Recombinant protein production in yeast:methods and protocols

In the last few years, significant advances have been made in understanding how a yeast cell responds to the stress of producing a recombinant protein, and how this information can be used to engineer improved host strains. The molecular biology of the expression vector, through the choice of promoter, tag and codon optimization of the target gene, is also a key determinant of a high-yielding protein production experiment. Recombinant Protein Production in Yeast: Methods and Protocols examines the process of preparation of expression vectors, transformation to generate high-yielding clones, optimization of experimental conditions to maximize yields, scale-up to bioreactor formats and disruption of yeast cells to enable the isolation of the recombinant protein prior to purification. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Fatigue cracking from stress concentration in mild steel

Pulsating; tension fatigue tests have been carried out on edge notched specimens of a mild steel. An electrical potential drop technique was used to determine the number of cycles taken to initiate cracks and the rate at which the cracks grew across the specimen. The results could be described by the range of stress intensity factor, which for crack initiation was modified to take account of the notch root radius. Analysis of elastic stress distributions at cracks and notches and models of plasticity at crack tips are used to discuss the results. Limited evidence in the literature indicates that the fracture mechanics approach may provide a general description of crack initiation and growth in notched specimens, and a simple graphical method of calculating fatigue lives is described. The results are used to illustrate the effects of specimen size and geometry on the fatigue life of notched specimens. The relevance of the work to the assessment of the significance of defects in welds is discussed.

Project financing 8th edition

The eighth edition is a fundamental and essential update to the seventh edition published in 2000. This new edition examines a comprehensive range of existing and newer topics that are relevant to project financing in 2012 and explores current trends in the project finance and leasing industries. Contributors are experienced academics and practitioners. Since the first edition was published, the financial markets have undergone tremendous upheavals and many new structures and instruments have been created to meet the financing needs of business. This edition considers the wider world of project finance, applicable to such diverse situations as venture capital and leveraged buyouts, and using new approaches such as Islamic finance techniques. The eighth edition is an essential and over-due update to the previous edition published in 2000. The eighth edition updates a comprehensive review of financial and related topics which are relevant to project financing in 2012 and explores current trends in financial modelling of a project, risk management and the private finance initiatives. This is a comprehensive and practical book full of advice and tips for successful project financing, including leasing, offering a clear, easy to understand guide to a complex area with examples. The topic coverage is well organized and complete moving from the fundamentals to the more complex issues. There is an extensive glossary to support readers. Finally the use of 12 practitioner case studies brings many of these complex issues to life. This is the new edition of the clear, easy-to-understand industry-standard text on project financing. With a good overview of a broad area and using principles of project financing to explain complex structures, this book includes lots of examples and case studies (including Eurotunnel, Dabhol, multiple Paiton deals and other recent deals along with subsequent developments) to show the concepts in use, examine outcomes and to ensure you understand important issues such as effective project structuring and financing, financial modelling for project valuation, and risk management. Substantially updated and expanded to provide the latest developments in all aspects of project financing. An important manual reference, this book is a must-have for every project financier's desk. The text unites the domain of project financing with a wealth of project management techniques, supported by diagrams and charts and other pictorial features, where appropriate. All these supporting features facilitate a better understanding of the accompanying text for the reader. In many chapters there are diagrams to clarify the specific transaction structure discussed in the accompanying text. These diagrams enable the reader to get a very clear idea of the transaction structure, which is particularly useful where it is complex or unusual. There are also a number of checklists to assist stakeholders in the project and resource management of complex project financings. The new financial modelling chapters allow exploration of some of the pitfalls project models encounter, challenging the accurate replication of the project cash flows for stakeholders to evaluate. In the later new risk management chapters, worked examples are included to illustrate the techniques in practice. The new public private partnership/private finance initiatives chapter introduces readers to this new approach to public projects. References are made to useful websites throughout the text. Cases are included at the end of the main text to encourage examination of real-life examples of project financing in practice and also highlight specific issues of current interest. The book will be helpful to project finance sponsors, lawyers, host governments, bankers and providers of capital

FASTtrack Pharmaceutical Compounding and Dispensing

FASTtrack: Pharmaceutical Compounding and Dispensing focuses on what you really need to know in order to pass exams. Concise, bulleted information, key points, tips and an all-important self-assessment section which includes MCQs, case studies, sample essay questions and worked examples. Based on the successful textbook, Pharmaceutical Compounding and Dispensing, this FASTtrack book has been designed to assist the student compounder in understanding the key dosage forms encountered within extemporaneous dispensing. For this new second edition all the references to modern texts (for example, the BNF) have been updated, as well as labelling to reflect changes since the publication of the first edition. Some worked examples have been changed owing to the availability of pharmaceutical ingredients. Free access to online videos demonstrating various dispensing procedures is included. Are your exams coming up? Are you drowning in textbooks and lecture notes and wondering where to begin? Take the FASTtrack route to successful study for your examinations. FASTtrack provides the ultimate lecture notes and is a must-have for all pharmacy students wanting to study and test themselves for forthcoming exams.

Analysis of protein carbonylation - pitfalls and promise in commonly used methods

Abstract Oxidation of proteins has received a lot of attention in the last decades due to the fact that they have been shown to accumulate and to be implicated in the progression and the patho-physiology of several diseases such as Alzheimer, coronary heart diseases, etc. This has also resulted in the fact that research scientist became more eager to be able to measure accurately the level of oxidized protein in biological materials, and to determine the precise site of the oxidative attack on the protein, in order to get insights into the molecular mechanisms involved in the progression of diseases. Several methods for measuring protein carbonylation have been implemented in different laboratories around the world. However, to date no methods prevail as the most accurate, reliable and robust. The present paper aims at giving an overview of the common methods used to determine protein carbonylation in biological material as well as to highlight the limitations and the potential. The ultimate goal is to give quick tips for a rapid decision making when a method has to be selected and taking into consideration the advantage and drawback of the methods.

Atmospheric plasma treatment of porous polymer constructs for tissue engineering applications

Porous 3D polymer scaffolds prepared by TIPS from PLGA (53:47) and PS are intrinsically hydrophobic which prohibits the wetting of such porous media by water. This limits the application of these materials for the fabrication of scaffolds as supports for cell adhesion/spreading. Here we demonstrate that the interior surfaces of polymer scaffolds can be effectively modified using atmospheric air plasma (AP). Polymer films (2D) were also modified as control. The surface properties of wet 2D and 3D scaffolds were characterised using zeta-potential and wettability measurements. These techniques were used as the primary screening methods to assess surface chemistry and the wettability of wet polymer constructs prior and after the surface treatment. The surfaces of the original polymers are rather hydrophobic as highlighted but contain acidic functional groups. Increased exposure to AP improved the water wetting of the treated surfaces because of the formation of a variety of oxygen and nitrogen containing functions. The morphology and pore structure was assessed using SEM and a liquid displacement test. The PLGA and PS foam samples have central regions which are open porous interconnected networks with maximum pore diameters of 49 μm for PLGA and 73 μm for PS foams. (Figure Presented) © 2007 Wiley-VCH Verlag GmbH & Co. KGaA.

FASTtrack:pharmaceutical compounding and dispensing

This is a revision guide for students giving bullet points of basic information on pharmaceutical compounding and dispensing followed by questions and answers."Fast Track" is a new series of indispensable revision guides created especially for undergraduate pharmacy students.The content of each title focuses on what pharmacy students really need to know in order to pass exams, providing concise, bulleted information, key points, tips and an all-important self-assessment section which includes MCQs, case studies, sample essay questions and worked examples."The Fast Track" series provides the ultimate lecture notes and is a must-have for all pharmacy undergraduate students wanting to revise and test themselves for forthcoming exams.Based on the successful textbook, "Pharmaceutical Compounding and Dispensing", this book has been designed to assist the student compounder in understanding the key dosage forms encountered within extemporaneous dispensing.A Fast Track website will also be live at time of publication and will include MCQs, sample online content and much more.

Changes at the BCLA

As we welcome 2014 we say goodbye to 2013 and I must start with an apology to authors who have submitted papers to CLAE and seen a delay in either the review process or the hard copy publication of their proofed article. The delays were caused by a major hike in the number of submissions to the journal in 2012 that increased further in 2013. In the 12 months leading to the end of October 2011 we had 94 new paper submissions, and for the same period to the end of 2012 the journal had 116 new papers. In 2012 we were awarded an impact factor for the first time and following that the next 12 month period to the end of October 2013 saw a massive increase in submissions with 171 new manuscripts being submitted. This is nearly twice as many papers as 2 years ago and 3 times as many as when I took over as Editor-in-Chief. In addition to this the UK academics will know that 2014 is a REF year (Research Excellence Framework) where universities are judged on their research and one of the major components of this measure remains to be published papers so there is a push to publishing before the REF deadline for counting. The rejection rate at CLAE has gone up too and currently is around 50% (more than double the rejection rate when I took over as Editor-in-Chief). At CLAE the number of pages that we publish each year has remained the same since 2007. When compiling issue 1 for 2014 I chose the papers to be included from the papers that were proofed and ready to go and there were around 200 proofed pages ready, which is enough to fill 3½ issues! At present Elsevier and the BCLA are preparing to increase the number the pages published per issue so that we can clear some of this backlog and remain up to date with the papers published in CLAE. I should add that on line publishing of papers is still available and there may have been review delays but there are no publishing online so authors can still get an epub on line final version of their paper with a DOI (digital object identifier) number enabling the paper to be cited. There are two awards that were made in 2013 that I would like to make special mention of. One was for my good friend Jan Bergmanson, who was awarded an honorary life fellowship of the College of Optometrists. Jan has served on the editorial board of CLAE for many years and in 2013 also celebrated 30 years of his annual ‘Texan Corneal and contact lens meeting’. The other award I wish to mention is Judith Morris, who was the BCLA Gold Medal Award winner in 2013. Judith has had many roles in her career and worked at Moorfields Eye Hospital, the Institute of Optometry and currently at City University. She has been the Europe Middle East and Africa President of IACLE (International Association of Contact Lens Educators) for many years and I think I am correct in saying that Judith is the only person who was President of both the BCLA (1983) and a few years later she was the President College of Optometrists (1989). Judith was also instrumental in introducing Vivien Freeman to the BCLA as they had been friends and Judith suggested that Vivien apply for an administrative job at the BCLA. Fast forward 29 years and in December 2013 Vivien stepped down as Secretary General of the BCLA. I would like to offer my own personal thanks to Vivien for her support of CLAE and of me over the years. The BCLA will not be the same and I wish you well in your future plans. But 2014 brings in a new position to the BCLA – Cheryl Donnelly has been given the new role of Chief Executive Officer. Cheryl was President of the BCLA in 2000 and has previously served on council. I look forward to working with Cheryl and envisage a bright future for the BCLA and CLAE. In this issue we have some great papers including some from authors who have not published with CLAE before. There is a nice paper on contact lens compliance in Nepal which brings home some familiar messages from an emerging market. A paper on how corneal curvature is affected by the use of hydrogel lenses is useful when advising patients how long they should leave their contact lenses out for to avoid seeing changes in refraction or curvature. This is useful information when refracting these patients or pre-laser surgery. There is a useful paper offering tips on fitting bitoric gas permeable lenses post corneal graft and a paper detailing surgery to implant piggyback multifocal intraocular lenses. One fact that I noted from the selection of papers in the current issue is where they were from. In this issue none of the corresponding authors are from the United Kingdom. There are two papers each from the United States, Spain and Iran, and one each from the Netherlands, Ireland, Republic of Korea, Australia and Hong Kong. This is an obvious reflection of the widening interest in CLAE and the BCLA and indicates the new research groups emerging in the field.

Do educators need educating?

Actual text: I was recently at the Spanish College of Optometry biennial conference and attended a meeting of contact lens lecturers from around Spain and Portugal. We discussed various ideas, mainly about how to share good practice and improve standards. What came to my mind was ‘is there a blueprint for training trainers?’ Well probably not but there are many things that we need to acknowledge such as the way students learn for example. Many educators themselves were taught by lecturers who would write on a blackboard or use acetate on an overhead projector, then came the 35 mm slide era followed by the Powerpoint era. More recently there is a move towards a much more integrated approach of various teaching methods. At my university our contact lens and anterior eye lectures generally follow a format where a narrated Powerpoint lecture is uploaded onto our internal virtual learning environment. This narrated version of the slides is designed to give the didactic element of the topic. The students listen to that before attending an interactive seminar on that topic. The seminar is also recorded so that students can listen to that afterwards. The seminar is designed to give additional information, such as case reports, or to clarify key points or for live demonstrations. It is a good way of doubling the contact time with the students without imposing further on an already packed formal timetable as the students can work in their own time. One problem that we noticed with this approach was that attendance can vary. If the students feel that they will gain something from the interactive seminar then they are more likely to attend – exam tips usually win them over! At the Spanish meeting the educators decided that they wanted to have regular meetings. The industry colleagues in attendance said that they were happy to help but could not necessarily give money, but they could offer meeting rooms, pay for lunch and evening meals. They even said that that they were happy to host meetings and invite other companies too (except to manufacturing plants). In the UK the British Committee of Contact Lens Educators (BUCCLE) meets for one day on three occasions in the year. The American Optometric Contact Lens Educators (AOCLE) meets annually at a three day event. Both these organisations get some help from industry. BUCCLE usually has one of its meetings at a university, one at a company training centre/manufacturing plant/national headquarters and one meeting the day before the BCLA annual conference. BUCCLE usually has its pre-BCLA meeting in conjunction with the International Association of Contact Lens Educators (IACLE). So when educators meet what would they discuss; well probably the focus should be on education rather than actual contact lens knowledge. For example sharing ideas on how to teach toric lens fitting would be better than discussing the actual topic of toric lenses itself. Most universities will have an education department with an expert who could share ideas on how to use the internet in teaching or how to structure lectures or assessments etc. In the past I have helped with similar training programmes in other countries and sharing good practice in pedagogy is always a popular topic. Anyone who is involved in education in the field of contact lenses should look at the IACLE web page and look out for the IACLE World Congress in 2015 in the days preceding the BCLA. Finally, IACLE, AOCLE and BUCCLE all exist as a result of generous educational grants from contact lens companies and anyone interested in finding out more about should refer to their respective web pages.

The science of writing an article – or is it art?

Getting your work published is an important aim for anyone undertaking research. This paper described how to prepare your work for peer review. Some recommendations are detailed i.e ensure yourself that your work is of good quality, decide is which journal you want to submit it to, follow the required style and be structured. Finally, a few tips and the reviewer's checklist are reminded.

Contract law concentrate

Contract Law Concentrate is a high quality revision guide which covers the main topics found on undergraduate courses. The clear, succinct coverage of key legal points within a specific topic area, including key cases, enables students to quickly grasp the fundamental principles of contract law. Written by Jill Poole, an experienced teacher and examiner and author of Textbook on Contract Law and Casebook on Contract Law, the book focuses on the needs of students to pass their exams. A number of pedagogical features help with the preparation for exams and suggest ways to improve marks. This guide has been rigorously reviewed and is endorsed by students and lecturers for level of coverage, accuracy, and exam advice. Packed with essential information, key cases, revision tips, exam Q&As, and more, Contract Law Concentrate is also supported by extensive online resources to take your learning further (www.oxford.com/lawrevision/): · Test your knowledge with the multiple choice questions and receive feedback on your answers. · Revise the facts and discussions of key cases using the interactive flashcards. · Learn the important terms and definitions using the interactive glossary. · Check that you have covered the main points of a topic using the key facts lists. · Achieve better marks following the advice on revision and exam technique by experienced examiner Nigel Foster.

G protein-coupled receptors:essential methods

G-Protein Coupled Receptors (GPCRs) are not only the largest protein family in the human genome but are also the single biggest target for therapeutic agents. Research into GPCRs is therefore growing at a fast pace and the range of techniques that can be applied to GPCRs is vast and continues to grow. This book provides an invaluable bench-side guide into the best and most up-to-date techniques for current and future research on GPCRs. With contributions from leading international authorities, this book equips readers with clear and detailed protocols for both well-known and up-and-coming techniques along with hints and tips for success. All the methods have been tried and tested by leading international research labs and are presented in easy-to-follow stages along with a useful overview of each technique. This book is an essential resource for all researchers in molecular biology, biochemistry, pharmacology and for graduate students.© 2010 John Wiley & Sons, Ltd.