bluuebunny/crossref_metadata_2025 huggingface dataset:
Phoneme, Grapheme, and the ‘Importance’ of Distinctions.
Authors: Rischel Jørgen
Published: November 2023

The Scandinavian runic inscriptions dating from before 700 AD are nearly all written in a graphic system comprising 21 to 24 runes (originally and when presented as an alphabet or ‘futhark’ it comprised 24 runes (see Fig. 14.1), but some of these, notably the p and the k, fell out of use rather early).
Diphthongization in Faroese1,2
Authors: Rischel Jørgen
Published: November 2023

The diphthongs of Faroese are interesting from a general linguistic point of view, both because the inventory is typologically remarkable and because a comparison of the proto-language (OWS) and Modern Faroese (abbreviated MF) reveals some very puzzling developments of this system. The phonetic changes involved are in part very difficult to explain, at least if the case of each MF diphthong is considered in isolation. It is the aim of this paper to demonstrate that the various diphthongizations and other sound changes which can be observed are manifestations of a thorough reorganization of the whole vowel system that took place in the Faroese language of the Middle Ages (here termed ‘Old Faroese’: OF).
Devoicing or Strengthening of Long Obstruents in Greenlandic
Authors: Rischel Jørgen
Published: November 2023

It is a well-known characteristic of West Greenlandic Eskimo that this type of Eskimo (unlike North Alaskan Inupiaq, for example) has complementarity of voiced and voiceless fricatives, short fricatives being voiced and long ones voiceless (in the new Greenlandic orthography, which is used in this paper. g and r symbolize voiced velar and uvular continuants, and gg and rr their long voiceless counterparts, whereas the corresponding labial set is distinguished orthographically as v versus ff). In accordance with the principle of feature distribution just stated we find that voiced fricatives are automatically replaced by voiceless segments in environments where they are long by true gemination (or by assimilation), for example iga- ‘to cook’ – iggavik or igaffik ‘kitchen’, neripput ‘they ate’ – nerripput ‘they shared a big meal’.
How much grammar does it take to sail a boat?
Authors: Gil David
Published: November 2023

Human languages are of much greater complexity than the communicative systems of great apes, dolphins, bees, and other animals.1 Similarly, human culture, technology, and civilization are also immensely more complicated than anything observed in other species, such as the ways in which chimpanzees fashion tools to crack nuts or fish for termites. Clearly, these two facts are related: comparing humans to other species leads inexorably to the conclusion that linguistic complexity is correlated with complexity in other, non-linguistic domains. But what exactly is the nature of this correlation?
Linguistic complexity: a comprehensive definition and survey
Authors: Nichols Johanna
Published: November 2023

The long-received truism that all languages are ultimately about equal in complexity has taken some serious criticism in recent years. McWhorter (2001a) shows that one set of languages (creoles) are systematically less complex than most languages. Gil (1994, 2001; and cf. Chapter 2 above) shows that a non-creole variety of Indonesian is strikingly devoid of most kinds of morphosyntactic specificity and complexity, and Gil (2008) shows that isolating languages vary considerably in aspects of syntactic complexity. Ross (1996, 1997) and Dahl (2004) show that the sociolinguistics of contact and acquisition can greatly affect the complexity and amount of irregularity in a language. Hyslop (1993) shows that complexity in one lexical area (the number of distinctions in spatial demonstrative systems) varies inversely not with some other grammatical or lexical factor but with the size of the speech community; and Kaius Sinnemäki, in Chapter 9, below, demonstrates a subtler relationship between community size and a specific grammatical domain. Shosted (2006) and Sinnemäki (2008) select two components of grammar that might be expected to exhibit inverse levels of complexity if overall grammars were equally complex, and they show that there is little or no evidence of a negative correlation.
Ontological Anti-Realism∗*
Authors: CHALMERS David J
Published: November 2023

The basic question of ontology is ‘What exists?’ The basic question of metaontology is: are there objective answers to the basic question of ontology? Here ontological realists say yes, and ontological anti-realists say no.
Health literacy – achieving consumer ‘empowerment’ in health care decisions
Authors: Edwards Michelle , Hill, Sophie , Edwards Adrian
Published: November 2023

The term ‘health literacy’ was first used in a monograph describing the impact of health information on the educational system, the health system, and mass communication (Simonds, 1974). Since the United States’ National Adult Literacy Survey (Kirsch et al., 1993) identified 90 million adults as having inadequate literacy skills to meet the needs of the 21st century, there has been a concern about people’s abilities to function in health contexts. The Organization for Economic Cooperation and Development (OECD) (Darcovich et al., 2000) reports results from the International Adult Literacy survey showing that in 14 out of 20 industrialized countries more than 15 per cent of adults have only rudimentary literacy skills, making it difficult for them to cope with the demands of the information age. The implications of these results for health have instigated a refocus on health literacy and its meaning.
Decision analysis – utility for everyday use?
Authors: Thomson Richard
Published: November 2023

The challenge in health care choices, from system-wide decisions on new treatments through to individual decisions on treatment options, is to find ways of supporting good – or better – decisions. Decision-analytic (DA) techniques can address questions about the use of treatment for groups of patients, for example to underpin health policy decisions, or might be used to guide the management of individual patients (Ginsberg and Lev, 1997; Lilford and Royston, 1998). Their use to support individual treatment choices, though, is far less developed. This chapter will describe the potential value of decision analysis in shared decision-making, critique the method with reference to both practical and theoretical concerns, and consider the implications for practice and research.
Decision aids and shared decisionmaking in inflammatory bowel disease
Authors: disease Anne KennedyInflammatory bowel , decision-making consumer involvement in
Published: November 2023

Crohn’s disease and ulcerative colitis are inflammatory bowel diseases (IBD) that affect about 240,000 people in the United Kingdom (Carter et al., 2004). The cause is unknown and current medical treatment is to ameliorate rather than cure. Many patients take long-term drug treatment to control their condition, but surgery is sometimes an option when such treatment becomes ineffective. Symptoms include bloody diarrhoea, abdominal pain, and weight loss, and often follow a relapsing course with periods of remission. Management guidelines (Carter et al., 2004; Wexner et al., 1997) state that patients should be provided with information on treatment options and choice, with recognition of the patient’s perspective, experience, and expectations. In this chapter, I will look at how this can be achieved.
Decision support in the treatment of aneurysms of the abdominal aorta
Authors: Stiggelbout Anne M , Kievit J
Published: November 2023

An aortic aneurysm is local dilatation of the aorta–the main artery supplying blood from the heart to all the major organs of the body–usually representing an underlying weakness in the wall of the aorta at that location. While the stretched vessel may occasionally cause discomfort, a greater concern is the risk of rupture, which causes severe pain, massive internal bleeding, and, without prompt treatment, quickly results in death. Abdominal aortic aneurysms, hereafter referred to as AAAs, are the most common type of aortic aneurysm. They are mostly detected as an incidental finding by ultrasound in the course of physical examination or imaging, e.g. for urological or bowel symptoms, although some screening programs are being initiated both in the United States and the United Kingdom and are being discussed in other Western countries. A Cochrane systematic review has shown evidence of a significant reduction in mortality from AAA (but not in all-cause mortality) in men aged 65–79 years who undergo ultrasound screening (Cosford and Leng, 2007).
Decision aids and shared decisionmaking in mammography screening
Authors: Barratt Alexandra , Austoker Joan
Published: November 2023

Mammography screening is offered in many developed countries to women aged approximately 50–69 years to reduce mortality from breast cancer. The main benefits and harms of mammography screening are outlined in Box 45.1 (Barratt et al., 2005a) (for relative frequencies of these benefits and harms see Appendix 45.1). Information about screening generally emphasizes benefits and generally understates the harms (Jorgensen and Gotzsche, 2006), and there is a need for balanced, evidence-based patient decision aids on this topic, especially for younger women (Elmore and Choe, 2007; Qaseem et al., 2007).
Shared decision-making in cardiovascular risk management: Experiences and challenges
Authors: Weijden Trudy van der , Steenkiste, Ben van , Loon Marije S Koelewijn-van
Published: November 2023

Primary prevention of cardiovascular disease (CVD) is focused on patients without CVD but at an elevated risk of CVD. Public knowledge about symptoms and signs of myocardial infarction and stroke, or its risk factors, is limited, even among more highly educated and experienced patients (Bachmann, 2007). CVD is perceived as an undetectable or ‘sneaky’ disease in its earlier stages (Angus, 2005).
Decision aids for MMR vaccination
Authors: Trevena Lyndal , Leask Julie
Published: November 2023

Measles is a highly contagious disease which is contracted by almost all exposed non-immune children and those affected bear considerable risk of serious illness or death (Table 48.1). Prior to the development of a vaccine for measles in the 1960s, there were approximately 135 million cases and over 6 million measles-related deaths around the world each year (Wolfson et al., 2007). By 1987, the World Health Organization estimated that this had fallen to 1.9 million and a new combined vaccine for measles, mumps, and rubella became available (MMR). Global health.
Quantitative Approaches to Spatial Conservation Prioritization: Matching the Solution to the Need
Authors: Ferrier Simon , Wintle Brendan A
Published: November 2023

There is a growing interest in quantitative approaches to spatial conservation prioritization, from conservation scientists and practitioners alike. Subsequent chapters of this book explore, in considerable technical depth, the state-of-the-art of such approaches and describe software tools commonly used for their implementation. Here we step back from the details of specific techniques and tools to consider the general nature and role of quantitative prioritization from the perspective of conservation planning, and related disciplines of environmental planning and management.
The Zonation Framework and Software for Conservation Prioritization
Authors: Moilanen Atte , Kujala Heini , Leathwick John R
Published: November 2023

Zonation is a recently developed approach and software for spatial conservation prioritization (Moilanen et al. 2005; Moilanen and Kujala 2006, 2008; Moilanen 2007). Zonation effectively addresses a maximum utility type of problem (Chapter 3), taking into account distributions of features, connectivity responses, priorities for features, and variable emphasis placed on highest local quality (Sections 15.4–15.7). One main output of Zonation is a hierarchic ranking of conservation priority for all cells through the landscape, which can be visualized as a zoned map (Section 15.3). Zonation can, for example, inform the user of where to place or expand conservation (or restoration) areas, but it can also be used for the evaluation of existing or proposed conservation areas (Sections 15.8 and 15.9). Zonation is characterized by the ability to function on relatively large raster grid maps, facilitating a direct link from remote-sensing data and statistical distribution modelling. Using terminology of Chapter 1, Zonation primarily supports priority ranking and optimal plan generation, but it can also be used for scenario evaluation via load of solutions developed elsewhere.
The Plastic World
Authors: Gratzer Walter
Published: November 2023

Over the course of the nineteenth century some organic chemists produced unfriendly, insoluble, sticky substances by rather uncontrolled reactions, which we now know to have been polymers and the precursors of plastics. The first marketable material seems actually to have been derived from milk. The process is said to have been invented in about 1530 by a Bavarian Benedictine monk, by the name of Wolfgang Seidel, but he probably got the idea from earlier recipes. It was based on collecting the curd from sour milk, by filtration, and soaking it first in hot water and then in lye (an alkaline extract of wood ash). The result was a tough, sticky mass, which would have consisted largely of the protein casein. Seidel was searching for a substitute for cow horn, which was used, amongst other things, for making windows for lanterns. The horn had to be heated in steam and then flattened and shaped while hot.
The New Age: Giant Molecules for the Twenty-First Century
Authors: Gratzer Walter
Published: November 2023

A great leap forward in the chemistry and technology of polymers came midway through the twentieth century. It arose out of many failed attempts to prepare a polymer from a monomer that differed from ethylene (H2C==CH2) only by an additional methyl group: This molecule is propylene. The polypropylenes that resulted when propylene was treated in the same way as ethylene were of very low molecular weight, and mostly sticky gums.
Aristotle’ S Notion Of Priority In Nature And Substance
Authors: Peramatzis Michail M
Published: November 2023

I would like to thank David Charles and Lindsay Judson for their invaluable criticisms, objections, and suggestions. Thanks are also due to Brad Inwood and an anonymous referee of Oxford Studies in Ancient Philosophy. Any mistakes made in the present paper are, of course, solely due to me. K. Fine, ‘Ontological Dependence’ [‘Dependence’], Proceedings of the Aristotelian Society, 95 (1995), 269–90 at 271. Fine notes that Aristotle’s account of ontological priority too is couched in similar terms (270). This remark, however, is not intended as an interpretational proposal.
A Glance at the Terrain
Authors: Dixson Alan F
Published: November 2023

The purpose of these essays is to explore the evolutionary origins of human mating systems, the physical determinants of sexual attractiveness, mate choice, and patterns of copulatory behaviour. These are ambitious goals, given that sexual behaviour does not leave a fossil record.
The Road to Truth
Authors: Dixson Alan F
Published: November 2023

Much more is now known about human origins and evolution than was the case almost 40 years ago, when I began to study primate reproductive biology. In the final essence, it is the fossil evidence of hominid evolution, reviewed briefly in the first chapter of this book, which provides the backcloth against which the sexual behaviour and reproductive anatomy and physiology of the extant primates may be viewed in true evolutionary perspective. This has been the approach adopted throughout this book, and the goal of this final chapter is to present overall conclusions concerning the origins of human mating systems, patterns of copulatory behaviour, and mate choice.
Why is sleep so important?
Authors: Stores Gregory
Published: November 2023

There have been various theories about the function of sleep. For example, some have emphasized its role in brain development and in learning and memory. No one theory accounts for all of the complexities of sleep in different species and at different ages. What is clear is that sleep is essential for recovery and restoration. Sustained effort leads to tiredness, fatigue, and the need to sleep. If sleep is persistently disturbed, you will be affected psychologically and perhaps physically. Loss of sleep or poor-quality (non-restorative) sleep leads to the build-up of a ‘sleep debt’. This increases the pressure to sleep in order to pay off the debt when the next opportunity to sleep becomes available. Repeatedly disturbed sleep progressively builds up the size of the sleep debt. Fortunately, it takes far less time to pay off this debt than it takes to incur it. In other words, a relatively short period of sound sleep is sufficient to offset the immediate effects of sleep loss.
Sleep problems and their underlying causes (sleep disorders)
Authors: Stores Gregory
Published: November 2023

Fact: Nothing could be further from the truth. Careful enquiry will almost always reveal the reason why you are not sleeping well. This could be any of number of things. Once identified, the cause should be able to be treated effectively. Much of what has been said already shows that it is important to find out which of the various possible causes is responsible for someone not sleeping well. Such causes range from a minor illness or change of circumstances that will soon improve to serious conditions requiring intensive treatment. As described in later chapters, there are many types of sleep treatments. Which one is most appropriate for a particular person depends on the underlying cause of their sleep problem. Unfortunately, the underlying cause might not be considered, even when professional advice has been sought. No one would say that difficulty breathing is an illness in its own right for which there is only one type of treatment, but something similar to this might happen in the case of difficulty sleeping, for example. That being so, sleeping tablets should rarely be prescribed for insomnia (see Chapter 8) or stimulant drugs for sleepiness (see Chapter 9) without regard to the many possible explanations and which of them applies in the individual case. The many forms of parasomnia (see Chapter 10) also call for different types of advice and treatment.
Not sleeping well (insomnia)
Authors: Stores Gregory
Published: November 2023

Everyone has the occasional night of sleeping badly without the need for help or advice. However, surveys have repeatedly shown that about 30% of young and middle-aged adults are affected often enough by insomnia for it to be a significant problem. This figure is much higher (at least 50%) in elderly people and also in some other groups who are at special risk of disturbed sleep (see Chapter 11). People who show a lifelong ability to function perfectly well on relatively little sleep (‘short sleepers’) are not considered to suffer from insomnia. It is worth repeating that the problem of insomnia cannot be considered a condition or diagnosis in itself. It is a symptom and its cause needs to be discovered in each individual. There are many possible causes of insomnia, which, for the most part, are different, depending on how long the problem has lasted (see box below). There may be more than one cause, particularly in elderly people.
Disturbed behaviour and strange experiences at night (parasomnias)
Authors: Stores Gregory
Published: November 2023

Sleep is not always a relaxed, uneventful state. Often, various changes in behaviour or experiences take place during or in the period closely related to sleep. Sometimes these events are worrying to the person experiencing the parasomnia. On the other hand, he may not know what is happening because he is asleep at the time. So-called ‘arousal disorders’ (see later this chapter) are of this type. The changes in behaviour or experience in the parasomnias can be very subtle, but others are dramatic and alarming to the individual or to anyone who witnesses them. However, especially in childhood, even dramatic events of this type do not usually mean that they are caused by something serious. The box below lists various parasomnias in relation to the phase of sleep with which they are usually linked and, therefore, the time of night when they usually occur (see also Figure 3.1). It is essential that the episodes are correctly diagnosed and not confused with each other because the causes and treatments are very different. Distinguishing one type of parasomnia from another requires attention to detail. Sometimes, special sleep studies are needed (see Chapter 6).
An English Malady?
Authors: Scull Andrew
Published: November 2023

George Cheyne was a Scottish physician, one of a parade of ambitious and talented young men who had moved south to make their fortune around the time of the 1707 Act of Union between the two kingdoms of England and Scotland. A wit and initially something of a wastrel, Cheyne had sought to build his practice among the socially superior by hanging around the London coffee houses, flattering his would-be customers, polishing his credentials as a modern follower of Newtonian notions, and eating and drinking himself to a gargantuan size. More than 32 stone, or almost 450 pounds at his peak fighting weight, Cheyne grew so corpulent that he could barely move, and so naturally set himself up as a diet doctor. Affluence and luxury, after all, set loose the appetites, and excess exacts a predictable price in expanding waistlines and that quintessential eighteenth-century disease, gout.
The Revels Office
Authors: Chambers E K
Published: November 2023

ONE of the ‘ standing ‘ offices which, from the general oversight exercised over them by the Lord Chamberlain, may also be regarded as ‘ offices outward of the Chamber’ was the Revels Office. This, in its fullest establishment, consisted of a Master, a Clerk Comptroller, and a Clerk, whose services it shared with the analogous Office of Tents, a Yeoman, and a Groom. It was of Tudor origin. The first mention of a Master of Revels is in a Household order of 31 December 1494. But the post appears to have been at this period a purely temporary one, conferred upon some existing officer of the Household, who had been selected to supervise and defray the expenses of the revels for a particular feast.
The Mask
Authors: Chambers E K
Published: November 2023

THE mask is not primarily a drama; it is an episode in an indoor revel of dancing. Masked and otherwise disguised persons come, by convention unexpectedly, into the hall, as a compliment to the hosts or the principal guests. Often they bring them gifts always they dance before them, and then invite them to join the dance. They bring torch-bearers and musicians, who light and accompany the choric evolutions. Their intention lends itself to elaboration by spokes-men or presenters, and to such spectacular decoration as a pageant or scene affords thus it readily assumes a mimetic setting.
Treatment of other lung tumours
Authors: Falk Stephen , Williams Chris
Published: November 2023

Mesothelioma is caused by exposure to asbestos. There is a current epidemic of mesothelioma. Treatment of mesothelioma is largely to control the symptoms. Mesothelioma Mesothelioma is a tumour that develops in the lining of the lung known as the pleura. At the turn of the twentieth century this cancer was virtually unheard of. At the moment there is an epidemic of this tumour due to a clear-cut relationship to previous asbestos exposure.
Other methods of treatment
Authors: Falk Stephen , Williams Chris
Published: November 2023

There are a number of treatments aimed at destroying cancer locally. They are not routine practice and are used in very carefully selected cases, usually in specialist centres. Despite huge developments in our knowledge of cancer, treatment for lung cancer remains firmly rooted in surgery, radiotherapy, and chemotherapy. Newer, more cancer-targeted agents are making some inroads into the palliative treatments of non-small cell lung cancer, but new approaches continue to be needed. All of the following approaches have their pros and cons and none is widely available.
Getting the best treatment
Authors: Falk Stephen , Williams Chris
Published: November 2023

There is a national priority to improve cancer treatments and outcomes. Your care will be coordinated by a specialist multidisciplinary team. Your GP can refer urgently to the specialist team if cancer is suspected. National targets exist to ensure that you are promptly treated. National guidelines exist to ensure that patients throughout the country have access to best care.
Muscle symptoms and myotonic dystrophy
Authors: Harper Peter S
Published: November 2023

Muscle symptoms in myotonic dystrophy are very variable. The diagnosis is often delayed or missed because myotonic dystrophy is not being considered as a possibility. Muscle stiffness (myotonia) is important for the diagnosis, but much less important than weakness for patients.
Children with myotonic dystrophy
Authors: Harper Peter S
Published: November 2023

The first indication that a new born child has congenital myotonic dystrophy can be problems with breathing and feeding. This is due to weak breathing muscles. It is difficult for parents who know little or nothing of the disease, but try to find out as much information as you can; this will help when making decisions about the care and treatment of your child. Childhood problems from myotonic dystrophy are virtually unknown in the rarer ‘type 2’ form.
Receptor Preparations for Binding Studies
Authors: Hulme E C , Buckley N J
Published: November 2023

From the point of view of technique in binding studies, receptor preparations fall into two major categories: First preparations in which the cell surface membrane remains locally or globally intact. These include: whole tissues, in particular thin slices whole cells membrane preparations Secondly, preparations in which the cell surface membrane is disrupted by the me of a solubilizing agent.
Purification and Molecular Characterization of Muscarinic Acetylcholine Receptors
Authors: Haga T , Haga k. , Hulme E C
Published: November 2023

Here we present the procedures developed in our laboratory for the solubilization, purification, and characterization of mAChRs from porcine brain and heart. At least five different mAChR subtypes in mammalian tissues are known at present and their amino-acid sequences have been determined (I - 3,26). However, they have not been fully characterized as regards their pharmacological properties and localization, and methods of sufficient specificity to permit the purification of a given subtype from the mixture often present in the tissue of interest have not been reported. The procedures described here can be applied to purify any of the mAchR subtypes.
Photoaffinity Labelling and Purification of the β-adrenergic Receptor
Authors: Benovic Jeffrey L
Published: November 2023

Catecholamines regulate a wide variety of physiological responses via their interaction with specific cellular receptors (1). The catecholamine receptors have been divided into two major types, α and β each of which have been additionally divided into two subtypes. The β1 and β2-adrenergic receptors are coupled to stimulation of the enzyme adenylate cyclase which regulates the intracellular level of the second messenger, cyclic AMP. Although very similar in function, the β1 and β2 receptors are distinct in structure and pharmacological characteristics. The α1 and α2-adrenergic receptors are even more distinct, being coupled to different second messenger systems. The α1 -receptor is coupled to stimulation of phosphatidylinositol turnover, which regulates the level of the second messengers diacylglycerol and inositol trisphosphate, while the α2 -receptor is coupled to inhibition of adenylate cyclase.
Purification and Molecular Characterization of the -γ-aminobutyric acidA Receptor
Authors: Stephenson F Anne
Published: November 2023

γ-Aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the mammalian central nervous system. GABA mediates its effects via the specific interaction with GABA receptors of which two major subclasses have been defined on both a pharmacological and a functional basis. GABAA receptors are ligandgated chlorideion channels and are sensitive to the competitive GABAA receptor antagonist, bicuculline, whereas GABAB receptors are bicuculline-insensitive but baclofen, a GABAB receptor agonist, sensitive. GABAB-receptor activation results in the interaction with a guanine nucleotide binding protein and subsequent modulation of adenylate cyclase activity, of Ca2+ and/or K+ channel activity. GABAA receptors have a complex pharmacology, which has greatly facilitated the understanding of their structure, whereas to date there is little information available on GABAB receptors other than, by analogy with other receptors that act via second messenger systems, that they are proteins consisting of a single polypeptide chain.
The efficacy of high dose recombinant interleukin-2 based immunotherapy in man
Authors: Rubin J T , Rosenberg, S A , Lotze M T
Published: November 2023

The conventional treatment of cancer includes chemotherapy, radiation therapy or surgery used singly or in combination. Recently, a fourth modality, immunotherapy, has been shown to be effective in a limited number of haematalogical malignancies, and recent success against certain solid tumours suggests that immunotherapy will enjoy wider application. 3−6 The evolution of immunotherapy has been delayed by the lack of effective reagents in sufficient quantity. Seminal demonstrations of tumour immunogenicity in murine models7-11 were followed by successful therapy of tumour-bearing animals using in vivo and later in vitro sensitized lymphocytes administered locally or systemically .-19 Early attempts to apply these principles to the therapy of cancer in man began in 1966, when Nadler and Moore cross-transplanted histologically similar tumours in 13 pairs of patients. The subsequent transfer of lymphocytes between these patients was reported to be associated with tumour regression in 6 of 23 patients with metastatic melanoma. No responses were noted in a smaller number of patients treated with in vitro sensitized lymphocytes. In 1968, Frenster and Rogoway21,22 reported tumour regression in two of five patients infused with autologous lymphocytes activated in vitro with phyto-haemagglutinin (PHA). In 1970, this technique was also successfully applied to the local therapy of subcutaneous nodules by Cheema and Hersh. A pervasive problem in all of these studies was the inability to generate large numbers of activated lymphocytes.
Analysis Of The Cell Cycle In Saccharomyces Cerevisiae
Authors: Futcher Bruce
Published: November 2023

The basic mechanisms of cell-cycle control seem to be similar in all eukaryotes. The yeasts S. cerevisiae and Schizosaccharomyces pombe are two of the simplest and most powerful model systems for studying eukaryotic cell cycle control. Progress has been rapid, and the lessons learned have often been directly relevant to mammals and other eukaroyotes. Some methods useful for studying the yeast cell cycle are described here. However, these can be applied only in the context of the usual techniques of yeast molecular genetics and cell biology (I, 2). Similar methods for S. pumbe are described in Chapter 5, while methods for preparing synchronous cultures of both yeasts are described in Chapter 2.
Structure of RNA
Authors: Arani Gabriele V , Pardi Arthur
Published: November 2023

Our understanding of RNA’s central role in many cellular functions has increased dramatically since the discovery of RNA enzymatic activity (1). The field of structural biology has also made tremendous advances in the last decade and X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy have provided three-dimensional (3-D) structures of many proteins. However, there has been much less progress in structure determinations of RNA, with a major limitation having been the production of sufficient quantities of RNAs of defined sequence. While there have been few new X-ray structure determinations of RNAs because of the difficulties in obtaining suitably diffracting crystals, advances in the methods for large-scale synthesis (2) and in spectroscopic techniques have made it possible to apply multidimensional NMR to the solution-structure determinations of RNA. Tremendous progress in NMR techniques permits solution-structure determination of nucleic acids at a level of resolution comparable to crystallographic studies. This chapter will review recent methodological developments and structural studies of RNA secondary and tertiary structure domains and Chapter 2 will discuss phylogenetic, chemical, and enzymatic probing methods that have been used to determine the structure of larger RNAs.
Structure and function of hnRNP proteins
Authors: Kiledjian Megerditch , Burd Christopher G , Gorlach Matthias , Portman Douglass , Dreyfuss Gideon
Published: November 2023

The primary transcripts of eukaryotic RNA polymerase II, termed hnRNAs or pre mRNAs, undergo a complex and highly-regulated series of events in the nucleus as they mature into functional, cytoplasmic mRNAs (1, 2). Prominent among these events are the excision of intrans, generation of the 5’- and 3’-terminal structures and export of the processed mRNA into the cytoplasm. Upon the emergence of the nascent transcript from the transcription complex and throughout its nuclear life time, pre-mRNA exists as a complex with a group of nuclear pre-mRNA-binding proteins and it is in this form that the RNA undergoes nuclear RNA-processing reactions. The RNA-binding proteins which associate with nuclear pre-mRNAs but are not stable components of other nuclear structures (for example, snRNPs (small nuclear ribonucleoprotein particles)) are known as hnRNP proteins (3). The structure formed upon the association of these proteins with a pre-mRNA molecule is known as an hnRNP complex (reviewed in Dreyfuss et al. (4, 5)). hnRNP com plexes, therefore, occupy a central position in the pathway of gene expression and the study of their structure and function is of great interest.
Control of hutnan imtnunodeficiency virus gene expression by the RNA-binding proteins tat and rev
Authors: Karn Jonathan , Gait Michael J , Churcher Mark J , Mann Derek A , Mikaelian Ivan , Pritchard Clare
Published: November 2023

Replication of the human immunodeficiency virus (HIV) is controlled by two viral proteins, the trans-activator protein (tat) and the regulator of virion expression (rev). These regulatory proteins play complementary roles in the HIV life cycle: tat stimulates transcription from the viral long-terminal repeat (LTR), whereas rev is required for the efficient cytoplasmic expression of the mRNAs encoding the structural proteins of the virus. This system of dual regulation is illustrated in Figure 1. In permissive cells, transcription of the integrated proviral genome is initiated by host cell transcription factors (1-3). Initially, a low level of small, multiply spliced mRNAs encoding tat and rev are produced (4). tat synthesized during the primary round of transcription establishes a ‘positive feedback loop’ which boosts tat and rev mRNA production 200-1000-fold (5-7). Once a critical rev threshold is surpassed, HIV gene expression switches away from the production of the mRNAs for the regulatory proteins towards the production of the unspliced or partially spliced mRNAs encoding the virion proteins (8-10).
Expression, purification and interfacial kinetic analysis of phospholipase C-γ1
Authors: Jones Gwenith A , Horstman Debra A
Published: November 2023

Phospholipase C-γl (PLC-γl) is an important signalling molecule. Activation of PLC-γl by receptor tyrosine kinases allows growth factors, such as EGF, to modulate the level of the second messengers inositol 1,4,5-trisphosphate (Ins(l,4,5) P  3) and diacylglycerol (1-9). In response to EGF, PLC-γl rapidly becomes tyrosine phosphorylated and translocates to the membrane (1, 2, 7, 10-17). Very little information is available concerning the structure/function relationships of PLC-γl; an exception is the finding that tyrosine phosphorylation of PLC-γl at Y783 increases PLC-γl PtdIns(4,5) P  2 hydrolysing activity in response to growth factors (3).
Baculovirus-promoted expression, purification, and functional assay of G-protein regulated PLC-β isoenzymes
Authors: Paterson Andrew , Filtz Theresa M , Harden T Kendall
Published: November 2023

Knowledge of the catalytic and regulatory activities of inositol phospholipid specific phospholipases C (PLC) is critical to understanding at a molecular level the transmembrane signalling response to many hormones, neurotransmitters, and growth factors. These ubiquitous enzymes catalyse release of the second messengers inositol 1,4,5-trisphosphate (Ins(l,4,5) P) and sn-1,2-diacylglycerol (DAG). Three structural families of mammalian PLC isoenzymes (PLC-β, -γ, and -δ) have been identified, and the PLC-[3 class of isoenzymes accounts for the signal transduction response to the largest group of hormones and neurotransmitters.
Uses of site-specifically modified RNAs constructed by RNA ligation
Authors: Moore Melissa J , Query Charles C
Published: November 2023

Recent development of efficient methods for constructing site-specifically modified RN As, especially those longer than permitted by complete chemical synthesis, has opened up many new experimental avenues to researchers interested in protein:RNA interactions. These include the introduction of radioactive labels and photocross-linking reagents for detecting proteins or other RNAs that interact with a particular site of interest, and the introduction of functional group modifications for structure-function probing at single or multiple nucleotide positions.
Proteolytic Enzymes: Nomenclature And Classification
Authors: Barrett Alan J
Published: November 2023

A landmark in the development of any field of study is the appearance of a sound system of nomenclature and classification for the objects with which it deals. The introduction of the Linnaean system for naming and classifying organisms in the eighteenth century and the invention of a system of nomenclature for enzymes in the 1950s were such key events, and their value has been obvious. Both nomenclature and classification are vitally important for intonations handling, allowing people to communicate efficiently, knowing that they are talking about the same thing, and to store and retrieve information unambiguously. A good system will also serve to highlight important questions, and thus prompt new discoveries.
Mass Spectrometry Of Proteolysis-Derived Peptides For Protein Identification
Authors: Kuster Bernhard , Shevchenko Andrej , Mann Matthias
Published: November 2023

Proteolytic enzymes are used in biological mass spectrometry for countless purposes ranging from, e.g. the solubilization of membrane proteins to epitope mapping and protein conformational studies. In particular, proteases are indispensable tools for peptide mapping and protein primary structure elucidation. Until a few years ago, the latter application was usually performed either by N terminal sequencing via the Edman chemistry or by HPLC separation of peptide mixtures again followed by Edman sequencing. This type of analysis was quite time consuming and protein amounts in the order of 100 pmol were frequently needed for ‘ internal sequencing’ . However, only much lower amounts are frequently available from biochemical preparations involving multiple purification stages, thus more sensitive characterization techniques are needed. MS has gradually become a viable alternative to Edman sequencing and now generally offers superior performance. Sub-picomole amounts of peptides can routinely be detected and partially sequenced by MS. In addition, the molecular weight. as obtained by MS, is a highly characteristic physicochemical property of a peptide and can often be used for the identification of known proteins in sequence data bases without the need for generating any peptide sequence.
In vitro transcription and characterization of transcription factors
Authors: Cooney Austin J , Tsai, Sophia Y , Tsai Ming-Jer
Published: November 2023

Once a cis-element and transacting factor have been determined to be important for regulation of transcription of a gene, gel retardation, DNAase I footprinting, methylation interference analyses, and in vitro transcription assays are generally employed for characterization of a factor during purification. The majority of transcription factors are present in very low quantities within a cell, making purification long and arduous, and requiring large quantities of starting material. Thus, the initial characterization of the factor is undertaken with partially purified material. This permits elucidation of vital information about the factor, such as the molecular weight, subunit composition, and interactions, if any, with non-DNA binding factors, before complete purification of the factor (1). This information is mostly derived from analysis of data from a combination of in vitro transcription assays and chromatographic and electrophoretic techniques, which will be covered in this chapter.
Cloning transcription factors from a cDNA expression library
Authors: Cowell Ian G , Hurst Helen C
Published: November 2023

The initial chapters in this book describe methods of investigating the DNA binding activity of a transcription factor and how to use these properties to purify the protein. However, further studies concerning the functioning of the factor in transcription can be extremely limited unless a cDNA clone encoding the factor is isolated.
Preparation of chromatin templates for transcription studies
Authors: Boyes Joan
Published: November 2023

Transcription in the eukaryotic nucleus takes place within a highly packaged chromatin environment. Both genetic and biochemical approaches have been used to investigate how this process differs from transcription of protein-free DNA in vitro. This chapter describes various methods by which chromatin templates are generated for biochemical studies and discusses the merits of these different templates for particular studies. The preparation of histones and mononucleosome templates is described in full experimental detail. Moreover, the use of mononucleosome templates as a model to investigate nucleosome disruption over promoters is discussed.
Virus isolation
Authors: Cann A J , Irving W
Published: November 2023

There are two reasons why the difficult, expensive, and potentially hazardous procedure of virus isolation may be undertaken. The first is for the diagnostic identification of agents associated with disease. The other is to enable some subsequent experimental manipulation of the virus to be performed, for example, to examine mechanisms of replication or to determine the effectiveness of potential antiviral strategies in vitro. The protocols described in this chapter are derived from those used in a routine diagnostic virology laboratory, where the aim of virus isolation techniques is to cultivate and identify viruses from a variety of different types of potentially infected biological material (Table 1). Culture of viruses in large quantities for further experimental study relies on exactly the same principles, that is, provision of a suitable cell substrate, inoculation with material known to contain viable virus, and incubation under appropriate conditions.
Antiserum production and monoclonal antibodies
Authors: McKeating J , Shotton C , Valeri M
Published: November 2023

Since 1975, when Kohler and Milstein first reported the immortalization of antibody-secreting cells by somatic cell hybridization, hybridoma technology has proved to be an invaluable tool in the isolation and characterization of a variety of intra- and extracellular antigens (1). The principle is to propagate a clone of cells from a single antibody-secreting B lymphocyte, so that a homogeneous preparation of antibody can be obtained in large quantities. Hybrid cells are formed by the fusion of B lymphocytes (isolated from the lymph nodes or spleen of immunized animals) with a B-cell tumour line, most commonly, from the same species. The resultant hybridoma, once selected for specificity, will secrete the required antibody and can be grown indefinitely. Monoclonal antibodies (mAbs) produced against specific virus antigens have been used extensively to define the structure, function of virus components, and their interaction with proteins of virus and host-cell origin. Furthermore, such mAbs can form the basis of assays for the diagnosis of virus infection. Viruses generally have a regular, repeated, surface structure, and their surface-expressed antigens generally have a critical role in the attachment of virus to the host cell. Hence, such antigens provide a prime target for the immune system, and antibodies produced in response to these antigens have the potential to neutralize virus infectivity. These antibody-antigen interactions
Plant virus culture
Authors: Rybicki E P , Lennox S
Published: November 2023

Plant viruses are distinct from other major host-related groupings of viruses, such as animal or bacterial viruses, in that they are mostly single-stranded messenger-sense ss(+)RNA viruses, and mostly have simple, non-enveloped virions. There are a large number of types of simple isometric viruses, such as the Bromoviridae, Tombusviridae, and Comoviridae, as well as a wide variety of flexuous filamentous or rodlike types with helical capsid symmetry, such as the Potyviridae, Closteroviridae, and Tobamoviruses (1). There are some exceptions, notably with virus families that span different groups. Thus there are the enveloped ss(-)RNA plant Rhabdoviridae and Bunyaviridae, and the naked isometric dsRNA-containing plant Reoviridae (all of which may replicate in insect vectors and in insect cell cultures); there are two distinct groups of naked isometric ssDNA viruses with circular genomes (Geminiviridae and Nanoviruses) and two distinct groupings of plant pararetroviruses (Caulimo viridae and Badnaviruses) (2). While plant viruses as a group may not be as generally labile as their animal counterparts, given fewer enveloped viruses, it may be seen that there is considerable diversity, meaning that culture methods and methods for purification are similarly diverse.
Investigation of RNA virus genome structure
Authors: Easton A J , Marriott AC , Pringle C R
Published: November 2023

RNA Viruses: A Practical Approach is wide ranging in scope, from emerging technology such as reverse genetics and retrovirus vectors, to money saving tips - how to make your own silica particles for high efficiency RNA extraction and liposomes for cell transfection.
Packaging of segmented and non-segmented RNA virus genomes
Authors: Barr J , McCauley J W
Published: November 2023

The process of virus assembly describes the events by which the mature virion is formed from its constituent components within an infected cell. Because viruses come in such a wide variety of structural arrangements and consequently have adopted incredibly diverse assembly strategies, this definition is by necessity very broad. The range of processes included in this term ‘assembly’ are extremely varied and range from simple associations of a genome with no more than a few types of protein (Tobacco Mosaic Virus, TMV, or X174) to the formation of a virion comparable in size to a small bacterium and containing over 100 different proteins (poxviruses). A detailed description of assembly is beyond the scope of this chapter, or indeed this book, and so the reader wishing to know more about the various assembly strategies adopted