[The P-Element Has Not Significant Effect on the Drosophila simulans Viability].

Cases of horizontal transfer of transposable elements (TEs) between species are known for the Drosophilidae family. In the middle of the last century, the case of horizontal transfer of the P-element from the Drosophila willistoni to the D. melanogaster was described. A novel P-element invasion into the D. simulans genome from D. melanogaster occurred approximately 10 years ago. Currently, the P-element has spread across all D. melanogaster population and 30% of D. simulans populations in Europe, Africa and America. In this paper, we investigated the presence of the P-element in D. simulans lines caught in different years in three Asian populations (Tashkent, Nalchik and Sakhalin Island). We also examined the physiological characteristics (cytotype, lifespan, fecundity and locomotor activity) of D. simulans lines with and without the P-element to determine the significance of this new mobile element in the genome. The P-element was found in lines isolated from nature after 2012. The number of P-element copies per genome (two-to-three dozen according to fluorescence in situ hybridization data) was greater than in the American and comparable to the African populations. There were signs of intraspecific hybrid dysgenesis for some pairs of lines. However, in general the presence of the P-element did not adversely affect the physiological characteristics. Either adaptation to the new TE occurs very quickly, or the rate of movement of the P-element is so insignificant that its appearance in the genome remains unnoticed.

[P-M hybrid dysgenesis affects juvenile hormone metabolism in Drosophila melanogaster females].

This paper studies the metabolism of the juvenile hormone, which affects gonads functioning in Drosophila melanogasterfemales under P-M hybrid dysgenesis. It is shown that dysgenic females grown at 29°C have increased levels of the juvenile hormone (its degradation and stress reactivity are reduced), which apparently is a compensatory response to ovarian hypoplasia.

[The rate of transposition and the specificity of transposable element insertions are not sufficient to cause gonadal dysgenesis in Drosophila melanogaster].

Data disputing the unique role of transposable elements (TEs) in the induction of intraspecific gonadal dysgenesis (GD) in Drosophila are discussed. Transposable elements (TEs) occupy the fifth part of the genome of Drosophila melanogaster.

[Patterns of transposable elements distribution on the Drosophila melanogaster polytene chromosomes before and after selection for a quantitative trait].

The effect of selection for radius vein length on the distribution of hybridization sites of the P and hobo transposons and the mdgl and mdg2 retrotransposons on polytene chromosomes of Drosophila melanogaster salivary glands was studied. The patterns of these transposable elements (TEs) distribution were polymorphic in both the parental strain and selected strains. The similarity in mdg1 and mdg2 patterns between strains selected in one direction was closer than between strains selected in opposite directions, but the selected strains were closer to each other than to the parental strain regardless of selection direction. No mdg2 hybridization sites that would be absent in the control were found in the selected strains. There were more mdg2 and hobo hybridization sites in the strains selected in the (+) direction than in the (-) direction. The mobility of hobo copies in the strains studied correlated with the presence of its full-sized copy in the genome. The polymorphism of all TEs studied except for mdgl was greater for strains selected in the (+) direction that in the (-) direction. These facts suggest that some TEs migrate over the genome independently of selection, and others are markers of evolutionary events rather than their causes.

[Change in the distribution of transposable elements in isogenic strain--cause or consequence in Drosophila melanogaster selection for quantitative traits?].

The distribution patterns pf hobo transposon and Dm412 retrotransposon hybridization sites on the salivary gland polytene chromosomes from the larvae of Drosophila melanogaster isogenic strain 51, used for analyzing the effect of transposable element (TE) transposition on the selection for quantitative traits, were studied, It was demonstrated that at least half Dm412 hybridization sites were retained 15 years after isogenization; the frequency of Dm412 transposition varied from 2.0 x 10(-4) to 8.8 x 10(-5) depending on whether the appearance of the same hybridization sites in some individuals were regarded as independent events or as a manifestation of the sample heterogeneity. The distribution patterns of hobo hybridization sites in two isofemale strains derived from isogenic strain 51 differed more noticeably: the number of hobo sites in one of the derivative strains was threefold smaller than in another and only a fraction of the sites was common. Within each derivative strain, the TE distribution was uniform, suggesting that inbreeding had no effect on the Dm412 activity in this strain. The rates of change in the distribution patterns of various TEs in strain 51 corresponded to their spontaneous transposition rates. As isogenic strain accumulates the polymorphism in TE distribution without selection, the TEs are more likely to be the markers of selection events than their inducers. Thus, when studying the effects of various environmental factors on TE transposition even in isogenic strains, it is necessary to perform rounds of close inbreeding to reduce the potential polymorphism.

[Polymorphism of yellow locus in Drosophila melanogaster from natural populations].

We studied molecular characteristics of yellow (y; 1-0.0) locus, which determines the body coloration of phenotypically wild-type and mutant alleles isolated from geographically distant populations of Drosophila melanogaster in different years. According to Southern data, restrictions map of yellow locus of all studied strains differ from each other as well as from that of Oregon stock. FISH analysis shows that in the neighborhood of yellow locus in X chromosome neither P nor hobo elements are found in y1-775 stock, while only hobo is found there in y1-859 and y1-866 stocks, only P element in y+sn849 stock, and both elements in y1-719 stock. Thus, all studied mutant variants of yellow are of independent origin. Yellow locus residing at the very end of X chromosome (region 1A5-8 of cytologic map) carries significantly more transposon than retrotransposon-induced mutations, as compared to white locus (regions 3C2). It is possible that transposons are more active than retrotransposons at the chromosomal ends of D. melanogaster.

[Three-dimensional organization of telomeres in the Drosophila melanogaster salivary glands nuclei].

The 3D-FISH was employed to investigate the telomere topology in polytene nuclei of salivary glands of Drosophila melanogaster. The majorities of telomeres in polytene nuclei of salivary glands in Drosophila strain y(2-717) are localized in the nuclear central area and have no contacts with nuclear membrane. In females of this strain, ectopic contacts between telomeres occur at 25 % higher frequency than in males. HeT-A DNA in y(2-717alk3-2) strain, which is a derivative of y(2-717) carrying an inversion between 1D and 13C bands, is found in region 13 of X chromosome. The frequency of ectopic contacts of telomeres in y(2-717alk3-2) males is 10 % higher than that in y(2-717) strain. The number of ectopic contacts can be significantly different in independent experiments, possibly indicating the role of random factors in the contact formation.

[The effect of genetic environment on locus-specific instability of the yellow gene in Uman' population of Drosophila melanogaster].

The effects of genotype of the laboratory strains, C(1)DX, ywf/Y, 23.5 MRF/CyL4, and C(1)DX,yf; pi2, on locus-specific instability in the yellow gene of the strains y(2-717, y(2-715), and y(2-700 ) from Uman' population of Drosophila melanogaster was studied. Crosses of the males from Uman'-derived lines with the C(1)DX, ywf/Y females yielded a cascade of derivatives, mostly consisting of y+ and y2 alleles, while their crosses with the 23.5 MRF/CyL4 and C(1)DX,yf; pi2 females mostly resulted in the appearance of y+ and y(1) derivatives. The genomes of laboratory strains used in the study contained the full-sized hobo elements, which could differ from one another relative to the structure of variable region and affinity to different DNA sequences.

Fluorescence in situ hybridization analysis of hobo, mdg1 and Dm412 transposable elements reveals genomic instability following the Drosophila melanogaster genome sequencing.

The genome of Drosophila melanogaster strain y cn bw sp has been sequenced and the transposable elements insertion sites have been determined. We hybridized fluorescence-labeled probes directed to the hobo transposon, Dm412 and mdg1 retrotransposons to polytene chromosomes and compared the observed sites to those published in the annotated genome sequence. We observed an almost twofold increase in the number of hobo hybridization sites (46 found as compared to 24 annotated sites). There was no evidence that the hobo transposition rate is slowing over the 10-year period. The patterns of Dm412 and mdg1 sites have changed less dramatically since the time of genome sequencing. Three novel Dm412 hybridization sites were detected while 4 out of 30 annotated sites were missing. Only one additional mdg1 site was found, while 1 out of 29 annotated sites has been lost.

[Persistent locus-specific instability of yellow(27-717) and Noth(Uc-1) in Drosophila melanogaster coincides with hobo multiplication].

According to FISH data the presence of multiple hobo element copies in the unstable yellow and Notch loci in y(2-717) and Uc-1 Drosophila melanogaster stocks, respectively, was found. Locus-specific instability in these strains is caused by hobo multiplication in the respective loci and its subsequent recombination with neighboring hobo copies rather than its insertion-excision.

[Behavior of hobo and P transposons in yellow2-717 unstable line of Drosophila melanogaster and its derivatives after crossing with a laboratory strain].

Using fluorescent in situ hybridization technique (FISH), the frequency of hobo and P mobile elements transpositions on X chromosomes from the y2-717, isolated from the Uman' population of Drosophila melanogaster, as well as from its phenotypically normal and mutant derivatives, obtained as a result of crosses the males examined with the C(I)DX, ywf/Y females, was evaluated. It was demonstrated that the maximum frequency of hobo transpositions on X chromosomes of the males from derivative strains, subjected to repeated hobo-dysgenic crosses reached a value of 1.2 x 10(-2) per site per X chromosome per generation. The number of hobo copies in male X chromosomes from derivative strains was 3 times higher than in the original initial strain. Furthermore, the "old" hobo sites remained unchanged. In derivative strains, the frequency of hobo insertions was higher than that of excisions. One of the derivative strains, y1t-717alk3-2, was characterized by high intra-strain instability of hobo element localization. In the y2-717a1k3 and y1t-717alk3-2 strains a large inversion, In(1)1B; 13CD, was described. At the absence of the full-sized P element in the strains involved in crosses, maximum frequency of P element transpositions in the derivative strains reached a value of 1.2 x 10(-2) per site per X chromosome per generation.

[The effect of gamma-radiation on induction of the hobo element transposition in Drosophila melanogaster].

The transposition frequency of the hobo mobile element in four successive generations of Drosophila melanogaster strain y2-717 after an acute gamma-irradiation with a dose of 30 Gr amounted to 7.5 x 10(-4) per site per genome per generation. Under the same conditions, PCR analysis of the genomic DNA of y2-717 flies detected new variants of defective hobo sequence. No changes in the hobo localization and PCR products compared with the control were detected in the case of single irradiation with doses of 3 and 30 Gr. The localizations of hobo element on polytene chromosomes of y2-717 strain did not change during 11 generations after five exposures of flies to 30 Gr. Irradiation of a highly unstable D. melanogaster strain y+743 did not increase the number of families with mutant progeny, yet increased the total number of mutant descendants almost twofold, from 5 to 9%.

[Transposition of the hobo element in Drosophila melanogaster somatic cells].

Somatic mutation and recombination test on wing cells of Drosophila melanogaster showed that the recombination frequency in the somatic tissues of strains studied correlated with the presence of a full-length copy of the hobo transposable element in the genome. Transposition of hobo in somatic tissue cells at a frequency 3.5 x 10-2 per site per X chromosome was shown by fluorescence in situ hybridization with salivary gland polytene chromosomes of larvae of one of the D. melanogaster strains having a full-length hobo copy.

Babesia DNA detection in canine blood and Dermacentor reticulatus ticks in southwestern Siberia, Russia.

Babesia infection was studied in 21 blood samples of dogs with symptoms of babesiosis and among 72 Dermacentor reticulatus and 70 Ixodes persulcatus ticks from southwestern Siberia, Russia. Babesia DNA was detected by hemi-nested PCR based on the 18S rRNA gene with subsequent direct sequencing. All of the analyzed canine blood samples and three D. reticulatus, but none from I. persulcatus ticks studied were shown to contain Babesia DNA. Nucleotide sequences of the Babesia 18S rRNA gene fragment of 354 bp long for all 24 positive samples appeared to belong to the subspecies Babesia canis canis and differed only at three positions. The Babesia nucleotide sequences from 17 canine blood samples and from one D. reticulatus tick were identical to each other and to previously known B. canis canis from canine blood in Slovenia. Four canine blood samples and the second tick sample contained a mixture of two nucleotide sequences previously found in canine blood. B. canis canis nucleotide sequence from the third tick differed in the unique nucleotide transition and could correspond to a new genetic variant. Thus, the main etiological agent of canine babesiosis in Novosibirsk region is B. canis canis, and D. reticulatus, but not I. persulcatus, ticks could serve as a vector of this infectious agent. To our knowledge, this is the first report of the B. canis canis nucleotide sequences from ticks.

[Cause for maintaining high instability at the yellow gene in Drosophila melanogaster lines, isolated during the "mode for mutation" period in Uman populations]. / Prichina sokhraneniia vysokoi nestabil'nosti po genu yellow v liniiakh Drosophila melanogaster, vydelennykh v period "mody na mutatsiiu" v populiatsii Umani.

Mobile genetic elements are responsible for most spontaneous mutations in Drosophila melenogaster. The discovered in the 1980s phenomenon of frequent change of the wild-type yellow phenotype for a mutant one, and vice-versa, in strains of Drosophila melanogaster isolated from the Uman' natural population can be, according to our data, explained by repeated inversions and reinversions of the gene regulatory region located between the two copies of the hobo transport. However, most molecular genetic events accompanying the process can occur without the phenotype change. After several generations, the strains, remaining phenotypically unchanged, can possess different molecular genetic properties with respect to yellow. Using genetically homogenous or isogenic strains for the genetic analysis or for production of the new plant cultivars or animal breeds, geneticists and breeders often face the problem of stability of the strains. In the present study, the mechanism underlying the generation of instability at the yellow locus of D. melanogaster determined by the hobo-induced genome instability is described.

[Identification of chlamydia and (or) chlamydia-like microorganisms by light microscopy confirmed by electron microscopy and fluorescent in situ hybridization]. / Rasprostranennost' khlamidii i (ili) khlamidiepodobnykh mikroorganizmov u zhenshchin po rezul'tatam svetovoi mikroskopii, podtverzhdennym électronno-mikroskopicheski i s pomoshch'iu FISH-gibridizatsii DNK.

Using light microscopy, we have shown that chlamydia and/or chlamydia-like microorganisms are registered in 20-25% of the healthy part of human population, whereas in patients of the same age with gynecological problems these were found in 40-50%. Commonly, the infection was slightly manifested (less than 5% of cells are infected). These results were confirmed in four months but only in heavily infected patients. The light microscope data are confirmed by observations with electron microscopy, and by FISH hybridization of the total chlamydial DNA on cytological preparations with chlamydial inclusions. In some cases, microcolonies revealed by FISH hybridization occupied the majority of the cytoplasm volume. Occasionally, the DNA material was found on the nuclear surface. It seems likely that in heavily infected cells chlamydia are able to penetrate into the perinucular space.

[Activity of embryonic mink genome during diapause (cytogenetic analysis): nucleolar and extranucleolar rna synthesis]. / Activnost' c'mbrional'nogo genoma norki vo vremia diapauzy (tsitogeneticheslii analiz): iadryshkovoi i vneiadryshkovoi sintez rnk.

The nucleolar and extranucleolar RNA synthesis was studied in the mink blastocysts at different stages of embryonic diapause and during the periimplantation period using cytoradioautography. The data obtained suggest a differential and stage specific activity of the embryonic mRNA and rRNA synthesis during the period of delayed implantation.

[Comparative analysis of methods of diagnosis of chlamydia infection]. / Sravnitel'nyi analiz metodov diagnostiki khlamidioza.

In case of a correct sampling, the diagnostic value of optic and electron microscopy for detecting Chlamydia infection is not inferior to that of direct microimmunofluorescence (DMIF) and higher than that of enzyme immunoassay (EIA). Optic microscopy showed that basal vaginal epithelium and buccal mucosa can be infected with Chlamydia. Provazek bodies were detected in the buccal mucosa of the overwhelming majority of patients with genital chlamydiasis. These results were confirmed by DMIF and EIA. Since none of the diagnostic methods is 100% reliable, we recommend using two methods: inexpensive optic microscopy and polymerase chain reaction or DMIF.

["Mode for mutation" in the natural population of Drosophila melanogaster from Umani is caused by distribution of a hobo-induced inversion in the regulatory region of the yellow gene]. / "Moda na mutatsiiu" v prirodnoi populiatsii Drosophila melanogaster Umani vyzvana rasprostraneniem indutsirovannoi homo-élementom nestabil'noi inversii reguliatornoi chasti gena yellow.

A mutation outburst of the yellow gene occurred in a Drosophila melanogaster population from the town of Uman' from 1982 to 1991 and was associated with the instability of several alleles. Molecular genetic analysis revealed a deletion variant of the hobo transposable element in the same site of the regulatory region of yellow in the mutant alleles and their derivatives. The outburst of the yellow-2 mutations was attributed to the spreading of the X chromosome, which contained an inversion of the yellow regulatory region, through the population. Reinversion resulted in the wild-type phenotype. Crossing lines carrying the inversion with laboratory line C(1)DX, ywf induced instability of the yellow alleles, which was associated with duplication or multiplication of a fragment of the yellow gene. Most derivative lines eventually became stable. The loss of instability was not associated with phenotypic changes; molecular genetic changes included a loss of the duplicated sequences or a deletion of the inverted regulatory region of the yellow gene.

hobo-induced rearrangements are responsible for mutation bursts at the yellow locus in a natural population of Drosophila melanogaster.

In 1981 recurrent local bursts of mutability of the yellow gene were observed in a natural population of Drosophila melanogaster from Uman' (Ukraine). A series of y2-like mutations in the yellow gene were recovered during the period 1982 to 1991. Most of the mutants display the y2-phenotype, i.e. mutant yellow color of wings and body cuticle. Ninety-nine y2 mutants were shown to be generated by an inversion that occurred between two hobo elements, one located 129 bp from the start site of yellow transcription, and the other in the distal telomere region. The y2 phenotype was caused by the separation of the body and wing enhancers from the transcription unit. Many of the y2-like alleles were highly unstable and reverted to y+, which again, gave rise to y2-like mutants. We found that the y2-->y+-->y2 transitions were generated by repeated inversions between the two hobo elements mentioned. The y2 and y+ alleles lost their instability after deletion of the hobo element present at the tip of the X chromosome.