailing circumstances but poses a severe obstacle to strategies for stopping vaccination. Vaccine strains that are incapable of reversion at a measurable rate would provide a possible solution. Here, we describe the constructions of strains of type 3 poliovirus that are stabilized by the introduction of four mutations in the 5' noncoding region compared to the present vaccine. The strains are genetically and phenotypically stable under conditions where the present vaccine loses the attenuating mutation in the 5' noncoding region completely. Type 1 and type 2 strains in which the entire 5' noncoding regions of Sabin 1 and Sabin 2 were replaced exactly with that of one of the type 3 strains were also constructed. The genetic stability of 5' noncoding regions of these viruses matched that of the type 3 strains, but significant phenotypic reversion occurred, illustrating the potential limitations of a rational approach to the genetic stabilization of live RNA virus vaccines.     

The Interferon-induced Transmembrane Proteins,IFITM1, IFITM2, and IFITM3 Inhibit Hepatitis C Virus Entry

The interferon-induced transmembrane (IFITM) family of proteins have recently been identified as important host effector molecules of the type I interferon response against viruses. IFITM1 has been identified as a potent antiviral effector against hepatitis C virus (HCV), whereas the related family members IFITM2 and IFITM3 have been described to have antiviral effects against a broad range of RNA viruses. Here, we demonstrate that IFITM2 and IFITM3 play an integral role in the interferon response against HCV and act at the level of late entry stages of HCV infection. We have established that in hepatocytes, IFITM2 and IFITM3 localize to the late and early endosomes, respectively, as well as the lysosome. Furthermore, we have demonstrated that S-palmitoylation of all three IFITM proteins is essential for anti-HCV activity, whereas the conserved tyrosine residue in the N-terminal domain of IFITM2 and IFITM3 plays a significant role in protein localization. However, this tyrosine was found to be dispensable for anti-HCV activity, with mutation of the tyrosine resulting in an IFITM1-like phenotype with the retention of anti-HCV activity and co-localization of IFITM2 and IFITM3 with CD81. In conclusion, we propose that the IFITM proteins act in a coordinated manner to restrict HCV infection by targeting the endocytosed HCV virion for lysosomal degradation and demonstrate that the actions of the IFITM proteins are indeed virus and cell-type specific.     

Analysis of host responses of guinea pigs during Helicobacter pylori infection

Host responses of guinea pigs infected with Helicobacter pylori were investigated. Passaged H. pylori colonised the stomach for up to 13 weeks after infection, but after 1 month the number of bacteria fell sharply. Specific antibodies, predominantly of the IgG2 subtype, were present from week 3 onwards. Antibodies to urease A and flagella were abundant. Severe inflammation of the gastric mucosa and damage to the stomach epithelium was seen. Infiltrates of mononuclear cells and eosinophils were found near the parietal glands. As infection progressed, inflammation and tissue damage became more localised and more variable between individual animals. These parameters can be used as markers for colonisation of the stomach by H. pylori.     

Modular neural networks: a survey.

Modular Neural Networks (MNNs) is a rapidly growing field in artificial Neural Networks (NNs) research. This paper surveys the different motivations for creating MNNs: biological, psychological, hardware, and computational. Then, the general stages of MNN design are outlined and surveyed as well, viz., task decomposition techniques, learning schemes and multi-module decision-making strategies. Advantages and disadvantages of the surveyed methods are pointed out, and an assessment with respect to practical potential is provided. Finally, some general recommendations for future designs are presented.     

Radiation-induced changes in liver and kidney alkaline phosphatase and esterase of mice

Whole-body exposure of mice to 2 Gy gamma radiation results in an increase in the specific and total activities of esterase and alkaline phosphatase found in the supernatant of liver and kidney. This increase developed at 1 h, reached a maximum at 24 h, and then declined at 48 h after irradiation. The increase in the activity of these enzymes was not accompanied by an increase in protein content. Electrophoretic analysis on acrylamide gel confirmed the results obtained by the quantitative biochemical analysis. An increase in the level and/or number of bands was observed postirradiation.