Natural male hybrid common shrews with a very long chromosomal multivalent at meiosis appear not to be completely sterile.

Among 36 known chromosomal hybrid zones of the common shrew Sorex araneus, the Moscow-Seliger hybrid zone is of special interest because inter-racial complex heterozygotes (F1 hybrids) produce the longest meiotic configuration, consisting of 11 chromosomes with monobrachial homology (undecavalent or chain-of-eleven: CXI). Different studies suggest that such a multivalent may negatively affect meiotic progression and in general should significantly reduce fertility of hybrids. In this work, by immunocytochemical and electron microscopy methods, we investigated for the first time chromosome synapsis, recombination and meiotic silencing in pachytene spermatocytes of natural inter-racial heterozygous shrew males carrying CXI configurations. Despite some abnormalities detected in spermatocytes, such as associations of chromosomes, stretched centromeres, and the absence of recombination nodules in some arms of the multivalent, a large number of morphologically normal spermatozoa were observed. Possible low stringency of pachytene checkpoints may mean that even very long meiotic configurations do not cause complete sterility of such complex inter-racial heterozygotes.

Fluctuating fine-scale spatial genetic structure in the common shrews (Eulipotyphla, Mammalia).

The spatial genetic structure is a topical issue in the studies of various aspects of ecology and evolution. Using the multilocus autocorrelation method with hypervariable microsatellite genetic markers, we investigated a fine-scale pattern of genetic structure in 5 local populations of the common shrew Sorex araneus separated by distances of 300-1000 m (the Moscow chromosomal race). Spatial genetic autocorrelation analysis based on 5 microsatellite loci (expected heterozygosity >0.79) with 90 alleles revealed a consistent pattern of significant positive genetic structure. By testing the autocorrelation at multiple scales from 25 to 500 m, we found that positive spatial genetic structure is detectable in distance classes of <500 m. The weaker spatial genetic structure positively correlated with a higher ratio of nonresident individuals to residents' activity (number of captures). In contrast to nonresident animals, the residents demonstrated prominent genetic structure. Genetic difference (FST ) between the populations was significant (0.016-0.051) and comparable with that between populations of different races analyzed previously (0.016-0.038). FST was not associated with geographic distance. These demographic patterns allowed us to propose a scheme of genetic-structure dynamics involving periodic appearance of more related local groups and renewal of allelic profiles from а common pool where the alleles are mixed. The scheme predicts fluctuating genetic structure and random similar differences among local populations.

High diversity of small insectivorous mammals on Qinghai-Tibet Plateau and first description of karyotype for four endemics of China.

Among seven species of the order Eulipotyphla (from southern Gansu and northern Sichuan Provinces, Central China) studied cytogenetically, karyotypes of one talpid species, Uropsilus aff. soricipes (2n = 36, NFa = 54), and three soricid species, Chodsigoa hypsibia (2n = 65, NFa = 66), Sorex cansulus (2n = 42, NFa = 64) and Sorex thibetanus (2n = 42, NFa = 60), are described cytogenetically for the first time. All four species are endemic to China with distribution ranges restricted to the Qinghai-Tibet Plateau and adjacent mountain ranges. The Ch. hypsibia karyotype consists of mostly acrocentric autosomes and one metacentric pair of autosomes; besides, a B chromosome was identified. No polymorphism was detected among karyotypes of other species, including shrews Sorex bedfordiae (2n = 26, NFa = 44), Anourosorex squamipes (2n = 48, NFa = 92) and Crocidura suaveolens (2n = 40, NFa = 44). The Chinese shrew mole U. aff. soricipes and three shrew species (S. bedfordiae, Ch. hypsibia and A. squamipes) represent autochthonous fauna of Central/Western China, whereas S. thibetanus, S. cansulus and C. suaveolens belong to phylogenetic groups occurring mostly to the north and west from China; therefore, they should be considered relatively recent colonisers. Thus, considering the relationships of the species within phylogenetic groups, our results on karyotypes are in good agreement with molecular genetic data.

RNA Aptamers for Theranostics of Glioblastoma of Human Brain.

Conventional approaches for studying and molecular typing of tumors include PCR, blotting, omics, immunocytochemistry, and immunohistochemistry. The last two methods are the most used, as they enable detecting both tumor protein markers and their localizations within the cells. In this study, we have investigated a possibility of using RNA aptamers, in particular, 2'-F-pyrimidyl-RNA aptamer ME07 (48 nucleotides long), specific to the receptor of epidermal growth factor (EGFR, ErbB1, Her1), as an alternative to monoclonal antibodies for aptacytochemistry and aptahistochemistry for human glioblastoma multiforme (GBM). A specificity of binding of FAM-ME07 to the receptor on the tumor cells has been demonstrated by flow cytometry; an apparent dissociation constant for the complex of aptamer - EGFR on the cell has been determined; a number of EGFR molecules has been semi-quantitatively estimated for the tumor cell lines having different amount of EGFR: A431 (106 copies per cell), U87 (104 copies per cell), MCF7 (103 copies per cell), and ROZH, primary GBM cell culture derived from patient (104 copies per cell). According to fluorescence microscopy, FAM-ME07 interacts directly with the receptors on A431 cells, followed by its internalization into the cytoplasm and translocation to the nucleolus; this finding opens a possibility of ME07 application as an escort aptamer for a delivery of therapeutic agents into tumor cells. FAM-ME07 efficiently stains sections of GBM clinical specimens, which enables an identification of EGFR-positive clones within a heterogeneous tumor; and providing a potential for further studying animal models of GBM.

Karyotypic and molecular evidence supports the endemic Tibetan hamsters as a separate divergent lineage of Cricetinae.

The genus status of Urocricetus was defined recently based on morphological and molecular data. Even though the amount of evidence for a separate phylogenetic position of this genus among Cricetinae continues to increase, there is still no consensus on its relationship to other groups. Here we give the first comprehensive description of the U. kamensis karyotype (2n = 30, NFa = 50) including results of comparative cytogenetic analysis and detailed examination of its phylogenetic position by means of numerous molecular markers. The molecular data strongly indicated that Urocricetus is a distant sister group to Phodopus. Comparative cytogenetic data showed significant reorganization of the U. kamensis karyotype compared to karyotypes of all other hamsters investigated earlier. The totality of findings undoubtedly means that Urocricetus belongs to a separate divergent lineage of Cricetinae.

Who are you, Griselda? A replacement name for a new genus of the Asiatic short-tailed shrews (Mammalia, Eulipotyphla, Soricidae): molecular and morphological analyses with the discussion of tribal affinities.

The first genetic study of the holotype of the Gansu short-tailed shrew, Blarinella griselda Thomas, 1912, is presented. The mitochondrial analysis demonstrated that the type specimen of B. griselda is close to several recently collected specimens from southern Gansu, northern Sichuan and Shaanxi, which are highly distinct from the two species of Asiatic short-tailed shrews of southern Sichuan, Yunnan, and Vietnam, B. quadraticauda and B. wardi. Our analysis of four nuclear genes supported the placement of B. griselda as sister to B. quadraticauda / B. wardi, with the level of divergence between these two clades corresponding to that among genera of Soricinae. A new generic name, Parablarinella, is proposed for the Gansu short-tailed shrew. Karyotypes of Parablarinella griselda (2n = 49, NFa = 50) and B. quadraticauda (2n = 49, NFa = 62) from southern Gansu are described. The tribal affinities of Blarinellini and Blarinini are discussed.

First cytogenetic analysis of lesser gymnures (Mammalia, Galericidae, Hylomys) from Vietnam.

Gymnures are an ancient group of small insectivorous mammals and are characterized by a controversial taxonomic status and the lack of a description of karyotypes for certain species. In this study, conventional cytogenetic techniques (Giemsa, CBG- and GTG-banding, Ag-NOR), CMA3-DAPI staining, and fluorescent in situ hybridization (FISH) with telomeric DNA probes were used to examine for the first time the karyotypes of lesser gymnures of group Hylomyssuillus Müller, 1840 from northern and southern Vietnam. All studied specimens had karyotypes with 2n=48, NFa=64. C-positive heterochromatic blocks existed in centromeric regions of 7 bi-armed autosomes and the submetacentric X chromosome. The Y chromosome is a C-positive and dot-like. The nucleolus organizer regions resided terminally on the short arms of 2 small bi-armed pairs. Positive signals at the telomeres of all chromosomes were revealed by FISH. CMA3-positive blocks were localized on the telomeric and pericentric regions of most bi-armed and acrocentric chromosomes. Despite the large genetic distances between Hylomys Müller, 1840, lesser gymnures from H.suillus-group from northern and southern Vietnam have similar karyotypic characteristics.

The Microtus voles: Resolving the phylogeny of one of the most speciose mammalian genera using genomics.

Sequential rapid radiations pose some of the greatest difficulties in phylogenetics, especially when analysing only a small number of genetic markers. Given that most of the speciation events occur in quick succession at various points in time, this creates particular challenges in determining phylogenetic relationships, i.e. branching order and divergence times. With the development of high throughput sequencing, thousands of markers can now readily be used to tackle these issues. Microtus is a speciose genus currently composed of 65 species that evolved over the last 2 million years. Although it is a well-studied group, there is still phylogenetic uncertainty at various divergence levels. Building upon previous studies that generally used small numbers of mitochondrial and/or nuclear loci, in this genomic-scale study we used both mitochondrial and nuclear data to study the rapid radiation within Microtus, using partial mitogenomes and genotyping-by-sequencing (GBS) on seven species representing five Microtus subgenera and the main biogeographic ranges where this group occurs. Both types of genome (mitochondrial and nuclear) generated similar tree topologies, with a basal split of the Nearctic (M. ochrogaster) and Holarctic (M. oeconomus) species, and then a subdivision of the five Palearctic species into two subgroups. These data support the occurrence of two European radiations, one North American radiation, and a later expansion of M. oeconomus from Asia to both Europe and North America. We further resolved the positioning of M. cabrerae as sister group of M. agrestis and refute the claim that M. cabrerae should be elevated to its own genus (Iberomys). Finally, the data support ongoing speciation events, especially within M. agrestis, with high levels of genetic divergence between the three Evolutionarily Significant Units (ESUs) previously identified. Similar high levels of divergence were also found among ESUs within M. oeconomus and M. arvalis.

Dual mechanism of chromatin remodeling in the common shrew sex trivalent (XY 1Y 2).

Here we focus on the XY1Y2 condition in male common shrew Sorex araneus Linnaeus, 1758, applying electron microscopy and immunocytochemistry for a comprehensive analysis of structure, synapsis and behaviour of the sex trivalent in pachytene spermatocytes. The pachytene sex trivalent consists of three distinct parts: short and long synaptic SC fragments (between the X and Y1 and between the X and Y2, respectively) and a long asynaptic region of the X in-between. Chromatin inactivation was revealed in the XY1 synaptic region, the asynaptic region of the X and a very small asynaptic part of the Y2. This inactive part of the sex trivalent, that we named the 'head', forms a typical sex body and is located at the periphery of the meiotic nucleus at mid pachytene. The second part or 'tail', a long region of synapsis between the X and Y2 chromosomes, is directed from the periphery into the nucleus. Based on the distribution patterns of four proteins involved in chromatin inactivation, we propose a model of meiotic silencing in shrew sex chromosomes. Thus, we conclude that pachytene sex chromosomes are structurally and functionally two different chromatin domains with specific nuclear topology: the peripheral inactivated 'true' sex chromosome regions (part of the X and the Y1) and more centrally located transcriptionally active autosomal segments (part of the X and the Y2).

Intrachromosomal Rearrangements in Rodents from the Perspective of Comparative Region-Specific Painting.

It has long been hypothesized that chromosomal rearrangements play a central role in different evolutionary processes, particularly in speciation and adaptation. Interchromosomal rearrangements have been extensively mapped using chromosome painting. However, intrachromosomal rearrangements have only been described using molecular cytogenetics in a limited number of mammals, including a few rodent species. This situation is unfortunate because intrachromosomal rearrangements are more abundant than interchromosomal rearrangements and probably contain essential phylogenomic information. Significant progress in the detection of intrachromosomal rearrangement is now possible, due to recent advances in molecular biology and bioinformatics. We investigated the level of intrachromosomal rearrangement in the Arvicolinae subfamily, a species-rich taxon characterized by very high rate of karyotype evolution. We made a set of region specific probes by microdissection for a single syntenic region represented by the p-arm of chromosome 1 of Alexandromys oeconomus, and hybridized the probes onto the chromosomes of four arvicolines (Microtus agrestis, Microtus arvalis, Myodes rutilus, and Dicrostonyx torquatus). These experiments allowed us to show the intrachromosomal rearrangements in the subfamily at a significantly higher level of resolution than previously described. We found a number of paracentric inversions in the karyotypes of M. agrestis and M. rutilus, as well as multiple inversions and a centromere shift in the karyotype of M. arvalis. We propose that during karyotype evolution, arvicolines underwent a significant number of complex intrachromosomal rearrangements that were not previously detected.

"European" race-specific metacentrics in East Siberian common shrews (Sorex araneus): a description of two new chromosomal races, Irkutsk and Zima.

Karyotype studies of common shrews in the vicinity of Lake Baikal (Irkutsk Region, Eastern Siberia) resulted in the description of two new chromosomal races of Sorex araneus Linnaeus, 1758 (Lypotyphla, Mammalia), additional to 5 races formerly found in Siberia. In the karyotypes of 12 specimens from 3 locations, the polymorphism of metacentric and acrocentric chromosomes of the Robertsonian type was recorded and two distinct groups of karyotypes interpreted as the chromosomal races were revealed. They are geographically distant and described under the racial names Irkutsk (Ir) and Zima (Zi). Karyotypes of both races were characterized by species-specific (the same for all 74 races known so far) metacentric autosomes af, bc, tu and jl, and the typical sex chromosome system - XX/XY1Y2. The race-specific arm chromosome combinations include three metacentrics and four acrocentrics in the Irkutsk race (gk, hi, nq, m, o, p, r) and four metacentrics and two acrocentrics in the Zima race (gm, hi, ko, nq, p, r). Within the races, individuals with polymorphic chromosomes were detected (g/m, k/o, n/q, p/r). The presence of the specific metacentric gk allowed us to include the Irkutsk race into the Siberian Karyotypic Group (SKG), distributed in surrounding regions. The Zima race karyotype contained two metacentrics, gm and ko, which have been never found in the Siberian part of the species range, but appear as the common feature of chromosomal races belonging to the West European Karyotypic Group (WEKG). Moreover, the metacentrics of that karyotype are almost identical to the Åkarp race (except the heterozygous pair p/r) locally found in the southern Sweden. One of two Siberian races described here for the first time, the Zima race, occurs in an area considerably distant from Europe and shares the common metacentrics (gm, hi, ko) with races included in WEKG. This fact may support a hypothesis of independent formation of identical arm chromosome combinations due to occurrence of the same centric fusions in different parts of the species range.

Phenotypic variation across chromosomal hybrid zones of the common shrew (Sorex araneus) indicates reduced gene flow.

Sorex araneus, the Common shrew, is a species with more than 70 karyotypic races, many of which form parapatric hybrid zones, making it a model for studying chromosomal speciation. Hybrids between races have reduced fitness, but microsatellite markers have demonstrated considerable gene flow between them, calling into question whether the chromosomal barriers actually do contribute to genetic divergence. We studied phenotypic clines across two hybrid zones with especially complex heterozygotes. Hybrids between the Novosibirsk and Tomsk races produce chains of nine and three chromosomes at meiosis, and hybrids between the Moscow and Seliger races produce chains of eleven. Our goal was to determine whether phenotypes show evidence of reduced gene flow at hybrid zones. We used maximum likelihood to fit tanh cline models to geometric shape data and found that phenotypic clines in skulls and mandibles across these zones had similar centers and widths as chromosomal clines. The amount of phenotypic differentiation across the zones is greater than expected if it were dissipating due to unrestricted gene flow given the amount of time since contact, but it is less than expected to have accumulated from drift during allopatric separation in glacial refugia. Only if heritability is very low, Ne very high, and the time spent in allopatry very short, will the differences we observe be large enough to match the expectation of drift. Our results therefore suggest that phenotypic differentiation has been lost through gene flow since post-glacial secondary contact, but not as quickly as would be expected if there was free gene flow across the hybrid zones. The chromosomal tension zones are confirmed to be partial barriers that prevent differentiated races from becoming phenotypically homogenous.

Synaptonemal complex analysis of interracial hybrids between the Moscow and Neroosa chromosomal races of the common shrew Sorex araneus showing regular formation of a complex meiotic configuration (ring-of-four).

Immunocytochemical and electron microscopic analysis of synaptonemal complexes (SCs) was carried out for the first time in homozygotes and complex Robertsonian heterozygotes (hybrids) of the common shrew, Sorex araneus Linnaeus, 1758, from a newly discovered hybrid zone between the Moscow and the Neroosa chromosomal races. These races differ in four monobrachial homologous metacentrics, and closed SC tetravalent is expected to be formed in meiosis of a hybrid. Indeed, such a multivalent was found at meiotic prophase I in hybrids. Interactions between multivalent and both autosomes and/or the sex chromosomes were observed. For the first time we have used immunocytochemical techniques to analyse asynapsis in Sorex araneus and show that the multivalent pairs in an orderly fashion with complete synapsis. Despite some signs of spermatocytes arrested in the meiotic prophase I, hybrids had large number of active sperm. Thus, Moscow - Neroosa hybrid males that form a ring-of-four meiotic configuration are most likely not sterile. Our results support previous demonstrations that monobrachial homology of metacentrics of the common shrew does not lead to complete reproductive isolation between parapatric chromosomal races of the species.

N'-(2-Hy-droxy-benzyl-idene)-2-(hy-droxy-imino)-propano-hydrazide.

The mol-ecule of the title compound, C(10)H(11)N(3)O(3), adopts an all-trans conformation and is approxomately planar, the largest deviation from the least-squares plane through all non-H atoms being 0.261 (1) Å. An intra-molecular O-H⋯N hydrogen bond occurs. In the crystal, the mol-ecules are packed into layers lying parallel to the ab plane by π-stacking inter-actions between the benzene ring of one molecule and the C-N bond of the oxime group of another molecule; the shortest inter-molecular C⋯C separation within the layer is 3.412 (1) Å. The layers are connected by O-H⋯O and N-H⋯O hydrogen bonds.

1-{3-[1-(Hydroxyimino)ethyl]-4-methyl-1H-pyrazol-5-yl}ethanone.

In the title compound, C(8)H(11)N(3)O(2), the oxime and the acetyl groups adopt a transoid conformation, while the pyrazole H atom is localized in the proximity of the acetyl group and is cis with respect to the acetyl O atom. In the crystal, dimers are formed as the result of hydrogen-bonding inter-actions involving the pyrazole NH group of one mol-ecule and the carbonyl O atom of another. The dimers are associated into sheets via O-H⋯N hydrogen bonds involving the oxime hydroxyl and the unprotonated pyrazole N atom, generating a macrocyclic motif with six mol-ecules.

Dimeric versus polymeric coordination in copper(II) cationic complexes with bis(chelating) oxime and amide ligands.

A series of copper(ii) complexes with earlier reported ligands N,N'-bis(2-hydroxyiminopropionyl)-1,2-diaminoethane (H(2)pen) and 1,3-diaminopropane (H(2)pap) and novel open chain oxime and amide ligands, N,N'-bis(2-hydroxyiminopropionyl)-1,5-diaminopentane (H(2)papt), 1,6-diaminohexane (H(2)pah) and 1,7-diaminoheptane (H(2)pahp), were prepared and characterized by a variety of spectroscopic methods and X-ray structure analysis. Although a pH-potentiometric study revealed only monomeric species in solution, formation of both binuclear dimeric and 1D-polymeric topologies was found in solid state. In all complexes ligands provide bis-bidentate coordination to Cu(2+) ions via the nitrogen atoms of the oxime groups and the oxygen atoms of the amide groups to form CuN(2)O(2) planar fragments. Compound [Cu(Hpap)(ClO(4))(H(2)O)](x) was isolated in both dimeric (x = 2) and polymeric (x = n) state, representing an example of supramolecular isomerism. All the complexes are additionally stabilized by short intramolecular hydrogen bonds =N-O-H...O-N= between cis-arranged oximato-groups. Stabilization is more effective in the case of dimeric complexes compared to 1D-polymeric chain, which appears to be the reason of preferable dimerization versus polymerization in the studied set of compounds. The effect of polymethylene linker length on the conformation of the dimers is described.

µ-Peroxido-bis-[acetonitrile-bis-(ethyl-enediamine)-cobalt(III)] tetrakis(per-chlorate).

The title compound, [Co(2)(O(2))(CH(3)CN)(2)(C(2)H(8)N(2))(4)](ClO(4))(4), consists of centrosymmetric binuclear cations and perchlorate anions. Two Co(III) atoms, which have a slightly distorted octa-hedral coordination, are connected through a peroxido bridge; the O-O distance is 1.476 (3) Å. Both acetonitrile ligands are situated in a trans position with respect to the O-O bridge. In the crystal, the complex cations are connected by N-H⋯O hydrogen bonds between ethyl-endiamine NH groups and O atoms from the perchlorate anions and peroxide O atoms.

2-Hydroxy-amino-2-oxoacetohydrazide.

In the title compound, C(2)H(5)N(3)O(3), the hydroxamic group adopts an anti orientation with respect to the hydrazide group. In the crystal, mol-ecules are connected by N-H⋯O and O-H⋯N hydrogen bonds into zigzag chains along the c axis.

Bis{2-hydroxy-imino-N'-[1-(2-pyrid-yl)ethyl-idene]propanohydrazidato}zinc(II) dihydrate.

The title compound, [Zn(C(10)H(11)N(4)O(2))(2)]·2H(2)O, was prepared by the reaction between Zn(CH(3)COO)(2)·2H(2)O and 2-hydroxy-imino-N'-[1-(2-pyrid-yl)ethyl-idene]propano-hydrazide (Hpop). The central Zn(II) atom has a distorted tetra-gonal-bipyramidal coordination geometry formed by two amide O atoms and four N atoms of two azomethine and two pyridine groups. In the crystal, complex mol-ecules form layers parallel to the crystallographic b direction. The layers are connected by O-H⋯N and O-H⋯O hydrogen bonds involving the solvent water mol-ecules.

µ-Oxalato-bis-[(2,2'-bipyridyl)-copper(II)] bis(perchlorate) dimethyl-formamide disolvate monohydrate.

The title compound, [Cu(2)(C(2)O(4))(C(10)H(8)N(2))(4)](ClO(4))(2)·2C(3)H(7)NO·H(2)O, contains doubly charged centrosymmetric dinuclear oxalato-bridged copper(II) complex cations, perchlorate anions, and DMF and water solvate mol-ecules. In the complex cation, the oxalate ligand is coordinated in a bis-bidentate bridging mode to the Cu atoms. Each Cu atom has a distorted tetra-gonal-bipyramidal environment, being coordinated by two N atoms of the two chelating bipy ligands and two O atoms of the doubly deprotonated oxalate anion. Pairs of perchlorate anions and water mol-ecules are linked into recta-ngles by O-H⋯O bonds in which the perchlorate O atoms act as acceptors and the water mol-ecules as donors. Methyl groups of the DMF solvent molecule are disordered over two sites with occupancies of 0.453 (7):0.547 (7), and the water molecule is half-occupied.