Quantitative single-molecule microscopy reveals that CENP-A(Cnp1) deposition occurs during G2 in fission yeast

The inheritance of the histone H3 variant CENP-A in nucleosomes at centromeres following DNA replication is mediated by an epigenetic mechanism. To understand the process of epigenetic inheritance, or propagation of histones and histone variants, as nucleosomes are disassembled and reassembled in living eukaryotic cells, we have explored the feasibility of exploiting photo-activated localization microscopy (PALM). PALM of single molecules in living cells has the potential to reveal new concepts in cell biology, providing insights into stochastic variation in cellular states. However, thus far, its use has been limited to studies in bacteria or to processes occurring near the surface of eukaryotic cells. With PALM, one literally observes and 'counts' individual molecules in cells one-by-one and this allows the recording of images with a resolution higher than that determined by the diffraction of light (the so-called super-resolution microscopy). Here, we investigate the use of different fluorophores and develop procedures to count the centromere-specific histone H3 variant CENP-A(Cnp1) with single-molecule sensitivity in fission yeast (Schizosaccharomyces pombe). The results obtained are validated by and compared with ChIP-seq analyses. Using this approach, CENP-A(Cnp1) levels at fission yeast (S. pombe) centromeres were followed as they change during the cell cycle. Our measurements show that CENP-A(Cnp1) is deposited solely during the G2 phase of the cell cycle

Non-Abelian Chern-Simons Vortices

We consider the bosonic sector of a N=2 supersymmetric Chern-Simons-Higgs theory in 2+1 dimensions. The gauge group is U(1)xU(N) and has N_f flavors of fundamental matter fields. The model supports non-Abelian (axially symmetric) vortices when N_f\geq N, which have internal (orientational) moduli. When N_f > N, the solutions acquire additional collective coordinates parameterizing their transverse size. We solve the BPS equations numerically and obtain local (N_f = N) and semi-local (N_f > N) string solutions.Comment: 12 pages, 3 figures. Shortened version, published in Phys. Lett.

Fv antibodies to aflatoxin B1 derived from a pre-immunized antibody phage display library system

The production and characterization of recombinant antibodies to aflatoxin B[SUB1] (AFB[SUB1]), a potent mycotoxin and carcinogen is described. The antibody fragments produced were then applied for use in a surface plasmon resonance-based biosensor (BIAcore), which measures biomolecular interactions in 'real-time'. Single chain Fv (scFv) antibodies were generated to aflatoxin B1 from an established phage display system, which incorporated a range of different plasmids for efficient scFv expression. The scFv's were used in the development of a competitive ELISA, and also for the development of surface plasmon resonance (SPR)-based inhibition immunoassays. They were found to be suitable for the detection of AFB[SUB1], in this format, with the assays being sensitive and reproducible

DySCo: Quantitating Associations of Membrane Proteins Using Two-Color Single-Molecule Tracking

We present a general method called dynamic single-molecule colocalization for quantitating the associations of single cell surface molecules labeled with distinct autofluorescent proteins. The chief advantages of the new quantitative approach are that, in addition to stable interactions, it is capable of measuring nonconstitutive associations, such as those induced by the cytoskeleton, and it is applicable to situations where the number of molecules is small

Connectivity mapping using a combined gene signature from multiple colorectal cancer datasets identified candidate drugs including existing chemotherapies

BACKGROUND: While the discovery of new drugs is a complex, lengthy and costly process, identifying new uses for existing drugs is a cost-effective approach to therapeutic discovery. Connectivity mapping integrates gene expression profiling with advanced algorithms to connect genes, diseases and small molecule compounds and has been applied in a large number of studies to identify potential drugs, particularly to facilitate drug repurposing. Colorectal cancer (CRC) is a commonly diagnosed cancer with high mortality rates, presenting a worldwide health problem. With the advancement of high throughput omics technologies, a number of large scale gene expression profiling studies have been conducted on CRCs, providing multiple datasets in gene expression data repositories. In this work, we systematically apply gene expression connectivity mapping to multiple CRC datasets to identify candidate therapeutics to this disease. RESULTS: We developed a robust method to compile a combined gene signature for colorectal cancer across multiple datasets. Connectivity mapping analysis with this signature of 148 genes identified 10 candidate compounds, including irinotecan and etoposide, which are chemotherapy drugs currently used to treat CRCs. These results indicate that we have discovered high quality connections between the CRC disease state and the candidate compounds, and that the gene signature we created may be used as a potential therapeutic target in treating the disease. The method we proposed is highly effective in generating quality gene signature through multiple datasets; the publication of the combined CRC gene signature and the list of candidate compounds from this work will benefit both cancer and systems biology research communities for further development and investigations

Bogomol'nyi Equations of Maxwell-Chern-Simons vortices from a generalized Abelian Higgs Model

We consider a generalization of the abelian Higgs model with a Chern-Simons term by modifying two terms of the usual Lagrangian. We multiply a dielectric function with the Maxwell kinetic energy term and incorporate nonminimal interaction by considering generalized covariant derivative. We show that for a particular choice of the dielectric function this model admits both topological as well as nontopological charged vortices satisfying Bogomol'nyi bound for which the magnetic flux, charge and angular momentum are not quantized. However the energy for the topolgical vortices is quantized and in each sector these topological vortex solutions are infinitely degenerate. In the nonrelativistic limit, this model admits static self-dual soliton solutions with nonzero finite energy configuration. For the whole class of dielectric function for which the nontopological vortices exists in the relativistic theory, the charge density satisfies the same Liouville equation in the nonrelativistic limit.Comment: 30 pages(4 figures not included), RevTeX, IP/BBSR/93-6

Recognizing Members of the Tournament Equilibrium Set is NP-hard

A recurring theme in the mathematical social sciences is how to select the "most desirable" elements given a binary dominance relation on a set of alternatives. Schwartz's tournament equilibrium set (TEQ) ranks among the most intriguing, but also among the most enigmatic, tournament solutions that have been proposed so far in this context. Due to its unwieldy recursive definition, little is known about TEQ. In particular, its monotonicity remains an open problem up to date. Yet, if TEQ were to satisfy monotonicity, it would be a very attractive tournament solution concept refining both the Banks set and Dutta's minimal covering set. We show that the problem of deciding whether a given alternative is contained in TEQ is NP-hard.Comment: 9 pages, 3 figure

Ceftriaxone-Resistant Salmonella Infection Acquired by a Child from Cattle

Background The emergence of resistance to antimicrobial agents within the salmonellae is a worldwide problem that has been associated with the use of antibiotics in livestock. Resistance to ceftriaxone and the fluoroquinolones, which are used to treat invasive salmonella infections, is rare in the United States. We analyzed the molecular characteristics of a ceftriaxone-resistant strain of Salmonella enterica serotype typhimurium isolated from a 12-year-old boy with fever, abdominal pain, and diarrhea. Methods We used pulsed-field gel electrophoresis and analysis of plasmids and β-lactamases to compare the ceftriaxone-resistant S. enterica serotype typhimurium from the child with four isolates of this strain obtained from cattle during a local outbreak of salmonellosis. Results The ceftriaxone-resistant isolate from the child was indistinguishable from one of the isolates from cattle, which was also resistant to ceftriaxone. Both ceftriaxone-resistant isolates were resistant to 13 antimicrobial agents; all but one of the resistance determinants were on a conjugative plasmid of 160 kb that encoded the functional group 1 β-lactamase CMY-2. Both ceftriaxone-resistant isolates were closely related to the three other salmonella isolates obtained from cattle, all of which were susceptible to ceftriaxone. Conclusions This study provides additional evidence that antibiotic-resistant strains of salmonella in the United States evolve primarily in livestock. Resistance to ceftriaxone, the drug of choice for invasive salmonella disease, is a public health concern, especially with respect to children, since fluoroquinolones, which can also be used to treat this disease, are not approved for use in children

The Next Big Match: Convergence, Competition and Sports Media Rights

Using examples from a number of different European countries, this article analyses the increasingly prominent position of traditional telecommunications companies, such as British Telecom (UK), Deutsche Telekom (Germany), France Telecom/Orange (France) and Telefonica (Spain), in the contemporary sports media rights market. The first part of the article examines the commercial strategies of telecommunications operators and highlights how their acquisition of sports rights has been driven by the need to ensure a competitive position within an increasingly converged communications market. The second part of the article then moves on to consider the regulation of the sports media rights market. Most significantly, this section emphasises the need for further regulatory intervention to ensure that increased competition for sports rights leads to improved services and lower prices for consumers, rather than merely endlessly spiralling fees for the exclusive ownership of premium rights that are then passed on to sports channel and/or broadband subscribers