Fibre-based devices for next generation photonics communication systems

The future broadband information network will undoubtedly integrate the mobility and flexibility of wireless access systems with the huge bandwidth capacity of photonics solutions to enable a communication system capable of handling the anticipated demand for interactive services. Towards wide coverage and low cost implementations of such broadband wireless photonics communication networks, various aspects of the enabling technologies are continuingly generating intense research interest. Among the core technologies, the optical generation and distribution of radio frequency signals over fibres, and the fibre optic signal processing of optical and radio frequency signals, have been the subjects for study in this thesis. Based on the intrinsic properties of single-mode optical fibres, and in conjunction with the concepts of optical fibre delay line filters and fibre Bragg gratings, a number of novel fibre-based devices, potentially suitable for applications in the future wireless photonics communication systems, have been realised. Special single-mode fibres, namely, the high birefringence (Hi-Bi) fibre and the Er/Yb doped fibre have been employed so as to exploit their merits to achieve practical and cost-effective all-fibre architectures. A number of fibre-based complex signal processors for optical and radio frequencies using novel Hi-Bi fibre delay line filter architectures have been illustrated. In particular, operations such as multichannel flattop bandpass filtering, simultaneous complementary outputs and bidirectional nonreciprocal wavelength interleaving, have been demonstrated. The proposed configurations featured greatly reduced environmental sensitivity typical of coherent fibre delay line filter schemes, reconfigurable transfer functions, negligible chromatic dispersions, and ease of implementation, not easily achievable based on other techniques. A number of unique fibre grating devices for signal filtering and fibre laser applications have been realised. The concept of the superimposed fibre Bragg gratings has been extended to non-uniform grating structures and into Hi-Bi fibres to achieve highly useful grating devices such as overwritten phase-shifted fibre grating structure and widely/narrowly spaced polarization-discriminating filters that are not limited by the intrinsic fibre properties. In terms of the-fibre-based optical millimetre wave transmitters, unique approaches based on fibre laser configurations have been proposed and demonstrated. The ability of the dual-mode distributed feedback (DFB) fibre lasers to generate high spectral purity, narrow linewidth heterodyne signals without complex feedback mechanisms has been illustrated. A novel co-located dual DFB fibre laser configuration, based on the proposed superimposed phase-shifted fibre grating structure, has been further realised with highly desired operation characteristics without the need for costly high frequency synthesizers and complex feedback controls. Lastly, a novel cavity mode condition monitoring and optimisation scheme for short length, linear-cavity fibre lasers has been proposed and achieved. Based on the concept and simplicity of the superimposed fibre laser cavities structure, in conjunction with feedback controls, enhanced output performances from the fibre lasers have been achieved. The importance of such cavity mode assessment and feedback control for optimised fibre laser output performance has been illustrated.

Data transmission by pulse compression

Pulse compression techniques originated in radar.The present work is concerned with the utilization of these techniques in general, and the linear FM (LFM) technique in particular, for comnunications. It introduces these techniques from an optimum communications viewpoint and outlines their capabilities.It also considers the candidacy of the class of LFM signals for digital data transmission and the LFM spectrum. Work related to the utilization of LFM signals for digital data transmission has been mostly experimental and mainly concerned with employing two rectangular LFM pulses (or chirps) with reversed slopes to convey the bits 1 and 0 in an incoherent node.No systematic theory for LFM signal design and system performance has been available. Accordingly, the present work establishes such a theory taking into account coherent and noncoherent single-link and multiplex signalling modes. Some new results concerning the slope-reversal chirp pair are obtained. The LFM technique combines the typical capabilities of pulse compression with a relative ease of implementation. However, these merits are often hampered by the difficulty of handling the LFM spectrum which cannot generally be expressed closed-form. The common practice is to obtain a plot of this spectrum with a digital computer for every single set of LFM pulse parameters.Moreover, reported work has been Justly confined to the spectrum of an ideally rectangular chirp pulse with no rise or fall times.Accordingly, the present work comprises a systerratic study of the LFM spectrum which takes the rise and fall time of the chirp pulse into account and can accommodate any LFM pulse with any parameters.It· formulates rather simple and accurate prediction criteria concerning the behaviour of this spectrum in the different frequency regions. These criteria would facilitate the handling of the LFM technique in theory and practice.

Recent patents and trends in orally disintegrating tablets

Orally disintegrating tablets (ODTs) offer many advantages over the conventional oral dosage forms in terms of convenience and ease of use. Over the last decade, substantial advances in the formulation of ODTs have been achieved in academia and industry that resulted in the emerging of a large number of patents. The aim of this review is to summarise the most recent patents in ODT formulations and highlight their motivations, inventive steps and significances in the development of ODT formulations. Five major techniques have been applied in manufacturing of ODTs, namely conventional tablet press, moulding, freeze drying, tablet loading and pulverization, with majority of the patents dedicated to the use of conventional tablet pressing. The patents have addressed various issues concerning the manufacturing of robust and practical ODT formulations by disclosing new manufacturing techniques, advantageous materials, and innovative formulation steps. However, future developments are required to reduce the cost and widening the application of the new manufacturing techniques, while simplifying and shortening the formulation steps will be crucial in the well established ones.

Tear protein interaction with hydrogel contact lenses

The design and synthesis of biomaterials covers a growing number of biomedical applications. The use of biomaterials in biological environment is associated with a number of problems, the most important of which is biocompatabUity. If the implanted biomaterial is not compatible with the environment, it will be rejected by the biological site. This may be manifested in many ways depending on the environment in which it is used. Adsorption of proteins takes place almost instantaneously when a biomaterial comes into contact with most biological fluids. The eye is a unique body site for the study of protein interactions with biomaterials, because of its ease of access and deceptive complexity of the tears. The use of contact lenses for either vision correction and cosmetic reasons or as a route for the controlled drug delivery, has significantly increased in recent years. It is relatively easy to introduce a contact lens Into the tear fluid and remove after a few minutes without surgery or trauma to the patient. A range of analytical techniques were used and developed to measure the proteins absorbed to some existing commercial contact lens materials and also to novel hydrogels synthesised within the research group. Analysis of the identity and quantity of proteins absorbed to biomaterials revealed the importance of many factors on the absorption process. The effect of biomaterial structure, protein nature in terms of size. shape and charge and pH of the environment on the absorption process were examined in order to determine the relative up-take of tear proteins. This study showed that both lysozyme and lactoferrin penetrate the lens matrix of ionic materials. Measurement of the mobility and activity of the protein deposited into the surface and within the matrix of ionic lens materials demonstrated that the mobility is pH dependent and, within the experimental errors, the biological activity of lysozyme remained unchanged after adsorption and desorption. The study on the effect of different monomers copolymerised with hydroxyethyl methacrylate (HEMA) on the protein up-take showed that monomers producing a positive charge on the copolymer can reduce the spoilation with lysozyme. The studies were extended to real cases in order to compare the patient dependent factors. The in-vivo studies showed that the spoilation is patient dependent as well as other factors. Studies on the extrinsic factors such as dye used in colour lenses showed that the addition of colourant affects protein absorption and, in one case, its effect is beneficial to the wearer as it reduces the quantity of the protein absorbed.

Effective knowledge transfer: a terminological perspective - Dismantling the jargon barrier to knowledge about computer security

The research is concerned with the terminological problems that computer users experience when they try to formulate their knowledge needs and attempt to access information contained in computer manuals or online help systems while building up their knowledge. This is the recognised but unresolved problem of communication between the specialist and the layman. The initial hypothesis was that computer users, through their knowledge of language, have some prior knowledge of the subdomain of computing they are trying to come to terms with, and that language can be a facilitating mechanism, or an obstacle, in the development of that knowledge. Related to this is the supposition that users have a conceptual apparatus based on both theoretical knowledge and experience of the world, and of several domains of special reference related to the environment in which they operate. The theoretical argument was developed by exploring the relationship between knowledge and language, and considering the efficacy of terms as agents of special subject knowledge representation. Having charted in a systematic way the territory of knowledge sources and types, we were able to establish that there are many aspects of knowledge which cannot be represented by terms. This submission is important, as it leads to the realisation that significant elements of knowledge are being disregarded in retrieval systems because they are normally expressed by language elements which do not enjoy the status of terms. Furthermore, we introduced the notion of `linguistic ease of retrieval' as a challenge to more conventional thinking which focuses on retrieval results.

Expert system design for cold form sections

The concept of an Expert System (ES) has been acknowledged as a very useful tool, but few studies have been carried out in its application to the design of cold rolled sections. This study involves primarily the use of an ES as a tool to improve the design process and to capture the draughtsman's knowledge. Its main purpose is to reduce substantially the time taken to produce a section drawing, thereby facilitating a speedy feedback to the customer. In order to communicate with a draughtsman, it is necessary to use sketches, symbolic representations and numerical data. This increases the complexity of programming an ES, as it is necessary to use a combination of languages so that decisions, calculations, graphical drawings and control of the system can be effected. A production system approach is used and a further step has been taken by introducing an Activator which is an autoexecute operation set up by the ES to operate an external program automatically. To speed up the absorption of new knowledge into the knowledge base, a new Learning System has been constructed. In addition to developing the ES, other software has been written to assist the design process. The section properties software has been introduced to improve the speed and consistency of calculating the section properties. A method of selecting or comparing the most appropriate section for a given specification is also implemented. Simple loading facilities have been introduced to guide the designer as to the loading capacity of the section. This research has concluded that the application of an ES is beneficial and with the activator approach, automated designing can be achieved. On average a complex drawing can be displayed on the screen in about 100 seconds, where over 95% of the initial section design time for repetitive or similar profile can be saved.

Melt processable biomaterials for degradable surgical fixation devices

There are currently few biomaterials which combine controlled degradation rates with ease of melt processability. There are however, many applications ranging from surgical fixation devices to drug delivery systems which require such combination properties. The work in this thesis is an attempt to increase the availability of such materials. Polyhydroxybutyrate-polyhydroxyvalerate copolymers are a new class of potentially biodegradable materials, although little quantitative data relating to their in vitro and in vivo degradation behaviour exists. The hydrolytic degradation of these copolymers has been examined in vitro under conditions ranging from `physiological' to extremes of pH and elevated temperature. Progress of the degradation process was monitored by weight loss and water uptake measurement, x-ray diffractometry, optical and electron microscopy, together with changes in molecular weight by gel permeation chromatography. The extent to which the degradation mechanism could be modified by forming blends with polysaccharides and polycaprolactone was also investigated. Influence of the valerate content, molecular weight, crystallinity, together with the physical form of the sample, the pH and the temperature of the aqueous medium on the hydrolytic degradation was investigated. Its progress was characterised by an initial increase in the wet weight, with concurrent decrease in the dry weight as the amorphous regions of the polymer are eroded, thereby producing an increase in matrix porosity. With the polysaccharide blends, this initial rate is dramatically affected, and erosion of the polysaccharide from the matrix markedly increases the internal porosity which leads to the eventual collapse of the matrix, a process which occurs, but less rapidly, in the degradation of the unblended polyhydroxybutyrate-polyhydroxyvalerate copolymers. Surface energy measurement and goniophotometry proved potentially useful in monitoring the early stages of the degradation, where surface rather than bulk processes predominate and are characterised by little weight loss.

Titanium phosphate glass microspheres for bone tissue engineering

We have demonstrated the successful production of titanium phosphate glass microspheres in the size range of ~10-200 µm using an inexpensive, efficient, easily scalable process and assessed their use in bone tissue engineering applications. Glasses of the following compositions were prepared by melt-quench techniques: 0.5P2O5-0.4CaO-(0.1 - x)Na2O-xTiO2, where x = 0.03, 0.05 and 0.07 mol fraction (denoted as Ti3, Ti5 and Ti7 respectively). Several characterization studies such as differential thermal analysis, degradation (performed using a novel time lapse imaging technique) and pH and ion release measurements revealed significant densification of the glass structure with increased incorporation of TiO2 in the glass from 3 to 5 mol.%, although further TiO2 incorporation up to 7 mol.% did not affect the glass structure to the same extent. Cell culture studies performed using MG63 cells over a 7-day period clearly showed the ability of the microspheres to provide a stable surface for cell attachment, growth and proliferation. Taken together, the results confirm that 5 mol.% TiO2 glass microspheres, on account of their relative ease of preparation and favourable biocompatibility, are worthy candidates for use as substrate materials in bone tissue engineering applications.

Amino acids in oral drug delivery:salts, ion-pairs and transcriptomics

Oral drug delivery is considered the most popular route of delivery because of the ease of administration, availability of a wide range of dosage forms and the large surface area for drug absorption via the intestinal membrane. However, besides the unfavourable biopharmaceutical properties of the therapeutic agents, efflux transporters such as Pglycoprotein (P-gp) and multiple resistance proteins (MRP) decrease the overall drug uptake by extruding the drug from the cells. Although, prodrugs have been investigated to improve drug partitioning by masking the polar groups covalently with pre-moieties promoting increased uptake, they present significant challenges including reduced solubility and increased toxicity. The current work investigates the use of amino acids as ion-pairs for three model drugs: indomethacin (weak acid), trimethoprim (weak base) and ciprofloxacin (zwitter ion) in an attempt to improve both solubility and uptake. Solubility was studied by salt formation while creating new routes for uptake across the membranes via amino acids transporter proteins or dipeptidyl transporters was the rationale to enhance absorption. New salts were prepared for the model drugs and the oppositely charged amino acids by freeze drying and they were characterised using FTIR, 1HNMR, DSC, SEM, pH solubility profile, solubility and dissolution. Permeability profiles were assessed using an in vitro cell based method; Caco-2 cells and the genetic changes occurring across the transporter genes and various pathways involved in the cellular activities were studied using DNA microarrays. Solubility data showed a significant increase in drug solubility upon preparing the new salts with the oppositely charged counter ions (ciprofloxacin glutamate salt exhibiting 2.9x103 fold enhancement when compared to the free drug). Moreover, permeability studies showed a 3 fold increase in trimethoprim and indomethacin permeabilities upon ion-pairing with amino acids and more than 10 fold when the zwitter ionic drug was paired with glutamic acid. Microarray data revealed that trimethoprim was absorbed actively via OCTN1 transporters while MRP7 is the main transporter gene that mediates its efflux. The absorption of trimethoprim from trimethoprim glutamic acid ion-paired formulations was affected by the ratio of glutamic acid in the formulation which was inversely proportional to the degree of expression of OCTN1. Interestingly, ciprofloxacin glutamic acid ion-pairs were found to decrease the up-regulation of ciprofloxacin efflux proteins (P-gp and MRP4) and over-express two solute carrier transporters; (PEPT2 and SLCO1A2) suggesting that a high aqueous binding constant (K11aq) enables the ion-paired formulations to be absorbed as one entity. In conclusion, formation of ion-pairs with amino acids can influence in a positive way solubility, transfer and gene expression effects of drugs.

Enabling effective user interactions in content-based image retrieval

This paper presents an interactive content-based image retrieval framework—uInteract, for delivering a novel four-factor user interaction model visually. The four-factor user interaction model is an interactive relevance feedback mechanism that we proposed, aiming to improve the interaction between users and the CBIR system and in turn users overall search experience. In this paper, we present how the framework is developed to deliver the four-factor user interaction model, and how the visual interface is designed to support user interaction activities. From our preliminary user evaluation result on the ease of use and usefulness of the proposed framework, we have learnt what the users like about the framework and the aspects we could improve in future studies. Whilst the framework is developed for our research purposes, we believe the functionalities could be adapted to any content-based image search framework.

Silica-clad neodymium-doped lanthanum phosphate fibers and fiber lasers

We have fabricated a neodymium-doped phosphate glass fiber with a silica cladding and used it to form a fiber laser. Phosphate and silicate glasses have considerably different glass transition temperatures and softening points making it hard to draw a fiber from these two glasses. A bulk phosphate glass of composition (Nd2O3)(0.011)(La2O3)(0.259)(P2O5)(0.725)(Al2O3)(0.005) was prepared and the resultant material was transparent, free from bubbles and visibly homogeneous. The bulk phosphate glass was drawn to a fiber while being jacketed with silica and the resultant structure was of good optical quality, free from air bubbles and major defects. The attenuation at a wavelength of 1.06 mu m was 0.05 dB/cm and the refractive index of the core and cladding at the pump wavelength of 488 nm was 1.56 and 1.46, respectively. The fibers were mechanically strong enough to allow for ease of handling and could be spliced to conventional silica fiber. The fibers were used to demonstrate lasing at the F-4(3/2) - I-4(11/2) (1.06 mu m) transition. Our work demonstrates the potential to form silica clad optical fibers with phosphate cores doped with very high levels of rare-earth ions (27-mol % rare-earth oxide).

Editorial

The American Academy of Optometry (AAO) had their annual meeting in San Diego in December 2005 and the BCLA and CLAE were well represented there. The BCLA does have a reasonable number of non-UK based members and hopefully in the future will attract more. This will certainly be beneficial to the society as a whole and may draw more delegates to the BCLA annual conference. To increase awareness of the BCLA at the AAO a special evening seminar was arranged where BCLA president Dr. James Wolffsohn gave his presidential address. Dr. Wolffsohn has given the presidential address in the UK, Ireland, Hong Kong and Japan – making it the most travelled presidential address for the BCLA to date. Aside from the BCLA activity at the AAO there were numerous lectures of interest to all, truly a “something for everyone” meeting. All the sessions were multi-track (often up to 10 things occurring at the same time) and the biggest dilemma was often deciding what to attend and more importantly what you will miss! Nearly 200 new AAO Fellows were inducted at the Gala Dinner from many countries including 3 new fellows from the UK (this year they all just happened to be from Aston University!). It is certainly one of the highlights of the AAO to see fellows from different schools of training from around the world fulfilling the same criteria and being duly rewarded for their commitment to the profession. BCLA members will be aware that 2006 sees the introduction of the new fellowship scheme of the BCLA and by the time you read this the first set of fellowship examinations will have taken place. For more details of the FBCLA scheme see the BCLA web site http://www.bcla.org.uk. Since many of CLAE's editorial panel were at the AAO an informal meeting and dinner was arranged for them where ideas were exchanged about the future of the journal. It is envisaged that the panel will meet twice a year – the next meeting will be at the BCLA conference. The biggest excitement by far was the fact that CLAE is now Medline/PubMed indexed. You may ask why is this significant to CLAE? PubMed is the free web-based service from the US National Library of Medicine. It holds over 15 million biomedical citations and abstracts from the Medline database. Medline is the largest component of PubMed and covers over 4800 journals published in more than 70 countries. The impact of this is that CLAE is starting to attract more submissions as researchers and authors are not worried that their work will not be hidden from other colleagues in the field but rather the work is available to view on the World Wide Web. CLAE is one of a very small number of contact lens journals that is indexed this way. Amongst the other CL journals listed you will note that the International Contact Lens Clinic has now merged with CLAE and the journal CLAO has been renamed Eye and Contact Lenses – making the list of indexed CL journals even smaller than it appears. The on-line submission and reviewing system introduced in 2005 has also made it easier for authors to submit their work and easier for reviewers to check the content. This ease of use has lead to quicker times from submission to publication. Looking back at the articles published in CLAE in 2005 reveals some interesting facts. The majority of the material still tends to be from UK groups related to the field of Optometry, although we hope that in the future we will attract more work from non-UK groups and also from non-Optometric areas such as refractive surgery or anterior eye pathology. Interestingly in 2005 the most downloaded article from CLAE was “Wavefront technology: Past, present and future” by Professor W. Neil Charman, who was also the recipient of the Charles F. Prentice award at the AAO – one of the highest awards honours that the AAO can bestow. Professor Charman was also the keynote speaker at the BCLA's first Pioneer's Day meeting in 2004. In 2006, readers of CLAE will notice more changes, firstly we are moving to 5 issues per year. It is hoped that in the future, depending on increased submissions, a move to 6 issues may be feasible. Secondly, CLAE will aim to have one article per issue that carries CL CET points. You will see in this issue there is an article from Professor Mark Wilcox (who was a keynote speaker at the BCLA conference in 2005). In future articles that carry CET points will be either reviews from BCLA conference keynote speakers, members of the editorial panel or material from other invited persons that will be of interest to the readership of CLAE. Finally, in 2006, you will notice a change to the Editorial Panel, some of the distinguished panel felt that it was good time to step down and new members have been invited to join the remaining panel. The panel represent some of the most eminent names in the fields of contact lenses and/or anterior eye and have varying backgrounds and interests from many of the prominent institutions around the world. One of the tasks that the Editorial Panel undertake is to seek out possible submissions to the journal, either from conferences they attend (posters and papers that they will see and hear) and from their own research teams. However, on behalf of CLAE I would like to extend that invitation to seek original articles to all readers – if you hear a talk and think it could make a suitable publication to CLAE please ask the presenters to submit the work via the on-line submission system. If you found the work interesting then the chances are so will others. CLAE invites submissions that are original research, full length articles, short case reports, full review articles, technical reports and letters to the editor. The on-line submission web page is http://www.ees.elsevier.com/clae/.

XML Editors Overview and the Challenge to Build XML-Oriented Editor for Mediaeval Manuscript Descriptions

* The research presented here is partially supported by the project KT-DigiCult-Bg (FP6) and by the ICT Agency in Bulgaria.

Editorial overview:new protein production tools for structural biology

Full text: The idea of producing proteins from recombinant DNA hatched almost half a century ago. In his PhD thesis, Peter Lobban foresaw the prospect of inserting foreign DNA (from any source, including mammalian cells) into the genome of a λ phage in order to detect and recover protein products from Escherichia coli [ 1 and 2]. Only a few years later, in 1977, Herbert Boyer and his colleagues succeeded in the first ever expression of a peptide-coding gene in E. coli — they produced recombinant somatostatin [ 3] followed shortly after by human insulin. The field has advanced enormously since those early days and today recombinant proteins have become indispensable in advancing research and development in all fields of the life sciences. Structural biology, in particular, has benefitted tremendously from recombinant protein biotechnology, and an overwhelming proportion of the entries in the Protein Data Bank (PDB) are based on heterologously expressed proteins. Nonetheless, synthesizing, purifying and stabilizing recombinant proteins can still be thoroughly challenging. For example, the soluble proteome is organized to a large part into multicomponent complexes (in humans often comprising ten or more subunits), posing critical challenges for recombinant production. A third of all proteins in cells are located in the membrane, and pose special challenges that require a more bespoke approach. Recent advances may now mean that even these most recalcitrant of proteins could become tenable structural biology targets on a more routine basis. In this special issue, we examine progress in key areas that suggests this is indeed the case. Our first contribution examines the importance of understanding quality control in the host cell during recombinant protein production, and pays particular attention to the synthesis of recombinant membrane proteins. A major challenge faced by any host cell factory is the balance it must strike between its own requirements for growth and the fact that its cellular machinery has essentially been hijacked by an expression construct. In this context, Bill and von der Haar examine emerging insights into the role of the dependent pathways of translation and protein folding in defining high-yielding recombinant membrane protein production experiments for the common prokaryotic and eukaryotic expression hosts. Rather than acting as isolated entities, many membrane proteins form complexes to carry out their functions. To understand their biological mechanisms, it is essential to study the molecular structure of the intact membrane protein assemblies. Recombinant production of membrane protein complexes is still a formidable, at times insurmountable, challenge. In these cases, extraction from natural sources is the only option to prepare samples for structural and functional studies. Zorman and co-workers, in our second contribution, provide an overview of recent advances in the production of multi-subunit membrane protein complexes and highlight recent achievements in membrane protein structural research brought about by state-of-the-art near-atomic resolution cryo-electron microscopy techniques. E. coli has been the dominant host cell for recombinant protein production. Nonetheless, eukaryotic expression systems, including yeasts, insect cells and mammalian cells, are increasingly gaining prominence in the field. The yeast species Pichia pastoris, is a well-established recombinant expression system for a number of applications, including the production of a range of different membrane proteins. Byrne reviews high-resolution structures that have been determined using this methylotroph as an expression host. Although it is not yet clear why P. pastoris is suited to producing such a wide range of membrane proteins, its ease of use and the availability of diverse tools that can be readily implemented in standard bioscience laboratories mean that it is likely to become an increasingly popular option in structural biology pipelines. The contribution by Columbus concludes the membrane protein section of this volume. In her overview of post-expression strategies, Columbus surveys the four most common biochemical approaches for the structural investigation of membrane proteins. Limited proteolysis has successfully aided structure determination of membrane proteins in many cases. Deglycosylation of membrane proteins following production and purification analysis has also facilitated membrane protein structure analysis. Moreover, chemical modifications, such as lysine methylation and cysteine alkylation, have proven their worth to facilitate crystallization of membrane proteins, as well as NMR investigations of membrane protein conformational sampling. Together these approaches have greatly facilitated the structure determination of more than 40 membrane proteins to date. It may be an advantage to produce a target protein in mammalian cells, especially if authentic post-translational modifications such as glycosylation are required for proper activity. Chinese Hamster Ovary (CHO) cells and Human Embryonic Kidney (HEK) 293 cell lines have emerged as excellent hosts for heterologous production. The generation of stable cell-lines is often an aspiration for synthesizing proteins expressed in mammalian cells, in particular if high volumetric yields are to be achieved. In his report, Buessow surveys recent structures of proteins produced using stable mammalian cells and summarizes both well-established and novel approaches to facilitate stable cell-line generation for structural biology applications. The ambition of many biologists is to observe a protein's structure in the native environment of the cell itself. Until recently, this seemed to be more of a dream than a reality. Advances in nuclear magnetic resonance (NMR) spectroscopy techniques, however, have now made possible the observation of mechanistic events at the molecular level of protein structure. Smith and colleagues, in an exciting contribution, review emerging ‘in-cell NMR’ techniques that demonstrate the potential to monitor biological activities by NMR in real time in native physiological environments. A current drawback of NMR as a structure determination tool derives from size limitations of the molecule under investigation and the structures of large proteins and their complexes are therefore typically intractable by NMR. A solution to this challenge is the use of selective isotope labeling of the target protein, which results in a marked reduction of the complexity of NMR spectra and allows dynamic processes even in very large proteins and even ribosomes to be investigated. Kerfah and co-workers introduce methyl-specific isotopic labeling as a molecular tool-box, and review its applications to the solution NMR analysis of large proteins. Tyagi and Lemke next examine single-molecule FRET and crosslinking following the co-translational incorporation of non-canonical amino acids (ncAAs); the goal here is to move beyond static snap-shots of proteins and their complexes and to observe them as dynamic entities. The encoding of ncAAs through codon-suppression technology allows biomolecules to be investigated with diverse structural biology methods. In their article, Tyagi and Lemke discuss these approaches and speculate on the design of improved host organisms for ‘integrative structural biology research’. Our volume concludes with two contributions that resolve particular bottlenecks in the protein structure determination pipeline. The contribution by Crepin and co-workers introduces the concept of polyproteins in contemporary structural biology. Polyproteins are widespread in nature. They represent long polypeptide chains in which individual smaller proteins with different biological function are covalently linked together. Highly specific proteases then tailor the polyprotein into its constituent proteins. Many viruses use polyproteins as a means of organizing their proteome. The concept of polyproteins has now been exploited successfully to produce hitherto inaccessible recombinant protein complexes. For instance, by means of a self-processing synthetic polyprotein, the influenza polymerase, a high-value drug target that had remained elusive for decades, has been produced, and its high-resolution structure determined. In the contribution by Desmyter and co-workers, a further, often imposing, bottleneck in high-resolution protein structure determination is addressed: The requirement to form stable three-dimensional crystal lattices that diffract incident X-ray radiation to high resolution. Nanobodies have proven to be uniquely useful as crystallization chaperones, to coax challenging targets into suitable crystal lattices. Desmyter and co-workers review the generation of nanobodies by immunization, and highlight the application of this powerful technology to the crystallography of important protein specimens including G protein-coupled receptors (GPCRs). Recombinant protein production has come a long way since Peter Lobban's hypothesis in the late 1960s, with recombinant proteins now a dominant force in structural biology. The contributions in this volume showcase an impressive array of inventive approaches that are being developed and implemented, ever increasing the scope of recombinant technology to facilitate the determination of elusive protein structures. Powerful new methods from synthetic biology are further accelerating progress. Structure determination is now reaching into the living cell with the ultimate goal of observing functional molecular architectures in action in their native physiological environment. We anticipate that even the most challenging protein assemblies will be tackled by recombinant technology in the near future.

High performance liquid level monitoring system based on polymer fiber Bragg gratings embedded in silicone rubber diaphragm

Liquid-level sensing technologies have attracted great prominence, because such measurements are essential to industrial applications, such as fuel storage, flood warning and in the biochemical industry. Traditional liquid level sensors are based on electromechanical techniques; however they suffer from intrinsic safety concerns in explosive environments. In recent years, given that optical fiber sensors have lots of well-established advantages such as high accuracy, costeffectiveness, compact size, and ease of multiplexing, several optical fiber liquid level sensors have been investigated which are based on different operating principles such as side-polishing the cladding and a portion of core, using a spiral side-emitting optical fiber or using silica fiber gratings. The present work proposes a novel and highly sensitive liquid level sensor making use of polymer optical fiber Bragg gratings (POFBGs). The key elements of the system are a set of POFBGs embedded in silicone rubber diaphragms. This is a new development building on the idea of determining liquid level by measuring the pressure at the bottom of a liquid container, however it has a number of critical advantages. The system features several FBG-based pressure sensors as described above placed at different depths. Any sensor above the surface of the liquid will read the same ambient pressure. Sensors below the surface of the liquid will read pressures that increase linearly with depth. The position of the liquid surface can therefore be approximately identified as lying between the first sensor to read an above-ambient pressure and the next higher sensor. This level of precision would not in general be sufficient for most liquid level monitoring applications; however a much more precise determination of liquid level can be made by linear regression to the pressure readings from the sub-surface sensors. There are numerous advantages to this multi-sensor approach. First, the use of linear regression using multiple sensors is inherently more accurate than using a single pressure reading to estimate depth. Second, common mode temperature induced wavelength shifts in the individual sensors are automatically compensated. Thirdly, temperature induced changes in the sensor pressure sensitivity are also compensated. Fourthly, the approach provides the possibility to detect and compensate for malfunctioning sensors. Finally, the system is immune to changes in the density of the monitored fluid and even to changes in the effective force of gravity, as might be obtained in an aerospace application. The performance of an individual sensor was characterized and displays a sensitivity (54 pm/cm), enhanced by more than a factor of 2 when compared to a sensor head configuration based on a silica FBG published in the literature, resulting from the much lower elastic modulus of POF. Furthermore, the temperature/humidity behavior and measurement resolution were also studied in detail. The proposed configuration also displays a highly linear response, high resolution and good repeatability. The results suggest the new configuration can be a useful tool in many different applications, such as aircraft fuel monitoring, and biochemical and environmental sensing, where accuracy and stability are fundamental. © (2015) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.