Multi-class, multi-residue determination of 132 veterinary drugs in milk by magnetic solid-phase extraction based on magnetic hypercrosslinked polystyrene prior to their determination by high-performance liquid chromatography-tandem mass spectrometry.

A quantitative multi-class multi-residue analytical method was developed for the determination of veterinary drugs in milk by high-performance liquid chromatography - tandem mass spectrometry (HPLC-MS/MS). A total of 132 veterinary drugs investigated belonged to almost 15 classes including sulfonamides, ß-lactams, tetracyclines, quinolones, macrolides, nitrofurans, nitroimidazoles, phenicols, lincosamides, pleuromutilins, macrocyclic lactones, quinoxaline antibiotics, benzimidazoles, anthelmintics, coccidiostats and some others. A magnetic solid-phase extraction procedure was developed using magnetic hypercrosslinked polystyrene (HCP/Fe3O4) for the sample preparation prior to HPLC-MS/MS without deproteinization step. The results indicated recoveries of 85-107% for 14 sulfonamides, 85-120% for 13 ß-lactams, 89-115% for 4 tetracyclines, 82-119% for 14 quinolones, 82-115% for 8 macrolides, 97-109% for 4 nitrofurans, 84-115% for 10 nitroimidazoles, 89-114% for 3 phenicols, 86-111% for 3 lincosamides, 97-102% for 2 pleuromutilins, 72-88% for 4 macrocyclic lactones, 87-104% for 4 quinoxaline antibiotics, 76-119% for 21 benzimidazoles, 79-115% for 12 anthelmintics, 81-118% for 12 coccidiostats and 75-119 % for 5 unclassified drugs, with relative standard deviations (RSDs) of less than 20%, and the LOQs ranged from 0.05 to 1 µg kg-1. This methodology was then applied to field-collected real milk samples and trace levels of some veterinary drugs were detected.

Luminescent Complexes of the Trinuclear Silver(I) and Copper(I) Pyrazolates Supported with Bis(diphenylphosphino)methane.

The first example of trinuclear copper(I) and silver(I) pyrazolates adducts with a tertiary diphosphine (Ph2PCH2PPh2) retaining trimeric [MPz]3 core is reported. Despite rather strong M-P bonding, the complexes are able to undergo the dissociation of one M-P bond leading to the "merry-go-round" movement of P atoms over the M3 triangle. The copper complex displays emission from 1MLCT and 3MLCT states. The triplet and singlet states are separated by a relatively small energy gap (1080 cm-1) that triggers the thermally activated delayed fluorescence (TADF) behavior and leads to the worthy quantum yield of 41% at 298 K. The silver complex in the solid state and frozen solution shows dual emission originating from the 1IL and 3MLCT states that is dictated by the much higher energy difference between the emissive singlet and triplet as well as by the essentially different nature of these states.

[Comparative results of preimplantation genetic screening by array comparative genomic hybridization and new-generation sequencing].

Aneuploidies as quantitative chromosome abnormalities are a main cause of failed development of morphologically normal embryos, implantation failures, and early reproductive losses. Preimplantation genetic screening (PGS) allows a preselection of embryos with a normal karyotype, thus increasing the implantation rate and reducing the frequency of early pregnancy loss after IVF. Modern PGS technologies are based on a genome-wide analysis of the embryo. The first pilot study in Russia was performed to assess the possibility of using semiconductor new-generation sequencing (NGS) as a PGS method. NGS data were collected for 38 biopsied embryos and compared with the data from array comparative genomic hybridization (array-CGH). The concordance between the NGS and array-CGH data was 94.8%. Two samples showed the karyotype 47,XXY by array-CGH and a normal karyotype by NGS. The discrepancies may be explained by loss of efficiency of array-CGH amplicon labeling.

Comparison of the results of preimplantation genetic screening obtained by a-CGH and NGS methods from the same embryos.

Chromosomal aneuploidies are known for being the main cause of abnormal development of embryos with normal morphology, their implantation failure and early reproductive losses in IVF treatments. Preimplantation genetic screening (PGS) allows selecting embryos with normal chromosomal content and increases IVF treatment efficiency due to higher implantation rates and less frequent early pregnancy losses. New technologies used for PGS allow making genome-wide analysis of the presence of all chromosomes in embryos. This article presents our study of evaluation of two techniques used for PGS: previously developed and used in our laboratory a-CGH assay based on Agilent technology and newly tested semi-conductive NGS technique (Torrent technology).

First cage-like pentanuclear Co(ii)-silsesquioxane.

A new pentanuclear "cylinder"-like cobalt(ii) phenylsilsesquioxane [(PhSiO1.5)10(CoO)5(NaOH)] exhibits a slow relaxation of the magnetization and a high catalytic activity and stereoselectivity in the oxidation of alkanes and alcohols.

Two pathways of proton transfer reaction to (triphos)Cu(η(1)-BH4) via a dihydrogen bond [triphos = 1,1,1-tris(diphenylphosphinomethyl)ethane].

The interaction of the η(1)-tetrahydroborate copper(i) complex (triphos)Cu(η(1)-BH4) () with proton donors [CF3CH2OH (TFE), (CF3)2CHOH (HFIP), (CF3)3COH (PFTB), PhOH, p-NO2C6H4OH (PNP), p-NO2C6H4N[double bond, length as m-dash]NC6H4OH (PNAP), CF3OH] was a subject of a combined IR spectroscopic and theoretical investigation. Spectral (Δν) and thermodynamic (ΔH) parameters of dihydrogen bond (DHB) formation were determined experimentally. The terminal hydride ligand (characterized by the basicity factor Ej(BH) = 0.87 ± 0.01) is found to be a site of proton transfer which begins with nucleophilic substitution of BH4(-) by the alcohol oxygen atom on the copper center (BH pathway). The activation barrier computed for (CF3)2CHOH in CH2Cl2 - ΔG = 20.6 kcal mol(-1) - is in good agreement with the experimental value (ΔG = 20.0 kcal mol(-1)). An abnormal dependence of the reaction rate on the proton donor strength found experimentally in dichloromethane is explained computationally on the basis of the variation of the structural and energetic details of this process with the proton donor strength. In the second reaction mechanism found (CuH pathway), DHB complexes with the initial ROH coordination to the bridging hydride lead to B-Hbr bond cleavage with BH3 elimination. "Copper assistance" via the CuO interaction is not involved. This mechanism can be evoked to explain the occurrence of proton transfer in coordinating solvents.

[Genetic determinants of resistance of hospital-associated strains of Klebsiella pneumoniae to ß-lactam antibiotics isolated in neonates].

According to the results of analysis of whole genome sequencing, the presence of genes having resistance to ß-lactam antibiotics in hospital-associated strains of Klebsiella pneumoniae was studied. The strains were isolated from neonatal intensive care units. The data obtained were compared with the results of antimicrobial susceptibility testing of isolated microorganisms. Among other strains resistant to cephalosporins, the dominance of genes of CTX-M-type extended-spectrum ß-lactamases was shown. It was revealed that one of eight strains phenotypically resistant and moderately resistant to carbapenems have the blaOXA-48 carbapenemase gene.

[Repetitive DNA sequences as an indicator of the level of genetic isolation in fish].

Although the functional role is still unknown for most types of nuclear noncoding repetitive sequences, some of them proved to provide adequate phylogenetic and taxonomic markers for studying the genetic relationships of organisms at the species and within-species levels. Several markers were used in this work. First, microsatellite markers were used to examine populations varying in the extent of genetic subdivision in marine and anadromous fish, including the Chilean jack mackerel Trachurus murphyi, anadromous brown trout Salmo trutta, and isolated and anadromous char populations. Locus polymorphism was proportional to the gene flow between populations in all cases. Second, satellite DNA was used to study the phylogenetic relationships within the genera Salmo, Oncorhynchus, Salvelinus, and Coregonus. Genetic distances agreed well with the taxonomic relationships based on morphological traits and various biochemical markers and correlated with the evolutionary ages estimated for the groups by other markers. Third, RAPD PCR with a set of 20-mer primers was performed to study the genus Coregonus and anadromous and isolated populations and species of the genus Salvelinus. The resulting phylogenetic trees may help to resolve some disputable taxonomic issues for the groups. A comparison showed that several RAPD-detected sequences contain conserved fragments of coding sequences and polymorphic repeats (minisatellites) from intergenic regions or introns. The finding point to a nonrandom nature of repetitive DNA divergence and may reflect the evolution of the fish groups examined. Heterochromatic satellite repeats were assumed to contribute to generating a reproductive barrier.

[Genetic structure of walleye pollock Theragra chalcogramma (Gadidae, Pisces) from Bering Sea and Sea of Okhotsk].

The genetic population structure in 4 spawning aggregations from the Bering Sea of walleye pollock (Theragra chalcogramma, Pallas), that occupies a central place in ecosystems of the North Pacific and is an important object of fisheries, was investigated with the ten microsatellite loci Tch5, Tch10, Tch11, Tch12, Tch14, Tch16, Tch17, Tch19, Tch20 and Tch22. The spatially distanced sample from Sea of Okhotsk was a referent group. The polymorphism of the markers used accounted for 100%, heterozygozity ranged from 41 to 95% in the different populations. It was shown that the aggregations of interest are in goodness-to-fit HWE at whole,but Sea of Okhotsk sample demonstrated the sex bias in the null-allele number in locus Tch16. Loci Tch10, Tch20 and Tch22 were of the most discriminate power. The genetic distances between all pairs of samples were typical for marine fishes. The sample from underwater Shirshov ridge region that is the natural geographic barrier for Olutor-Karagin and Koryak walleye pollock stocks is presumably of mixed genesis. As evident from scatter plots of F(ST) and spatial autocorrelation approach, data obtained (when Shirshov sample is eliminated) are under isolation by distance model on a broad spatial scale. The influence of abiotic factors on the population structure of the Bering Sea walleye pollock is supposed.

Genetic structure of the Salvelinus genus chars from reservoirs of the Kuril Islands.

Genetic resemblance of chars Salvelinus alpinus krasheninnikovi (Salvelinus malma krasheninnikovi) of 35 samples collected in five Kuril Islands--Shumshu, Paramushir, Onekotan, Iturup, and Kunashir--has been studied by the PCR-RAPD method. In the limits of each island, both resident isolates and anadromous forms give strictly supported clusters distinct from samples from the other islands. The samples from five islands form three superclusters: the first from Kunashir and Iturup Islands, the second from Paramushir and Onekotan Islands, and the third from Shumshu Island. The possible reasons for genetic similarity of resident and anadromous forms of Dolly Varden chars inhabiting reservoirs of a definite island are considered (the founder effect, homing, limited migration).

[Population genetic structure of the char species of the Northern Kuril Islands and the rank of the Dolly Varden Char in the system of the genus Salvelinus (Salmonidae: Teleostei)].

Analysis of the taxonomic position of most species and forms of the char (genus Salvelinus, Salmonidae: Teleostei) was made based on RAPD-PCR. The material was represented by samples from 29 populations from the Kuril Islands, coast of the Sea of Okhotsk, Kamchatka, Chukotka, Taymyr, Transbaikalia, the Kola Peninsula, Svalbard, Finland, and North America. It was shown that the genus Salvelinus splits into three well-justified clusters: (1) all the forms assigned to the Salvelinus alpinus--S. malma complex; (2) two samples of the White-Spotted Char from the Southern Kuril Islands and from Kamchatka; (3) two North American species, S. fontinalis and S. namaycush (samples of the North American species S. confluentis were absent from the collection). Analysis of the absolute values of genetic disctances of the S. alpinus--S. malma forms relative to S. leucomaenis, S. fontinalis, and S. namaycush revealed distances approaching the species rank between the following isolates: Frolikh Char, Mountain Char, Black Lake Char, Goggle-Eyed Char, and Neyva Char. Samples of Dolly Varden currently considered as "S. malma", do not constitute a separate cluster, falling within the group of the Arctic char S. alpinus. This conclusion is supported by the analysis of the results of three series of experiments by R. Phillips on ITS1 ribosome genes (Pleute et al., 1992; Phillips et al., 1995; Phillips et al., 1999). This indicates the infraspecific rank of malma within S. alpinus. Isolated populations of "Salvethymus svetovidovi" from the lake Elgygytgyn (Chukotka Peninsula) and of the char from the lake Chyornoye (Onekotan Island), recently described as S. gritzenkoi (Vasil'eva, Stygar, 2000), fell withing the S. alpinus--S. malma complex, the Onekotan char grouped together with another isolate from the same island. Comparison of genetic distances between the samples showed that the differences between the two isolated of Onekotan and migratory forms of the Kuril Islands are approximately equal, yet the homogeneity of the Chyornoye sample is higher than that of the other samples. The revealed 330-nucleotide diagnostic sequence of the Onekotan lake isolate showed identity of part of the fragment with a section of expressed DNA from the library of EST clones derived from the gills of Salmo salar, this possibly indicates the adaptive character of the evolution.

[Health status evaluation in children]. / Ob otsenke zdorov'ia detei.

A dependence of a health status on a condition of adaptation and of functional reserves of child's organism was studied. It was shown that the health status of contemporary schoolchildren is characterized, alongside with a higher prevalence of poly-organs' and morphological-and-functional deviations and chronic pathologies, by impaired rates and a lack of harmonic physical development as well as by a low level of reserve organism abilities. A variety of measures is suggested to promote the existing system of physical education.

Crown compounds for anions: sandwich and half-sandwich complexes of cyclic trimetric perfluoro-o-phenylenemercury with polyhedral closo-[B10H10]2- and closo-[B12H12]2- anions.

It has been shown by IR and NMR spectroscopy that cyclic trimeric perfluoro-o-phenylenemercury (o-C6F4-Hg)3 (1) is capable of binding closo-[B10H10]2- and closo-[B12H12]2- anions to form complexes [[(o-C6F4Hg)3](B10-H10)]2- (2), [[(o-C6F4Hg)3]2(B10H10)]2-(3), [[(o-C6F4Hg)3](B12H12)]2- (4), and [[(o-C6F4Hg)3]2(B12H12)]2- (5). According to IR data, the bonding of the [B10H10]2- and [B12H12]2- ions to the macrocycle in these complexes is accomplished through the formation of B-H-Hg bridges. Complexes 2, 3, and 5 have been isolated in analytically pure form and have been characterized by spectroscopic means. X-ray diffraction studies of 3 and 5 have revealed that these compounds have unusual sandwich structures, in which the polyhedral di-anion is located between the planes of two molecules of 1 and is bonded to each of them through two types of B-H-Hg bridges. One type is the simultaneous coordination of a B-H group to all three Hg atoms of the macrocycle. The other type is the coordination of a B-H group to a single Hg atom of the cycle. According to X-ray diffraction data, complex 2 has an analogous but half-sandwich structure. The obtained complexes 2-5 are quite stable; their stability constants in THF/acetone (1:1) at 20 degrees C have been determined as 1.0 x 10(2)Lmol(-1), 2.6 x 10(3)L(2)mol(2), 0.7 x 10(2)Lmol(-1), and 0.98 x 10(3)L(2)mol(-2), respectively.

[Whitefish: a new mechanism of reproductive isolation]. / Sigovye ryby: novyi mekhanizm reproduktivnoi izoliatsii.

Taxonoprint (a modification of DNA restrictase analysis) allows to distinguish sympatric species, that do not mate or produce hybrid offspring that are sterile or not viable. It is shown that taxonoprints of whitefish are very similar of identical. Sympatric whitefish are continuing to be separate despite they easily mate in experiments and in nature (up to 30% of individuals in nature are hybrids) and hybrids offspring have some features of heterosis. However it appears that hybrids of the second generation are not viable and can exist only because of back crossing with parents. In allows to keep a species independence in the process of gene exchange and to use heterosis of the first generation. Similar isolation mechanism is determined for other fish families (Acipensepidae, Clupeidae, Cyprinidae, Percidae) and some mammals (camels, sheep, bulls).