A new role for PGRP-S (Tag7) in immune defense: lymphocyte migration is induced by a chemoattractant complex of Tag7 with Mts1

PGRP-S (Tag7) is an innate immunity protein involved in the antimicrobial defense systems, both in insects and in mammals. We have previously shown that Tag7 specifically interacts with several proteins, including Hsp70 and the calcium binding protein S100A4 (Mts1), providing a number of novel cellular functions. Here we show that Tag7–Mts1 complex causes chemotactic migration of lymphocytes, with NK cells being a preferred target. Cells of either innate immunity (neutrophils and monocytes) or acquired immunity (CD4⁺ and CD8⁺ lymphocytes) can produce this complex, which confirms the close connection between components of the 2 branches of immune response.     

Investigation of membrane permeability of carp spermatozoa for water molecules

The fundamentals of a photometry method for determination of membrane permeability of some fish spermatozoa for water molecules are presented. Osmotic tolerance of carp spermatozoa membranes was studied using EPR-spectroscopy and photometric analysis methods. It was shown that carp spermatozoa look like the ideal osmometers in their reaction on media of different osmolarity. The value of membrane permeability of carp spermatozoa for water molecules was determined. Data obtained can be used in cryobiology for creating cryoprotective media and regimes of fish sperm cryopreservation.     

Variability of the cytological parameters of Pinus sylvestris L. seeds from the unique Hrenovskoy pine forest

Hrenovskoy pine forest is a unique island stand at the boundary of the species range of Scots pine Pinus sylvestris L. This object is of exceptional economic value, because it serves as a forest-seed base for the Voronezh oblast and some other regions of Russia; therefore, the stand and seed qualities have to be monitored constantly. The results of the first cytogenetic study of the seed progeny of P. sylvestris from the Morozov Grove, a high-quality stand in a reserved site within the Hrenovskoy pine forest, are reported. The studies have been performed in order to obtain a more correct assessment of seed quality based not only on their germination and energy of germination (traditionally used by forest breeders), but also on their genomic stability. The latter may be estimated by the stability of chromosome number in the somatic cells of seedlings and the regularity of mitotic divisions, because they also characterize the state of the generative system of parental forms and may serve as an integrated estimate of the stand development homeostasis. Therefore, the chromosome number, mitotic and nucleolar activities, and the number and spectrum of pathological mitoses (PMs) have been determined. Seedlings have been obtained from 240 seeds (collected from 12 trees) that resulted from free pollination. The cytological analysis of the rootlets of these seedlings has not detected any deviations from the chromosome number typical of the species P. sylvestris L. (2n = 24). However, considerable variation has been found in each family with respect to the mitotic index (MI) (from 4.2 +/- 0.36 to 8.1 +/- 0.39%) and the number of PMs (from 0.5 to 2.1%); micronuclei have also been found in each family (from 0.01 to 0.05%). In general, the phenotypic characteristics and the variation pattern of cytological parameters of the progeny of the trees studied in the Hrenovskoy pine forest, together with the high germination rate of seeds (90-98%), indicate that the current state of pines in the Hrenovskoy pine forest can be considered to be satisfactory. However, the presence of micronuclei in all variants and the decrease in MI in some individual plants (which was, on average, 6.1 +/- 0.03% in the sample studied, i.e., significantly lower than that of P. sylvestris from the Usmanskii pine forest (7.3 +/- 1.2%), where the conditions are optimal for this species) may serve as the bioindicators of the deterioration of both the state of trees themselves and the ecological conditions in the Hrenovskoy pine forest. Therefore, the population structure of this valuable autochtonic stand should be reproduced urgently.     

Identification of signature and primers specific to genus <Emphasis Type="Italic">Pseudomonas</Emphasis> using mismatched patterns of 16S rDNA sequences

Background

Pseudomonas, a soil bacterium, has been observed as a dominant genus that survives in different habitats with wide hostile conditions. We had a basic assumption that the species level variation in 16S rDNA sequences of a bacterial genus is mainly due to substitutions rather than insertion or deletion of bases. Keeping this in view, the aim was to identify a region of 16S rDNA sequence and within that focus on substitution prone stretches indicating species level variation and to derive patterns from these stretches that are specific to the genus.

Results

Repeating elements that are highly conserved across different species of Pseudomonas were considered as guiding markers to locate a region within the 16S gene. Four repeating patterns showing more than 80% consistency across fifty different species of Pseudomonas were identified. The sub-sequences between the repeating patterns yielded a continuous region of 495 bases. The sub-sequences after alignment and using Shanon's entropy measure yielded a consensus pattern. A stretch of 24 base positions in this region, showing maximum variations across the sampled sequences was focused for possible genus specific patterns. Nine patterns in this stretch showed nearly 70% specificity to the target genus. These patterns were further used to obtain a signature that is highly specific to Pseudomonas. The signature region was used to design PCR primers, which yielded a PCR product of 150 bp whose specificity was validated through a sample experiment.

Conclusions

The developed approach was successfully applied to genus Pseudomonas. It could be tried in other bacterial genera to obtain respective signature patterns and thereby PCR primers, for their rapid tracking in the environmental samples.

     

Fermentative hydrolysate of mycelial waste of aminoglycoside antibiotics production as component of culture media used in biosynthesis of tobramycin]

New nutrient media for cultivation of the tobramycin-producing organism were developed. As an additional source of nitrogen the media contain fermentative hydrolysate of the mycelial waste of manufacture of aminoglycoside antibiotics (tobramycin and apramycin). The use of the media provided a 20 to 50% decrease of consumption of soybean meal, an essential food raw material, and design of a low-waste technology for biosynthesis of tobramycin.     

Meiotic analysis of four cross-pollinated generations in a synthetic autotetraploid population of husk tomato (<i>Physalis ixocarpa</i>)

The cultivated husk tomato (Physalis ixocarpa) (2n = 2x = 24) is native from Mexico and Central America and shows a wide genetic variation. Presently, it is the fourth horticultural crop in cultivation surface in Mexico. The working team of this research previously developed an autotetraploid population by using colchicine. The objectives of the present work were to analyze the ploidy level and meiotic behavior of the subsequent generations (C3, C4, C5, C6) from the original (C2) composed only by plants with the duplicated genome from the Rendidora cultivar, and to determine pollen viability. As a diploid control the cultivar Rendidora of P. ixocarpa was used. Ploidy level was determined by flow citometry and meiotic analysis. For the meiotic study, the microsporocytes were prepared by the squash method, stained with carmin and analyzed in diakinesis. Pollen viability was evaluated through 0.01% Buffalo Black staining. The tetraploid condition prevailed through four cross-pollinating generations, maintaining a constant chromosome number 2n = 4x = 48. In diakinesis, the chromosomes of the diploid cultivar were associated into bivalents, whereas in tetraploid plants the chromosomes associated into univalents, bivalents and trivalents. Highly significant differences in bivalent pairing were detected between autotetraploid plants and between generations. Pollen viability did not show significant differences between generations and allowed reproduction. These results indicate that it is possible to develop an autotetraploid cultivar, because the polyploid state is naturally maintained and the plants are fertile. Furthermore, given the differences in bivalent pairing between plants and generations, a response to selection toward meiotic stability is expected.