Synthesis of an analogue of the bisphosphonate drug Ibandronate for targeted drug-delivery therapeutic strategies

An analogue of the bisphosphonate drug Ibandronate was prepared and coupled via a cleavable ester function to a bromoacetyl linker with specific reactivity for thiol groups. This compound should find useful applications in therapeutic strategies aiming to deliver bisphosphonate drugs specifically to cancer cells making use of proteins as vectors. The specific delivery of bisphosphonates to cancer cells instead of bone, the usual site of accumulation of these cytotoxic drugs, could greatly widen their therapeutic applications.

De novo generation of a non-segmented negative strand RNA virus with a bicistronic gene

Reverse genetics has facilitated the use of non-segmented negative strand RNA viruses (NNSV) as vectors. Currently, heterologous gene expression necessitates insertion of extra-numeral transcription units (ENTUs), which may alter the NNSV polar transcription gradient and attenuate growth relative to wildtype (Wt). We hypothesized that rescuing recombinant Sendai Virus (rSeV) with a bicistronic gene might circumvent this attenuation but still allow heterologous open reading frame (ORF) expression. Therefore, we used a 9-nucleotide sequence previously described with internal ribosome entry site (IRES) activity, which, when constructed as several repeats, synergistically increased the level of expression of the second cistron [Chappell, S.A., Edelman, G.M., Mauro, V.P., 2000. A 9-nt segment of a cellular mRNA can function as an internal ribosome entry site (IRES) and when present in linked multiple copies greatly enhances IRES activity. Proc. Natl. Acad. Sci. U.S.A. 97, 1536-1541]. We inserted the Renilla luciferase (rLuc) ORF, preceded by 1, 3 or 7 IRES copies, downstream of the SeV N ORF in an infectious clone. Corresponding rSeVs were successfully rescued. Interestingly, bicistronic rSeVs grew as fast as or faster than Wt rSeV. Furthermore, SeV gene transcription downstream of the N/rLuc gene was either equivalent to, or slightly enhanced, compared to Wt rSeV. Importantly, all rSeV/rLuc viruses efficiently expressed rLuc. IRES repetition increased rLuc expression at a multiplicity of infection of 0.1, although without evidence of synergistic enhancement. In conclusion, our approach provides a novel way of insertion and expression of foreign genes in NNSVs. (C) 2008 Elsevier B.V. All rights reserved.

Macroalgal introductions by hull fouling on recreational vessels: Seaweeds and sailors

Macroalgal invasions in coastal areas have been a growing concern during the past decade. The present study aimed to assess the role of hull fouling on recreational yachts as a vector for macroalgal introductions. Questionnaire and hull surveys were carried out in marinas in France and Spain. The questionnaires revealed that the majority of yacht owners are aware of seaweed introductions, usually undertake short range journeys, dry dock their boat at least once a year, and use antifouling paints. The hull survey showed that many in-service yachts were completely free of macroalgae. When present, fouling assemblages consisted mainly of one to two macroalgal species. The most commonly found species was the tolerant green seaweed Ulva flexuosa. Most of the other species found are also cosmopolitan and opportunistic. A few nonnative and potentially invasive Ceramiales (Rhodophyta) were found occasionally on in-service yachts. On the basis of the information gathered during interviews of yacht owners in the surveyed area, these occurrences are likely to be uncommon. However they can pose a significant risk of primary or secondary introductions of alien macroalgal species, especially in the light of the increase in yachting activities. With large numbers of recreational yachts and relatively rare occurrences of nonnative species on hulls, comprehensive screening programs do not seem justified or practical. The risks of transferring nonnative species may, however, be minimized by encouraging the behaviors that prevent fouling on hulls and by taking action against neglected boats before they can act as vectors.

Family Histories and Rural Inheritance in Kenya

Inheritance systems and practices have a key role in people’s ability to exit poverty, or, conversely, plunging them further into it. As land is the major asset in low-income developing countries, how property is passed on and divided between future generations is a significant factor. This paper looks at inheritance through minimally-structured interviews with several generations of Kenyan families, seeking to explain that the how and why of poverty can be understood in the wider family context. It analyses their fortunes and misfortunes over a given time period in the context of property ownership rights. It also looks at the impact of education and the inheritance of cultural capital. When both fertility and survival are high, traditional patterns of land inheritance can lead to progression sub-division of land with long-term adverse implications for sustainability. While inheritance in Kenya is male dominated, the paper nonetheless examines the position of women in the chain as vectors of male property rights. The application of male-oriented customary law where inheritance is concerned, rather than the use of statutory legislation, was found to be the reality for the overwhelming majority of the participants in the study.

Applied aspects of Rhodococcus genetics

Eubacteria of the genus Rhodococcus are a diverse group of microorganisms commonly found in many environmental niches from soils to seawaters and as plant and animal pathogens. They exhibit a remarkable ability to degrade many organic compounds and their economic importance is becoming increasingly apparent. Although their genetic organisation is still far from understood, there have been many advances in recent years. Reviewed here is the current knowledge of rhodococci relating to gene transfer, recombination, plasmid replication and functions, cloning vectors and reporter genes, gene expression and its control, bacteriophages, insertion sequences and genomic rearrangements. Further fundamental studies of Rhodococcus genetics and the application of genetic techniques to the these bacteria will be needed for their continued biotechnological exploitation.

Speaker Recognition In Noisy Conditions With Limited Training Data

In this paper we present a novel method for performing speaker recognition with very limited training data and in the presence of background noise. Similarity-based speaker recognition is considered so that speaker models can be created with limited training speech data. The proposed similarity is a form of cosine similarity used as a distance measure between speech feature vectors. Each speech frame is modelled using subband features, and into this framework, multicondition training and optimal feature selection are introduced, making the system capable of performing speaker recognition in the presence of realistic, time-varying noise, which is unknown during training. Speaker identi?cation experiments were carried out using the SPIDRE database. The performance of the proposed new system for noise compensation is compared to that of an oracle model; the speaker identi?cation accuracy for clean speech by the new system trained with limited training data is compared to that of a GMM trained with several minutes of speech. Both comparisons have demonstrated the effectiveness of the new model. Finally, experiments were carried out to test the new model for speaker identi?cation given limited training data and with differing levels and types of realistic background noise. The results have demonstrated the robustness of the new system.

Identification of essential operons with a rhamnose-inducible promoter in Burkholderia cenocepacia

Scanning of bacterial genomes to identify essential genes is of biological interest, for understanding the basic functions required for life, and of practical interest, for the identification of novel targets for new antimicrobial therapies. In particular, the lack of efficacious antimicrobial treatments for infections caused by the Burkholderia cepacia complex is causing high morbidity and mortality of cystic fibrosis patients and of patients with nosocomial infections. Here, we present a method based on delivery of the tightly regulated rhamnose-inducible promoter P(rhaB) for identifying essential genes and operons in Burkholderia cenocepacia. We demonstrate that different levels of gene expression can be achieved by using two vectors that deliver P(rhaB) at two different distances from the site of insertion. One of these vectors places P(rhaB) at the site of transposon insertion, while the other incorporates the enhanced green fluorescent protein gene (e-gfp) downstream from P(rhaB). This system allows us to identify essential genes and operons in B. cenocepacia and provides a new tool for systematically identifying and functionally characterizing essential genes at the genomic level.

Novel macrocyclic and acyclic cationic lipids for gene transfer: Synthesis and in vitro evaluation

The synthesis and in vitro evaluation of four cationic lipid gene delivery vectors, characterized by acyclic or macrocyclic, and saturated or unsaturated hydrophobic regions, is described. The synthesis employed standard protocols, including ring-closing metathesis for macrocyclic lipid construction. All lipoplexes studied, formulated from plasmid DNA and a liposome composed of a synthesized lipid, 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (EPC), and either 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or cholesterol as co-lipid, exhibited plasmid DNA binding and protection from DNase I degradation, and concentration dependent cytotoxicity using Chinese hamster ovary-K1 cells. The transfection efficiency of formulations with cholesterol outperformed those with DOPE, and in many cases the EPC/cholesterol control, and formulations with a macrocyclic lipid (+/- 10:1) outperformed their acyclic counterparts (+/- 3:1).

Microneedle-mediated vaccine delivery:harnessing cutaneous immunobiology to improve efficacy

INTRODUCTION: Breaching the skin's stratum corneum barrier raises the possibility of the administration of vaccines, gene vectors, antibodies and even nanoparticles, all of which have at least their initial effect on populations of skin cells. AREAS COVERED: Intradermal vaccine delivery holds enormous potential for improved therapeutic outcomes for patients, particularly those in the developing world. Various vaccine-delivery strategies have been employed, which are discussed in this review. The importance of cutaneous immunobiology on the effect produced by microneedle-mediated intradermal vaccination is also discussed. EXPERT OPINION: Microneedle-mediated vaccines hold enormous potential for patient benefit. However, in order for microneedle vaccine strategies to fulfill their potential, the proportion of an immune response that is due to the local action of delivered vaccines on skin antigen-presenting cells, and what is due to a systemic effect from vaccines reaching the systemic circulation, must be determined. Moreover, industry will need to invest significantly in new equipment and instrumentation in order to mass-produce microneedle vaccines consistently. Finally, microneedles will need to demonstrate consistent dose delivery across patient groups and match this to reliable immune responses before they will replace tried-and-tested needle-and-syringe-based approaches.

Architecture and Construction Management at Queens University Belfast:Regional Cultural Centre

The Regional Cultural Centre in Letterkenny is a new 2000sqm arts center containing theatre, galleries, workshops and ancillary offices. The site is set back from the street, on high ground with good views. The form and envelope of the building was derived from geometrically connecting the site with the town’s two other main public buildings, the Cathedral (1901) and new Civic Offices (2002, also designed by MacGabhann Architects). This geometrical connection or vectors informed the geometry and shape of the building. This urban matrix of geometrically connecting three corner stones of society, namely the ecclesiastical headquarters, the administrative head quarters and the art centre helps to improve the town planning and urban design of the disparate and chaotic development that Letterkenny has become.
The large cantilever, which houses a 300sqm gallery, is aligned towards the Civic Offices, marks the entrance, and signifies a change of direction of the pedestrian route past the building, like a modern day obelisk.
The circulation routes and stairs internally provide views towards the civic offices and cathedral, thus reinforcing the connection between the three buildings and helps visitors make some sense of Letterkenny as an urban center. The main stairs and vertical circulation are contained behind the large glazed foyer, which is framed to be viewed externally like a proscenium stage, with visitors to the building passively acting their routes through the building.

A sequential algorithm for sparse support vector classifiers

Support vector machines (SVMs), though accurate, are not preferred in applications requiring high classification speed or when deployed in systems of limited computational resources, due to the large number of support vectors involved in the model. To overcome this problem we have devised a primal SVM method with the following properties: (1) it solves for the SVM representation without the need to invoke the representer theorem, (2) forward and backward selections are combined to approach the final globally optimal solution, and (3) a criterion is introduced for identification of support vectors leading to a much reduced support vector set. In addition to introducing this method the paper analyzes the complexity of the algorithm and presents test results on three public benchmark problems and a human activity recognition application. These applications demonstrate the effectiveness and efficiency of the proposed algorithm.


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On supercyclicity of operators from a supercyclic semigroup

We show that for every supercyclic strongly continuous operator
semigroup $\{T_t\}_{t\geq 0}$ acting on a complex $\F$-space, every
$T_t$ with $t>0$ is supercyclic. Moreover, the set of supercyclic
vectors of $T_t$ does not depend on the choice of $t>0$.

Design and test of a bit parallel 2nd order IIR filter structure

Test procedures for a pipelined bit-parallel IIR filter chip which maximally exploit its regularity are described. It is shown that small modifications to the basic architecture result in significant reductions in the number of test patterns required to test such chips. The methods used allow 100% fault coverage to be achieved using less than 1000 test vectors for a chip which has 12 bit data and coefficients.

Using sketch-map coordinates to analyze and bias molecular dynamics simulations

When examining complex problems, such as the folding of proteins, coarse grained descriptions of the system drive our investigation and help us to rationalize the results. Oftentimes collective variables (CVs), derived through some chemical intuition about the process of interest, serve this purpose. Because finding these CVs is the most difficult part of any investigation, we recently developed a dimensionality reduction algorithm, sketch-map, that can be used to build a low-dimensional map of a phase space of high-dimensionality. In this paper we discuss how these machine-generated CVs can be used to accelerate the exploration of phase space and to reconstruct free-energy landscapes. To do so, we develop a formalism in which high-dimensional configurations are no longer represented by low-dimensional position vectors. Instead, for each configuration we calculate a probability distribution, which has a domain that encompasses the entirety of the low-dimensional space. To construct a biasing potential, we exploit an analogy with metadynamics and use the trajectory to adaptively construct a repulsive, history-dependent bias from the distributions that correspond to the previously visited configurations. This potential forces the system to explore more of phase space by making it desirable to adopt configurations whose distributions do not overlap with the bias. We apply this algorithm to a small model protein and succeed in reproducing the free-energy surface that we obtain from a parallel tempering calculation.

Simplifying the representation of complex free-energy landscapes using sketch-map

A new scheme, sketch-map, for obtaining a low-dimensional representation of the region of phase space explored during an enhanced dynamics simulation is proposed. We show evidence, from an examination of the distribution of pairwise distances between frames, that some features of the free-energy surface are inherently high-dimensional. This makes dimensionality reduction problematic because the data does not satisfy the assumptions made in conventional manifold learning algorithms We therefore propose that when dimensionality reduction is performed on trajectory data one should think of the resultant embedding as a quickly sketched set of directions rather than a road map. In other words, the embedding tells one about the connectivity between states but does not provide the vectors that correspond to the slow degrees of freedom. This realization informs the development of sketch-map, which endeavors to reproduce the proximity information from the high-dimensionality description in a space of lower dimensionality even when a faithful embedding is not possible.