Magnetic properties of a cobalt(II) dimer of pseudo tetrahedral geometry [Co2{(CO)9Co3C---COO}5, C14H19N2H]

The crystalline ion pair [Co₂{OOC---CCo₃(CO)₉}₅, C₁₀H₆(N(CH₃)₂)₂H] (1) presents unusual magnetic properties. The X-band EPR spectrum of 1 at room temperature presents two unresolved bands at g=1.98 and 4.55. At a low temperature (20 K), the cluster of clusters 1 presents a complicated spectrum with an intense signal at 1700 G. The magnetic susceptibility of 1 was fit to a two spin S₁=S₂=3/2 Heisenberg model, with J=11.2 cm⁻¹ and a g value of 2.3. There is no field dependence of the magnetization, which suggests intramolecular coupling between the two tetrahedral centers of the cluster. Molecular orbital modeling indicates a sigma path of exchange between two topologically non-equivalent cobalt(II) centers.     

Ecologically safe method for improved feather wastes biodegradation

Twenty thermophilic actinomycetes were selected that were able to degrade feather wastes in high degree (87–91%). Increased proteolytic and lipolytic activity and exopolysaccharide production of the strains growing on feathers suggested their important role in the complex process of feathers biodegradation. Highest soluble protein content was determined in the fermented broths of the strains 3H, 8H, 4C, M4, and 27A. Based on data obtained, a mixed culture of three selected thermoactinomycete strains – 3H, 8H and M4, has been designed that considerably improved the feathers digestion process. The addition of feathers from 0.7 to 3% and pH values from 7.5 to 8.5 provided highest soluble protein content in the fermented broth of the mixed culture. It was shown that the hydrolysate obtained after 72 h growth of the mixed culture on feathers is rich in soluble proteins and amino acids including essential ones like lysine, threonine, leucine, isoleucine and valine, and rare ones like threonine, proline and serine. Therefore, the obtained value-added protein hydrolysate could be used for preparation of fertilizers or soil amendments, as well as protein source in animal feeding. To our knowledge, this is first report of using mixed culture of thermoactinomycete strains for improved feathers biodegradation. The proposed ecologically safe method is simple and economically viable thus applicable on industrial scale.     

Comparative sequence analyses reveal sites of ancestral chromosomal fusions in the Indian muntjac genome

Background

Indian muntjac (Muntiacus muntjak vaginalis) has an extreme mammalian karyotype, with only six and seven chromosomes in the female and male, respectively. Chinese muntjac (Muntiacus reevesi) has a more typical mammalian karyotype, with 46 chromosomes in both sexes. Despite this disparity, the two muntjac species are morphologically similar and can even interbreed to produce viable (albeit sterile) offspring. Previous studies have suggested that a series of telocentric chromosome fusion events involving telomeric and/or satellite repeats led to the extant Indian muntjac karyotype.

Results

We used a comparative mapping and sequencing approach to characterize the sites of ancestral chromosomal fusions in the Indian muntjac genome. Specifically, we screened an Indian muntjac bacterial artificial-chromosome library with a telomere repeat-specific probe. Isolated clones found by fluorescence in situ hybridization to map to interstitial regions on Indian muntjac chromosomes were further characterized, with a subset then subjected to shotgun sequencing. Subsequently, we isolated and sequenced overlapping clones extending from the ends of some of these initial clones; we also generated orthologous sequence from isolated Chinese muntjac clones. The generated Indian muntjac sequence has been analyzed for the juxtaposition of telomeric and satellite repeats and for synteny relationships relative to other mammalian genomes, including the Chinese muntjac.

Conclusions

The generated sequence data and comparative analyses provide a detailed genomic context for seven ancestral chromosome fusion sites in the Indian muntjac genome, which further supports the telocentric fusion model for the events leading to the unusual karyotypic differences among muntjac species.     

A regression-based analytic model for capacity planning of multi-tier applications

The multi-tier implementation has become the industry standard for developing scalable client-server enterprise applications. Since these applications are performance sensitive, effective models for dynamic resource provisioning and for delivering quality of service to these applications become critical. Workloads in such environments are characterized by client sessions of interdependent requests with changing transaction mix and load over time, making model adaptivity to the observed workload changes a critical requirement for model effectiveness. In this work, we apply a regression-based approximation of the CPU demand of client transactions on a given hardware. Then, we use this approximation in an analytic model of a simple network of queues, each queue representing a tier, and show the approximation’s effectiveness for modeling diverse workloads with a changing transaction mix over time. Using two case studies, we investigate factors that impact the efficiency and accuracy of the proposed performance prediction models. Experimental results show that this regression-based approach provides a simple and powerful solution for efficient capacity planning and resource provisioning of multi-tier applications under changing workload conditions.

A sulfated glucuronofucan containing both fucofuranose and fucopyranose residues from the brown alga Chordaria flagelliformis

A fucoidan fraction composed of l-fucose, sulfate, and d-glucuronic acid in a molar proportion of about 1:1:0.25 and small amount of acetyl groups was isolated from the brown alga Chordaria flagelliformis. Several modified polysaccharides were prepared from the native fucoidan using solvolytic desulfation, carboxyl reduction, and partial acid hydrolysis. Polysaccharide structures were elucidated by methylation analysis and 1D and 2D NMR spectroscopy. The fucoidan was shown to contain a backbone of 3-linked α-l-fucopyranose residues, about one-third of which are glycosylated at C-2 by α-d-glucopyranosyluronic acid residues. About half of the latter residues are glycosylated at C-4 by single α-l-fucofuranose residues or by disaccharides α-l-Fucf-(1→2)-α-l-Fucf-(1→. Fucofuranose residues are mono- and disulfated at different positions, whereas some additional sulfate groups occupy C-2 and C-4 of the backbone, the latter position being also partially acetylated.     

Single VS ribozyme molecules reveal dynamic and hierarchical folding toward catalysis

Non-coding RNAs of complex tertiary structure are involved in numerous aspects of the replication and processing of genetic information in many organisms; however, an understanding of the complex relationship between their structural dynamics and function is only slowly emerging. The Neurospora Varkud Satellite (VS) ribozyme provides a model system to address this relationship. First, it adopts a tertiary structure assembled from common elements, a kissing loop and two three-way junctions. Second, catalytic activity of the ribozyme is essential for replication of VS RNA in vivo and can be readily assayed in vitro. Here we exploit single molecule FRET to show that the VS ribozyme exhibits previously unobserved dynamic and heterogeneous hierarchical folding into an active structure. Readily reversible kissing loop formation combined with slow cleavage of the upstream substrate helix suggests a model whereby the structural dynamics of the VS ribozyme favor cleavage of the substrate downstream of the ribozyme core instead. This preference is expected to facilitate processing of the multimeric RNA replication intermediate into circular VS RNA, which is the predominant form observed in vivo.     

The Contribution of the Blood Glutamate Scavenging Activity of Pyruvate to its Neuroprotective Properties in a Rat Model of Closed Head Injury

The removal of excess glutamate from brain fluids after acute insults such as closed head injury (CHI) and stroke is expected to prevent excitotoxicity and the ensuing long lasting neurological deficits. Since blood glutamate scavenging accelerates the removal of excess glutamate from brain into blood and causes neuroprotection, we have evaluated here whether the neuroprotective properties of pyruvate could be partly accounted to its blood glutamate scavenging activity. The neurological outcome of rats after CHI improved significantly when treated with intravenous pyruvate (0.9 mmoles/100 g) but not with pyruvate administered together with glutamate. Pyruvate, at 5 μmole/100 g rat was neither protective not able to decrease blood glutamate but displayed the latter two properties when combined with 60 μg/100 g of glutamate-pyruvate transaminase. Since the neurological recovery from CHI was correlated with the decrease of blood glutamate levels, we conclude that pyruvate blood glutamate scavenging activity contributes to the spectrum of its neuroprotective mechanisms.     

Heparin affin regulatory peptide/pleiotrophin negatively affects diverse biological activities in C6 glioma cells

Heparin affin regulatory peptide (HARP) or pleiotrophin seems to be involved in the progression of several tumors of diverse origin. In this study, we tried to determine the role of HARP in rat C6 glioma cells by using an antisense strategy for inhibition of HARP expression. Decrease of the expression of endogenous HARP in C6 cells (AS-C6 cells) significantly increased proliferation, migration, and anchorage-independent growth of cells. Implantation of AS-C6 cells onto chicken embryo chorioallantoic membranes resulted in a significant increase of tumor-induced angiogenesis compared with that induced by non-transfected or C6 cells transfected with the plasmid alone (PC-C6 cells). In the same line, conditioned medium from AS-C6 cells significantly increased endothelial cell proliferation, migration, and tube formation in vitro compared with the effect of conditioned medium from C6 or PC-C6 cells. Interestingly, vascular endothelial growth factor (VEGF) induced C6 cell proliferation and migration, and SU1496, a selective inhibitor of VEGF receptor 2 (VEGFR2), blocked increased glioma cell growth, migration, and angiogenicity observed in AS-C6 cell cultures. The above results seem to be due to a direct interaction between HARP and VEGF in the culture medium of C6 and PC-C6 cells, while AS-C6 cells secreted comparable amounts of VEGF that do not interact with HARP. Collectively, these data suggest that HARP negatively affects diverse biological activities in C6 glioma cells, mainly due to binding of HARP to VEGF, which may sequester secreted VEGF from signalling through VEGFR2.     

MDA-MB-231 breast cancer cell viability,motility and matrix adhesion are regulated by a complex interplay of heparan sulfate,chondroitin−/dermatan sulfate and hyaluronan biosynthesis

Proteoglycans and glycosaminoglycans modulate numerous cellular processes relevant to tumour progression, including cell proliferation, cell-matrix interactions, cell motility and invasive growth. Among the glycosaminoglycans with a well-documented role in tumour progression are heparan sulphate, chondroitin/dermatan sulphate and hyaluronic acid/hyaluronan. While the mode of biosynthesis differs for sulphated glycosaminoglycans, which are synthesised in the ER and Golgi compartments, and hyaluronan, which is synthesized at the plasma membrane, these polysaccharides partially compete for common substrates. In this study, we employed a siRNA knockdown approach for heparan sulphate (EXT1) and heparan/chondroitin/dermatan sulphate-biosynthetic enzymes (β4GalT7) in the aggressive human breast cancer cell line MDA-MB-231 to study the impact on cell behaviour and hyaluronan biosynthesis. Knockdown of β4GalT7 expression resulted in a decrease in cell viability, motility and adhesion to fibronectin, while these parameters were unchanged in EXT1-silenced cells. Importantly, these changes were associated with a decreased expression of syndecan-1, decreased signalling response to HGF and an increase in the synthesis of hyaluronan, due to an upregulation of the hyaluronan synthases HAS2 and HAS3. Interestingly, EXT1-depleted cells showed a downregulation of the UDP-sugar transporter SLC35D1, whereas SLC35D2 was downregulated in β4GalT7-depleted cells, indicating an intricate regulatory network that connects all glycosaminoglycans synthesis. The results of our in vitro study suggest that a modulation of breast cancer cell behaviour via interference with heparan sulphate biosynthesis may result in a compensatory upregulation of hyaluronan biosynthesis. These findings have important implications for the development of glycosaminoglycan-targeted therapeutic approaches for malignant diseases.     

Reconstruction of radionuclide intakes for the residents of East Urals Radioactive Trace (1957–2011)

The East Urals Radioactive Trace (EURT) was formed after a chemical explosion in the radioactive waste-storage facility of the Mayak Production Association in 1957 (Southern Urals, Russia) and resulted in an activity dispersion of 7.4?×?10¹⁶ Bq into the atmosphere. Internal exposure due to ingestion of radionuclides with local foodstuffs was the main factor of public exposure at the EURT. The EURT cohort, combining residents of most contaminated settlements, was formed for epidemiological study at the Urals Research Center for Radiation Medicine, Russia (URCRM). For the purpose of improvement of radionuclide intake estimates for cohort members, the following data sets collected in URCRM were used: (1) Total β-activity and radiochemical measurements of ⁹⁰Sr in local foodstuffs over all of the period of interest (1958–2011; n?=?2200), which were used for relative ⁹⁰Sr intake estimations. (2) ⁹⁰Sr measurements in human bones and whole body (n?=?338); these data were used for average ⁹⁰Sr intake derivations using an age- and gender-dependent Sr-biokinetic model. Non-strontium radionuclide intakes were evaluated on the basis of ⁹⁰Sr intake data and the radionuclide composition of contaminated foodstuffs. Validation of radionuclide intakes during the first years after the accident was first carried out using measurements of the feces β-activity of EURT residents (n?=?148). The comparison of experimental and reconstructed values of feces β-activity shows good agreement. ⁹⁰Sr intakes for residents of settlements evacuated 7–14 days after the accident were also obtained from ⁹⁰Sr measurements in human bone and whole body. The results of radionuclide intake reconstruction will be used to estimate the internal doses for the members of the EURT cohort.