[The haplomatch program for comparing Y-chromosome STR-haplotypes and its application to the analysis of the origin of Don Cossacks].

STR haplotypes of the Y chromosome are widely used as effective genetic markers in studies of human populations and in forensic DNA analysis. The task often arises to compare the spectrum of haplotypes in individuals or entire populations. Performing this task manually is too laborious and thus unrealistic. We propose an algorithm for counting similarity between STR haplotypes. This algorithm is suitable for massive analyses of samples. It is implemented in the computer program Haplomatch, which makes it possible to find haplotypes that differ from the target haplotype by 0, 1, 2, 3, or more mutational steps. The program may operate in two modes: comparison of individuals and comparison of populations. Flexibility of the program (the possibility of using any external database), its usability (MS Excel spreadsheets are used), and the capability of being applied to other chromosomes and other species could make this software a new useful tool in population genetics and forensic and genealogical studies. The Haplomatch software is freely available on our website www.genofond.ru. The program is applied to studying the gene pool of Cossacks. Experimental analysis of Y-chromosomal diversity in a representative set (N = 131) of Upper Don Cossacks is performed. Analysis of the STR haplotypes detects genetic proximity of Cossacks to East Slavic populations (in particular, to Southern and Central Russians, as well as to Ukrainians), which confirms the hypothesis of the origin of the Cossacks mainly due to immigration from Russia and Ukraine. Also, a small genetic influence of Turkicspeaking Nogais is found, probably caused by their occurrence in the Don Voisko as part of the Tatar layer. No similarities between haplotype spectra of Cossacks and Caucasus populations are found. This case study demonstrates the effectiveness of the Haplomatch software in analyzing large sets of STR haplotypes.

[Effect of low temperature on the resistance and functional activity of the photosynthetic apparatus of wheat plants].

The dynamics of cold resistance and the activity of the photosynthetic apparatus (PSA) of wheat germs at 4 degrees C were studied. It was shown that in the first hours of cold, a certain functional readjustment to the changed conditions takes place in the plant organism. A decrease in the activity of the PSA and cessation of the linear growth of the leaf are registered at this stage along with an increase in resistance, as well as an increase in the coefficient of nonphotochemical extinguishing of the fluorescence of chlorophyll. In one to four days, when resistance reaches its maximum, photosynthesis and the rate of electron transport are stabilized, the chlorophyll content in the lightcollecting complex increases, and the growth recommences. The final stage of adaptation (days 4-7) is characterized not only by the steady level of resistance but also by new functional organization of the PSA, which allows the plants to endure the lowered temperature successfully.

[Genetic ecological monitoring in human populations: heterozygosity, mtDNA haplotype variation, and genetic load].

Yu. P. Altukhov suggested that heterozygosity is an indicator of the state of the gene pool. The idea and a linked concept of genetic ecological monitoring were applied to a new dataset on mtDNA variation in East European ethnic groups. Haplotype diversity (an analog of the average heterozygosity) was shown to gradually decrease northwards. Since a similar trend is known for population density, interlinked changes were assumed for a set of parameters, which were ordered to form a causative chain: latitude increases, land productivity decreases, population density decreases, effective population size decreases, isolation of subpopulations increases, genetic drift increases, and mtDNA haplotype diversity decreases. An increase in genetic drift increases the random inbreeding rate and, consequently, the genetic load. This was confirmed by a significant correlation observed between the incidence of autosomal recessive hereditary diseases and mtDNA haplotype diversity. Based on the findings, mtDNA was assumed to provide an informative genetic system for genetic ecological monitoring; e.g., analyzing the ecology-driven changes in the gene pool.

[The Russian gene pool: gene geography of Alu-insertions (ACE, APOA1, B65, PV92 TPA25)].

The analysis of five Alu insertion loci (ACE, AP4OA1, B65, PV92, TPA25) has been carried out for the first time in 10 Russian populations (1088 individuals), covered all parts of historical area of the Russian ethnos. Depending on locus, Russian populations exhibit similarity with their western (European populations) or with the eastern (populations of the Ural region) neighbors. Considering frequencies of the studied Alu-insertions, Russian gene pool exhibits low variation: average difference between populations is d = 0.007, whereas on classical markers, mtDNA and Y chromosome heterogeneity of Russian gene pool is essentially higher (0.013, 0.033 and 0.142 respectively). Therefore, this set of five Alu insertions has lower variability on the intra-ethnic level. However in inter-ethnic comparisons the clear pattern was obtained: 13 Eastern European ethnic groups formed three clusters, according with their historical and geographical position--East Slavic, Caucasian and South Ural clusters. The obtained data confirms efficiency of using Alu insertions for studying genetic differentiation and history of a gene pool of the Eastern European populations.

[Changes in the activity of proteases and trypsin inhibitors in wheat leaves in the initial period of cold hardening and subsequent dehardening].

The dynamics of amidase, cysteine protease, and trypsin inhibitor activities were studied in the leaves of wheat (Triticum aestivum L.) seedlings grown under controlled conditions (25 degrees C, illuminance 10 kLx, 14-h photoperiod) and subjected to cold hardening (5 degrees C, 10 kLx, 14-h photoperiod). Changes in the activity of amidases and cysteine proteases proved to precede an increase in cold resistance during cold hardening and a decrease in cold resistance after the end of cold hardening. The activity of trypsin inhibitors changed only during cold hardening. It is suggested that amidases, cysteine proteases, and trypsin inhibitors are involved in the cold adaptation of plants.

[Periapical tissues microvessels reactivity study after pulp extirpation and on different stages of endodontic treatment].

Reaction of the microvessels in the gingival periapical tissues on pulp extirpation was followed by hyperemia development in microcirculatory bed, its expression increased in proportion as parodontal inflammation intensified and further strengthened after root canal filling. Microcirculation normalization in the gingival periapical tissues after endodontic treatment of chronic pulpitis and pulp extirpation as part of endodontic treatment was evident in 1 month in intact parodontium and in 6 months in case of its inflammation.

[Effect of prednisolon on trachea smooth muscle of normal rats and rats with fibrosing alveolitis].

Effect of prednisolone on isolated preparations of trachea of normal rats and rats with fibrosing alveolitis was studied. Prednisolone at a concentration of 0.4 microg/l decreased responses of smooth muscle on stimulation of preganglionar nerve fibers at trachea areas with intramural ganglia in rats with acute alveolitis by 48%, while in normal rats--by 19% of control. In trachea preparations without ganglia, prednisolone at a dose of 10 microg/l decreased responses of muscle to the nerve fiber stimulation by 21.3%. The higher prednisolone doses were less efficient: 0.1-10 microg/l glucocorticoid practically did not affect the smooth muscle responses produced by stimulation of muscle cells. In rats with fibrosing alveolitis, 10 microg/l prednisolone restored the smooth muscle responses to control values in preparations of trachea with intramural ganglia. After the prednisolone treatment, amplitude of the rat trachea muscle contraction in response to the nerve fiber electric stimulation did not differ statistically significantly from control and 0.1-10 microg/l prednisolone did not change the response value. The conclusion is made that prednisolone affected the diseased rats more efficiently than the healthy animals. The character of the glucocorticoid effect depends on the presence of intramural ganglia in the trachea wall.

[Results of applying molecular-and-genetic markers of chromosomal DNA in identification of unrecognizable remains of servicemen lost in military actions in the Northern Caucasus]. / Nekotorye rezul'taty ispol'zovaniia molekuliarno-geneticheskikh markerov khromosomnoi DNK dlia identifikatsii neopoznannykh ostankov voennosluzhashchikh, pogibshikh v khode boevykh deistvii na Severnom Kavkaze.

The results of applying molecular-and-genetic markers of chromosomal DNA in identifying unrecognizable remains of servicemen lost as dead in 1994-1996 and 1999-2001 military campaigns in the Northern Caucuses are summarized in the paper. Some of the specific features related with enzymatic amplification typing of DNA preparations sampled from degraded biological tissues of strongly deformed or decayed cadavers were analyzed. The typing results were analyzed by the AB0 system of degraded expertise samples in order to check the reliability of routine forensic-biological examinations as applicable to cadaver tissues with pronounced putrefactive changes. It was established that group-specific antigens of the AB0 system were correctly determined only in 67.5% of cases. False results were obtained in the other 32.5% of cases. Most of them were related with determination of groups 0(I) and A(II).

[Study of species specificity of the amelogenin system for genetic sex determination]. / Izuchenie vidovoi spetsifichnosti amelogeninovoi sistemy ustanovleniia geneticheskogo pola.

The properties of amelogenin amplification system and, in particular, of its species specificity, were studied. DNA preparations extracted from cattle (cow/bull), pig, ram and from poultry (hen), as well as from dog and cat, were used as a matrix for polymerase chain reaction (PCR) involving a standard scheme of enzymatic amplification of the amelogenin gene. It was demonstrated that, unlike for the human DNA, the amelogenin test couldn't be used for the DNA of examined animals as a sex-specific marker. However, there is a danger of a false determination of the male sex in the female origin samples during a forensic-experts' typing of the amelogenin gene, if there is an admixture of an animal DNA to a human DNA preparation. As for the biological samples of the animal origin, there is a possibility of a false-female sex determination. It can be attributed to an incorrectly selected standard of the molecular weight or to its remote location on the gel as well as to the use of an inaccurately calculated algorithm designed for determining the sizes of analyzed fragments.

[Analysis of genetic diversity of the mitochondrial DNA in the forensic medical expert identification of individuals]. / Analiz geneticheskogo raznoobraziia mitikhondrial'noi DNK v aspekte sudebno-ékspertnoi identifikatsii lichnosti.

Spectra of haplotype frequencies were studied for locuses of hypervariable segments 1 and 2 (HVS1 and HVS2), separately for each, and for linked segment HVS1-HVS2. The obtained data were used to determine the values and to evaluate comparatively the discriminating characteristics of the corresponding individualizing systems based on the typing of mtDNA. The system of typing, based on HVS2 (mv = 0.098), was found to possess the least discriminating potential; while the highest information rate is ensured in the analysis of HVS1 (m omega = 0.02) and in the joint analysis of HVS1 and HVS2 (mw = 0.007). The frequency rates of the key haplogroups were estimated within a random sampling of Russian citizens. A random population sampling of Russian citizens was shown not to differ essentially from an ethnically homogeneous population sampling of Russians selected with regard for a genetic diversity and for a spectrum of mitochondrial lines. The results point at the most rational algorithm of examinations in a forensic expert's analysis of mtDNA. The studied sampling can trigger the development of a referential data base designed for conducting, in the Russian Federation, the forensic-medical expert's examinations based on the mtDNA typing.

[Possible differentiation of mitotypes in mixed biological subjects based on phylogenetic analysis of mitochondrial DNA]. / O vozmozhnosti differentsirovaniia mitotipov v smeshannykh biologicheskikh ob''ektakh na osnove filogeneticheskogo analiza mitokhondrial'noi DNK.

A possibility was investigated to use the phylogenetic analysis of mitochondrial (mt) DNA to differentiate between the mito-types in mixed subjects in forensic medical examinations. The suggested methodological approach is based on the use of electrophoregram and on the comparison of such data with the information about phylogenic individual (mt) DNA lines.

[Utilization of complex technologies of molecular genetics for forensic expert identification of unidentified remains of victims of terrorist acts in Moscow in 1999]. / Kompleksnoe primenenie tekhnologii molekuliarno-geneticheskoi individualizatsii biologicheskikh ob''ektov dlia sudebno-ékspertnoi identifikatsii neopoznannykh ostankov zhertv terroristicheskikh aktov v Moskve v 1999g.

This paper presents the first experience gained in Russia in expert identification practice with complex utilization of technologies of molecular genetic individualization of biological objects for solving identification tasks requiring the maximum complete armory of mutually supplementing means. This is particularly important for forensic expert personality identification in cases with many deaths, when indirect identification has to be resorted to, consisting in the use of biological samples from relatives of the victims as the identifying objects. Methodological approaches and concepts of expert studies, used for identification of the remains of people who died in terroristic acts committed in Moscow in September, 1999, are discussed. The results of this study and methodological experience notably extend the potentialities of expert evaluation as regards forensic medical identification of victims of overall disasters, terroristic acts, and war conflicts.

[Allele polymorphism of molecular-genetic individualizing systems based on tetranucleotide tandem repeats LPL, vWA and TH01 among the population of Russia]. / Issledovanie allel'nogo polimorfizma molekuliarno-geneticheskikh individualiziruiushchikh sistem na osnove tetranukleotidnykh tandemnykh povtorov LPL, vWA, i TH01 sredi naseleniia Rossii.

Until recently, data bases on incidence rates of alleles STR (LPL, vWA and THO1 loci) have not been introduced in Russia. To obtain information on the above rates in Russia, a large scale research of distribution of LPL, vWA and Tho1 loci has been performed among a sample of 442 blood-unrelated persons from 57 regions of the Russian Federation to specify basic assessment characteristics of these loci and to raise efficiency of expert use of identifying systems on their base. Relevant parameters (allele frequencies) are presented for each polymorphic locus as well as basic population characteristics of polymorphism in the studied sample of RF population. These results can be used as key parameters for standard probability estimations in assessing the results of molecular-genetic identification.

[The experience of using the FTA-carriers for blood sample storage while conducting the molecular-genetic identification of unidentified killed persons--victims of military actions in the territory of the Chechen Republic]. / Opyt ispol'zovaniia FTA-nositelei dlia khraneniia odraztsov krovi pri provedenii molekuliarno-geneticheskikh identifikatsionnykh issledovanii neopoznannykh pogibshikh--zhertv voennykh deistvii na territorii Chechenskoi respubliki.

The activity of adapting and introducing the FTA-carriers for the storage of blood samples of relatives of missing service men for the purpose of using them in the consequent identification measures in respect to non-identified killed persons has been conducted in the Russian Federation within the framework of identification of service men killed in the Chechen territory during the 1994-1996 armed conflict. The article generalizes the experience of practical usage of FTA-cards as an element in the technological sequence of the molecular-genetic identification expertise. Methods related with application of FTA-technologies were tested; the efficiency of polymerized chain reaction was evaluated by using the matrix of immobilized DNA for hypo variable segments HVS-1 and HVS-2 as well as for the D-loop of mitochondrial DNA and for chromosomal STR-locuses D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820.

[Mitochondrial DNA typing--a new level for solving identification problems in forensic medical expert identification of unidentified remains of victims of terrorist acts in Moscow and the armed conflict in the Chechen Republic]. / Tipirovanie mitokhondrial'noi DNK--novyi uroven' resheniia identifikatsionnykh zadach pri sudebno-medintsinskoi ékspertize neopoznannykh ostankov zhertv terroristicheskikh aktov v Moskve i vooruzhennogo konflikta v Chechenskoi Respublike.

Two large-scale episodes described in this paper reflect the first in Russia use of molecular genetic matrilinear markers (analysis of polymorphism of sequences of amplified fragments of mitochondrial DNA hypervariable locuses) in solution of a complex identification problem: forensic medical identification of unidentified fragments of victims of explosions of houses in Moscow in September, 1999, and of soldiers dead in the war conflict in the Chechen Republic in 1994-1996. The results of this work and methodological experience gained in it essentially extend the potentialities of expert studies as regards forensic medical identification of victims of large scale disasters, terroristic acts, and war conflicts.

[Effect of histamine on duration of phases of the respiration cycle]. / Vliianie gistamina na dlitel'nost' faz dykhatel'nogo tsikla.

Histamine produced either a bronchodilation or a bronchoconstriction in rats. In a 0.01-1.0 mcg/ml concentration histamine augmented the contractions amplitude in electrical stimulation of the trachea, in a 10-100 mcg/ml concentration histamine enhances the muscle tone thus decreasing the induced contractions. The histamine effects seems to be connected with its prevailing influence on different structures of the airways depending on the concentration. Its high concentrations act directly on the smooth muscle whereas its lower concentration affects receptors signaling the functional modules of the metasympathetic nervous system within the intramural ganglia of the trachea.

[The electrophoretic properties of gels of high-resolution MS-4 agarose for separating DNA fragments in molecular genetic analysis]. / Issledovanie élektroforeticheskikh svoistv gelei vysokorazreshaiushchei agarozy MS-4 dlia razdeleniia fragmentov DNK v molekuliarno-geneticheskom analize.

The Hispanlab firm (Spain) offers a new series of Molecular Screen analytical agarose, including MS-4 agarose intended for separation of small fragments of DNA. This low-endosmotic agarose attracts attention as a probable alternative to well-known FMC Bio Products special agarose (USA) due to its useful properties. We characterized a gel-electrophoretic system based on MS-4 agarose (and its correspondence to its passport) and tested its fitness for standard molecular genetic expert investigations. The following parameters were assessed: mechanical properties of gel, its transparency, degree of background fluorescence upon ethidium bromide staining, resolving power, and position distortions in the electrophoregram. Important features of MS-4 agarose gels were detected, not typical of other agarose systems, which should be borne in mind during investigations with this agarose.