The construction and use of inosine-containing primers for searching for and identifying the genes of insecticidal crystal proteins in Bacillus thuringiensis]

The specific to 3 types of Cry genes primers containing inosine were constructed to find crystal insecticidal protein Bacillus thuringiensis genes using PCR. A number of new B. thuringiensis strains isolated in Ukraine were investigated using these PCR primers. As a results, Cry genes were found, some of them were identified and demonstrated high homology to Cry1Ba2 and Cry1Bc genes.     

Occurrence of amitotic division of trophoblast cell nuclei in blastocysts of the western spotted skunk (Spilogale putorius latifrons)

A cytogenetic examination of spreaded cells of diapausing and early activated blastocysts obtained from 7 female western spotted skunks was performed. Mitosis was not observed in 1626 cells obtained from 9 diapausing blastocysts; however, 12 (1.5%) figures of diploid mitosis were seen in 851 cells from 5 early activated embryos. Diameter of the cell nuclei varied from 4 to 29 microm during diapause, and from 5 to 40 microm in activated blastocyst, and the heterogeneity in nuclear size was significantly different between diapausing and activated embryos (P<0.01). About 80% of nuclei from diapausing blastocysts measured 9 to 16 microm, whereas a similar percentage of nuclei from activated blastocysts ranged from 15 to 27 microm. Many enlarged nuclei exhibited morphological features characteristic of mammalian polytene (i.e. endopolyploid with polytenic organization of chromosomes) trophoblast cells. The number of silver stained nucleoli in all the nuclei did not exceed 2, which corresponds to the number of nucleolus organizers in the diploid karyotype in this species of skunk and suggests the polytene organization of chromosomes in enlarged nuclei. About 10% of large interphase nuclei were observed to undergo amitosis, i.e. direct division by constriction. The resulting nuclear fragments in diapausing blastocysts usually had normal morphology and active nucleoli. In activated embryos, nearly 15% of amitotically divided nuclei appeared to be dividing into fragments of unequal size, one of which had normal cell nuclear morphology and extremely large silver positive nucleoli, and the other fragment exhibited signs of cell death. We interpret these data as indicating that 1) amitotic division of trophoblast endopolyploid cell nuclei in the skunk blastocysts may generate new trophoblast cells which contribute to increased cell number during both diapause and activation stages, and 2) activation of blastocysts after diapause is related to the production of trophoblast cells with enhanced synthetic capabilities.     

Novel efficient promoter of the mitochondrial porin,voltage-dependent anion channel (VDAC), in the genome of the <Emphasis Type=Italic>Yarrowia lipolytica</Emphasis> yeast

This article presents the characteristics of the highly inducible promoter of the gene encoding the mitochondrial porin, the voltage-dependent anion channel (VDAC). This promoter is recommended for use in new genetic constructs both in basic research for assessing the adaptive strategy of lower eukaryotes under adverse conditions and in designing new highly competitive transformants producing economically important compounds (proteins, lipids, and organic acids) on its basis.     

The Ratio between the Frequencies of Two Forms of Amitotic Division of Trophoblast Cell Nuclei in the Mink Blastocysts during the Delayed Implantation Period

A comparative study of amitotic division activity of trophoblast cells by constriction or extrusion in the blastocysts of American mink during the period of obligatory implantation delay was performed. The frequency of occurrence of amitotic figures was found to be approximately 10% upon the resumption of blastocyst growth (the blastocyst size was 0.4 mm in diameter) and nearly 20% at the stage of active growth (0.9 mm) and at the stage of expansion before blastocyst attachment to the uterine wall (1.7 mm). The ratios between the frequencies of division by extrusion and constriction at these three stages were 2 : 1, 5 : 1, and 4 : 1, respectively. We suggest that the cells produced via different forms of amitosis ways play different roles in trophoblast differentiation.     

Germinal mass of the rediae of Trematoda

Dynamic of the reproduction of the trematodes Notocotylus imbricatus (Notocotylidae), Echinostoma caproni (Echinostomatidae), Sphaeridiotrema globulus and Psilotrema tuberculata (Psilostomatidae) parthenites was observed. The formation of generative cells takes place only as a result of undifferentiated cells proliferation and following differentiation. These processes and early stages of embryogenesis are taking place only in the special reproductive organ--germinal mass. The germinal mass is always formed at the posterior part of the body. The process of reproduction had been finishing to the beginning of the generating of cercariaea of new age by parthenites. Rediae of different generations in fact stop producing new generative cells with the beginning of the appearance of a new generation.     

Allo-Reactivity of Mesenchymal Stem Cells in Rhesus Macaques Is Dose and Haplotype Dependent and Limits Durable Cell Engraftment In Vivo

The emerging paradigm that MSCs are immune privileged has fostered the use of “off-the-shelf” allogeneic MSC-based therapies in human clinical trials. However, this approach ignores studies in experimental animals wherein transplantation of MSCs across MHC boundaries elicits measurable allo-immune responses. To determine if MSCs are hypo-immunogeneic, we characterized the immune response in rhesus macaques following intracranial administration of allogeneic vs. autologous MSCs. This analysis revealed unambiguous evidence of productive allo-recognition based on expansion of NK, B and T cell subsets in peripheral blood and detection of allo-specific antibodies in animals administered allogeneic but not autologous MSCs. Moreover, the degree of MHC class I and II mismatch between the MSC donor and recipient significantly influenced the magnitude and nature of the allo-immune response. Consistent with these findings, real-time PCR analysis of brain tissue from female recipients administered varying doses of male, allogeneic MSCs revealed a significant inverse correlation between MSC engraftment levels and cell dose. Changes in post-transplant neutrophil and lymphocyte counts also correlated with dose and were predictive of overall MSC engraftment levels. However, secondary antigen challenge failed to elicit a measurable immune response in allogeneic recipients. Finally, extensive behavior testing of animals revealed no main effect of cell dose on motor skills, social development, or temperament. Collectively, these data indicate that allogeneic MSCs are weakly immunogenic when transplanted across MHC boundaries in rhesus macaques and this negatively impacts durable engraftment levels. Therefore the use of unrelated donor MSCs should be carefully evaluated in human patients.     

Comparative analysis of respiratory activity in the wild type strain of <Emphasis Type=Italic>Neurospora crassa</Emphasis> and its photoreceptor complex mutants

Cell respiratory activity of protoplasts obtained from the wild type of Neurospora crassa and photoreceptor complex WCC—white collar 1 (wc-1) and white collar 2 (wc-2)—mutants of Neurospora crassa strains was investigated. Respiration inhibition by KCN in the presence of 25 mM succinate was similar in all strains and did not exceed 83–85% against control. The significant induction of KCN-resistant respiratory pathway occurred under 1% glucose oxidation in wc-1 and wc-2 mutants if compared with the wild type strains. The inhibitors of the main (cytochrome) pathway of electron transfer in mitochondria—1 mM KCN and antimycin A (4 μg/ml)—blocked the respiration rate of the protoplasts from N. crassa wild type by 75%, while the cell respiration of wc-1 and wc-2 strains was suppressed by approximately 50%. The specific inhibitor of alternative oxidase—10 mM salicylhydroxamic acid (SHAM)—in combination with the blockers of mitochondrial electron transfer chain caused the total suppression of respiratory activity of protoplasts in all studied strains. It is supposed that an increase of KCN-resistance in WCC mutants under glucose oxidation is connected with alternative oxidase activation as the result of failure in reception and signal transduction of active oxygen species.     

Phylogenetic Analysis of Kyrgyz Horse Using 17 Microsatellite Markers

Russian Journal of Genetics - Abstract—The results of this study are the first in assessing the subpopulation subdivision of the Kyrgyz horse breed. Horse genotyping was performed using 17...