bluuebunny/crossref_metadata_2025 huggingface dataset:
Health and Risk
Authors: Alaszewski Andy
Published: November 2023

In this chapter, I review current research on health and health care. In the opening section I consider the importance of risk in the development of health care, especially the ways in which the medical profession has used it to establish a dominant role, and the development of epidemiology as the main method of health risk analysis. In the second section I examine the current risk research agenda and the ways in which this is shaped by health policy agenda.
Life Course, Youth, and Old Age
Authors: Vickerstaff Sarah
Published: November 2023

In this chapter we consider how theorizations of the risk society have impacted upon the sociological understanding of the life course, we take current analyses and controversies about ‘youth’ and ‘old age’ as examples to illustrate contemporary debates. The discussion is divided into six parts. First, we provide a brief introduction to the idea of the life course and its traditional conceptualization and how risks and uncertainties are understood in the area of life course studies.
‘Genuine Accents’
Authors: Clark Heather
Published: November 2023

If the Belfast poets needed, in Mahon’s words, to ‘insist upon a different court of appeal from that which sits in the South’, they would have to insist upon different poetic fathers, ones who had wrestled with the peculiarly Northern ‘schizoid’ identity to which Longley frequently alluded.The work of Kavanagh, Hughes, Frost, Hill, Larkin, Yeats, and Lowell, though deeply influential, could not provide a common point of origin. But John Hewitt and Louis MacNeice could: for all their differences, both poets had confronted a sense of displacement that was familiar to Heaney, Mahon, Simmons, Longley, and Muldoon. Hewitt’s strategy was to turn his attention to the region, while MacNeice fashioned himself a restless cosmopolitan.
‘The Twilight of International Morality’? Hans J. Morgenthau and Carl Schmitt on the end of the Jus Publicum Europaeum
Authors: Brown Chris
Published: November 2023

The relationship between Carl Schmitt and Hans J. Morgenthau was fraught with all kinds of tensions.1 Both were, at least in the 1920s, conservative legal theorists who were critical of liberal accounts of politics and the law. Schmitt was approximately 20 years Morgenthau’s senior, and initially the latter appears to have regarded him as a potential mentor, sending him a copy of his doctoral thesis which overed a friendly critique of Schmitt’s very influential Concept of the Political (1st edn. 1927).
Introducing insect conservation
Authors: Samways Michael J , New Tim R , McGeoch Melodie A
Published: November 2023

Insects are the most speciose class of animals, with nearly a million described species, and probably several times this number still to be discovered, diagnosed, and named, especially in the tropics (Grimaldi and Engel 2005). Their functional roles are enormous. They play major roles in virtually every terrestrial and freshwater ecosystem. The conservation of insects has major functional significance in maintaining the diversity of ecological processes on which all life ultimately depends. These roles are often difficult to quantify in economic terms. Nevertheless, in the USA alone, wild insects are estimated to be worth $57 billion per year by providing services such as controlling pests, pollinating flowers, burying dung, and providing nutrition for other wildlife (Losey and Vaughan 2006).
Designing sampling protocols for insect conservation
Authors: Samways Michael J , New Tim R , McGeoch Melodie A
Published: November 2023

Insects, by virtue of their variety, small size, and large number, do not readily lend themselves to being counted or their diversity quantified. They most often first have to be conned, cajoled, or otherwise extracted from their crevices and other hiding places using a selection of the techniques that are described in the next chapter. However, once the target group or taxon (for example ground-dwelling or aquatic insects, such as ants or aquatic beetles) and most appropriate collection method(s) have been selected, several other questions arise. Ecologists (and entomologists in particular) are almost never able to obtain a complete measure of reality, and must rely on samples to ‘represent’ that reality.
Measuring environmental variables
Authors: Samways Michael J , New Tim R , McGeoch Melodie A
Published: November 2023

Insects respond to a range of conditions around them. Many insects have precise requirements for food (such as a particular host plant or dung type) and less readily quantified environmental requirements, such as suitable microclimate. Their incidence, abundance and distributions are governed strongly by such features. These conditions (broadly, environmental variables, EVs) may be determined by other organisms (biotic variables, such as interactions with plants and natural enemies), the physical world (abiotic variables, such as temperature or stream flow), or a combination of both (such as litter depth). Abiotic and biotic variables may combine to strongly influence the seasonal activity patterns of insects, and favourable conditions dictate the rate and timing of such development.
Biodiversity and assemblage studies
Authors: Samways Michael J , New Tim R , McGeoch Melodie A
Published: November 2023

Ecologists and entomologists describe patterns in insect species richness, abundance, and distribution, and seek ways to explain these. Biodiversity encompasses the variety of life, including genes, species, communities, and ecosystems (Gaston 1996). Insect biodiversity therefore includes examining the morphological (such as sexual dimorphism and differences between life history stages) and genetic variation within species, differences in characters, abundance and function between species, and patterns of species richness, composition, and structure within and between communities.
Reconsidering the Case for Black Reparations
Authors: Valls Andrew
Published: November 2023

The issue of black reparations has received a great deal of attention in recent years. On the face of it, this development would seem odd. After all, we are more than one generation removed from the close of the civil rights era which ended de jure segregation and outlawed most forms of racial discrimination. Yet it is precisely the failure of policies adopted during the civil rights era to bring about racial equality that lies behind the attention that the issue of black reparations is currently receiving. The idea of black reparations provides a framework for advancing the stalled progress toward racial justice.
Further Trouble for Unsettled Waters: Attention to Gender in the Debate on Black Reparations
Authors: Benson Carolyn
Published: November 2023

In a recent article on the history of the black reparations movement, Martha Biondi raises what would seem to be a straightforward question. Given the lack of discussion afforded to the particular exploitation suffered by black women under slavery and Jim Crow in the literature on reparations, she asks, will pending reparative policies address the realities of systemic sexual violence against women and the abuses of reproductive freedom during slavery and its legacy?1 While straightforward in and of itself, however, this question seems to be grounded in a larger and ultimately more complex set of questions concerning the appropriateness of introducing gender and gender oppression as central categories of analysis in the discourse on black reparations.
How Should We Respond to the Cultural Injustices of Colonialism?
Authors: Bhargava Rajeev
Published: November 2023

Colonialism has been a recurrent feature of human history. Everywhere in the past, lands have been conquered and new settlements have emerged brutally. Marauders who came to new lands to plunder wealth stayed on after killing the original inhabitants and stealing their goods. All brutal invasions were unjust even when they were not recognized to be so but in most such cases of past injustices, circumstances have changed so drastically as to supersede them. The possible historic injustice inflicted on the people of parts of north India by the Mughal conquest in the early sixteenth century has certainly been superseded first because of the common suffering inflicted on both Muslims and Hindus by British colonialism and second by the birth of political democracy in India. These past injustices of conquest and ancient colonialisms that have long ceased are not the concern of the chapter.1
All Cats are Grey by Night
Authors: Wright Patrick
Published: November 2023

In June 1990 Roy Kerridge wandered through Dalston for the benefit of readers of his ‘In the City’ column in The Independent. After relishing the vivacity of the ‘buxom Nigerian shoewives’ (whose ‘elaborate head-ties’ have a picturesque way of ‘nodding’ in the sweatshops) and declaring ‘fascinating Kingsland Road’ to be ‘the best place for shoes outside Lagos’, this beady-eyed social observer went on to describe his ‘happiest discovery’ in the area. Passing through ‘a secret entrance’ that runs below ‘a narrow iron railway bridge’, he found himself in Albion Square: a gracious and unexpected place that, with its ‘church, inn, trees and opulent terraces’, looked ‘like a transplanted corner of fashionable Canonbury or Highgate’.
The Bow Quarter: Six Hundred and Seventy Luxury Flats in an Old Victorian Hell-House
Authors: Wright Patrick
Published: November 2023

1988 was absurdly crowded with historical anniversaries. No sooner had the beacons been lit for the Spanish Armada than William and Mary landed at Torbay and started marching towards London for the Glorious Revolution. Somewhere on the road they passed a raggle-taggle encampment of hippies, poets and radicals gathered in sad commemoration of their youth.
The European Party System after Enlargement
Authors: Schmitt Hermann , Thomassen Jacques
Published: November 2023

In the introductory chapter we argued that despite the lack of a truly European process of political representation, elections for the European Parliament (EP) can still serve as an instrument of linkage between the will of the European people and the political decision making at the European level if a number of conditions are met. The first condition is that European party groups are competitive, i.e., distinct, and internally cohesive with regard to major policy dimensions. This condition will only be met if national political parties belonging to the same party group in the EP present compatible policy programmes to their national electorates.
Enlargement and the ‘Normal ‘ European Parliament
Authors: Voeten1 Erik
Published: November 2023

How does enlargement affect politics in the European Parliament (EP)? A large body of research indicates that the EP increasingly functions like a ‘normal ‘ parliament. This statement refers not only to the body ‘s gradually expanding legislative powers but also to the observation that its politics have not become dominated by national divisions but rather by supranational partisan politics along left–right ideological lines (Attina 1990; Thomassen and Schmitt 1997; Hix 2001; Kreppel 2002; Noury 2002; Raunio 2002; Kreppel and Hix 2003; Alter 2004; Hix, Noury, and Roland 2005, 2006a, 2006b). These findings are important because competition between supranational parties in a common low-dimensional ideological
Trust in Political Institutions
Authors: Wessels Bernhard
Published: November 2023

The last chapter dealt with output support from a utilitarian perspective. In terms of the Eastonion framework (see introductory chapter), this chapter goes a step further up in the hierarchy of objects of support and explores the levels of trust in institutions at the national and the European level. Since the major concern of this book is representation and accountability, analysis will be restricted to institutions that are functionally concerned with both aspects, that is, parliaments and governments. More precisely, we will analyse the development of trust in the respective national parliament and the European Parliament (EP)
Political Thought and International Relations
Authors: Bell Duncan
Published: November 2023

Political realism dominated the field of International Relations during the Cold War. Since then, however, its fortunes have been mixed: pushed onto the backfoot during 1990s, it has in recent years retuned to the centre of scholarly debate. Despite its prominence in International Relations, however, realism plays only a marginal role in contemporary international political theory. It is often associated with a form of crude realpolitik that ignores the ethical dimensions of political life. The contributors to this book explore alternative understandings of realism, seeing it as a diverse and complex mode of political and ethical theorising rather than simply a "value-neutral" social scientific theory or the unreflective defence of the national interest. A number of the chapters offer critical interpretations of key figures in the canon of twentieth century realism, including Hans Morgenthau, E. H. Carr, and Reinhold Niebuhr. Others seek to widen the lens through which realism is usually viewed, exploring the writings of Martin Heidegger, Hannah Arendt, and Leo Strauss. Finally, a number of the contributors engage with general issues in international political theory, including the meaning and value of pessimism, the relationship between power and ethics, the purpose of normative political theory, and what might constitute political "reality." Straddling International Relations and political theory, this book makes a significant contribution to both fields.
Demographic and Factual Overview
Authors: Saggar Shamit
Published: November 2023

Western Europe, informed estimates conclude, includes between 12 and 15 million Muslims. In the UK alone, the official census-based total is 1.6 million. This figure derives from a self-description methodology originating in 2001. Internal growth through childbirth and a young population age profile will have lifted the total substantially before the next scheduled headcount in 2011. Inward migration will also have contributed to rising numbers. At the time of publication, most reliable quasi-official estimates cluster around the 1.8—2 million mark.
Aid and Power: A Comparative Analysis of the Country Studies
Authors: Whitfleld Lindsay
Published: November 2023

In Aid and Power, Mosley, Harrigan, and Toye argued that there was no or little conditionality on World Bank loans in the 1980s to some countries, not because of any explicit ultimatum by those recipients, but because of an unspoken understanding that their governments would not welcome detailed programmes of policy reform being imposed on them as well as an awareness in the Bank of the importance of maintaining a lending programme in these countries (Mosley, Harrigan, and Toye 1995: 78).
The G20: A Practitioner’s Perspective
Authors: Rubio-Marquez Vanessa
Published: November 2023

This chapter focuses on one of the most visible networks of developing-country officials—the Group of 20 Finance Ministers and Central Bank Governors (G20). Drawing on my experience as both Director General of International Financial Affairs at Mexico’s Ministry of Finance and as Secretary General of the G20 in 2003—positions that allowed me to experience the internal workings of the G20 first-hand—I examine the origins and achievements of the network. I conclude that the G20 has evolved into one of the most important bodies for ensuring international financial stability and argue that there are powerful reasons for keeping the network alive. To preserve its relevance over time, however, the network needs to have a clear mandate vis-à-vis other institutions, as well as a carefully designed agenda that promotes the interests of all its members and one that lends itself to the provision of concrete deliverables.
Africa’s G4 Network
Authors: Bah Khadija
Published: November 2023

This chapter examines the functioning of African political networks, two in particular—the New Partnership for Africa’s Development (NEPAD) and the African Peer Review Mechanism (APRM). Within each, there are several formal, institutionalized networks. These include the NEPAD Heads of State and Government Implementing Committee (HSGIC), the APR Forum, the NEPAD Steering Committee, the APR Panel of Eminent Persons, and various national networks in member states. This chapter will demonstrate that behind these formal networks is an informal, non-institutionalized network of the Heads of State of four of Africa’s most influential states, a group I call ‘Africa’s G4’. The group is comprised of Presidents Thabo Mbeki of South Africa, Olusengu Obasanjo of Nigeria, Abdelaziz Bouteflika of Algeria, and Abdoulaye Wade of Senegal. This group was the force behind the evolution, adoption, and implementation of NEPAD and its APRM. This chapter explores the extent to which Africa’s G4, working through NEPAD and the APRM, has been able to change African politics and influence the industrialized North not only to recommit to the continent’s development but to do so on the basis of a new equal partnership based on mutual accountability.
Networking of Senior Budget Officials
Authors: Matheson Alex , Schoch Mickie , Kraan Dirk-Jan
Published: November 2023

This chapter examines a network which has strengthened national policymakers through exchanges of knowledge and peer support. The main network—the Senior Budget Officials’ Working Party (SBO) of the Organization for Economic Cooperation and Development (OECD)—is one of the oldest and arguably one of the most successful of the OECD’s many technical networks. This chapter explores how it has worked and also how two groups of developing and transition economies—one in Eastern Europe and the other in sub-Saharan Africa—are creating their own networks of senior budget officials following the OECD SBO model. These networks are, respectively, the Central Eastern and South Eastern Europe Senior Budget Officials Network (CESEE) and the Collaborative African Budget Reform Initiative (CABRI). This chapter is therefore about the promise and the challenges involved in emulating developed-country networks and applying the model to a developingor transition-economy context.
Introduction
Authors:   ADB
Published: November 2023

Africa has considerable oil and gas resources that can help accelerate growth on the continent if used strategically. Although new resources are discovered progressively, they are not equally distributed; indeed, 38 African countries are currently net oil importers. High and volatile oil prices are thus a challenge for all of Africa; they represent an opportunity to be pursued for exporting countries and an obstacle to be tackled for importing countries. The broad objective of this report is to shed light on the key issues related to the social, environmental, and economic impacts of high and volatile oil and gas prices. This includes a discussion of Africa’s oil and gas status, from a worldwide perspective, and the continent’s major challenges and opportunities in the energy sector.
Challenges and Opportunities
Authors:   ADB
Published: November 2023

This chapter examines the challenges and opportunities for turning oil and gas resources into economic wealth to alleviate poverty in Africa. The discussion focuses on providing a platform that will allow for pursuit of the best opportunities in the future to fully harness wealth from oil and gas resources. At the regional level, the energy demand-supply balance situation in Africa is positive: Given the region’s total energy resource endowment, substantially higher oil and gas consumption can adequately be met with regional supplies—with ample surplus still maintained for export. Despite this seemingly optimistic outlook, the country and sub-regional realities reveal multidimensional issues and challenges confronting expanded access to oil and gas in Africa (AU, 2007).
In situ hybridization for the localization and quantification of peptide mRNAs
Authors: Emson P C , Nunez, D J , Davenport A P
Published: November 2023

In situ hybridization (or hybridization histochemistry) is the application of nucleic acid hybridization techniques to the detection of DNA or RNA sequences in cytological and histochemical preparations. The method is inherently more sensitive than other nucleic acid hybridization techniques which depend on the preparation of DNA or RNA samples from homogenized tissue, in that the signal may be detected within an individual cell or group of cells. Thus, it is possible, for example, to detect which cells in a brain nucleus are expressing a particular gene and also to use a combination of radioactive and non-radioactive probes to identify several gene transcripts in a single tissue section. The sensitivity of the technique is such that a few copies of an mRNA can be detected within a cell and with the use of suitable densitometric methods, it is also possible to make data assessment quantitative. The technique of in situ hybridization for detection of nuclear DNA will not be considered here, but details can be found in a companion volume (Nucleic Acids Hybridization) (I). In this chapter we will concentrate on the procedures for radioactive and non-radioactive in situ hybridization used in our laboratory for the detection of peptide mRNAs and we will discuss our attempts at quantifying the signals. For further discussion of the method, its limitations and alternative protocols, the reader should consult reviews (1—5).
Localization of peptide hormones by light and electron microscopy
Authors: Pow David V , Clark Anne
Published: November 2023

Immunocytochemical techniques can be used to identify specific peptides in tissues, cells, and cell fractions. The sensitivity of immunocytochemical methods (ICC) are principally limited by the affinity of the antibody and the preservation of the peptide. Unfortunately, retention of antigenicity and preservation of ultrastructure are frequently incompatible. Ideally, all specimens would be fixed in a medium suitable for light (LM) and electron microscopy (EM) and a common detection system used. Specimens prepared for EM can be examined with LM either at the pre-embedding stage or with I µm thick resin sections. In practice, it is easier to process separate specimens for each study since, for LM, preservation of subcellular organelles can be sacrificed for maximal antigen preservation, whereas EM demands excellent retention of structural features often at the expense of antigenicity. Thus balances must be achieved for each peptide and each tissue.
Receptor characterization
Authors: Treadway Judith L , Pessin Jeffrey E
Published: November 2023

Characterizing the structural and functional properties of peptide hormone receptors is generally an important first step in determining the molecular mechanism of hormone action and transmembrane signalling in intact cells. In this chapter we give the procedures for isolating and purifying peptide hormone receptors from various tissues and cells, as well as characterizing the physicochemical properties of these receptors. We have focused primarily on methodology for insulin and IGF-I (insulin-like growth factor-I) receptors, with the intent that this information can be applied to other peptide hormone receptor systems as well.
Cyclic nucleotides
Authors: Lakey Trevor , Pyne Nigel , Murphy Greg
Published: November 2023

Peptide hormones bind to specific receptors on the exterior of the cell, and activate particular signal transduction systems, thus eliciting a physiological response. One type of signal which has been shown to mediate the effects of many hormones on many cells types involves the production or degradation of cyclic nucleotides [primarily cyclic AMP (cAMP) and cyclic GMP (cGMP)] inside the cell.
Inositol Phosphate Second Messengers
Authors: Kirk C J , Morris, A J , Shears S B
Published: November 2023

The role of inositol lipid hydrolysis in receptor-mediated cell signalling was first suggested by Michell in the mid-1970s (1). However, the current growth in interest in this subject probably stems from the more recent realization that receptor-mediated hydrolysis of phosphatidylinositol 4,5-bisphosphate [Ptdlns(4,5)P2] generates two intracellular second messengers. Following receptor activation, this lipid is hydrolysed by a ‘phosphoinositidase C’ to yield diacylglycerol, which activates protein kinase C, and inositol 1,4,5-trisphosphate [Ins(l,4,5)P3], which mobilizes Ca2+ from an intracellular pool (see references 2 and 3 for reviews).
Thin-layer chromatography
Authors: Henderson R James , Tocher Douglas R
Published: November 2023

Thin-layer chromatography (TLC) has long been used for the separation of lipid mixtures into their component lipid classes. Despite the fact that the more modern technique of high performance liquid chromatography (HPLC) is finding applications in lipid analysis, several features still make TLC particularly useful for this purpose. For example, although sophisticated and expensive apparatus can be purchased for TLC, good results on a qualitative basis can be obtained relatively simply using the minimum of equipment. Analyses are performed rapidly and many samples can be analysed simultaneously alongside standards. The separated classes can be visualized and the chromatograms kept as a permanent record, or, alternatively, samples can be recovered for further analysis. The present chapter considers the practical aspects of employing TLC for the analysis of lipids. It concentrates mainly on TLC in the ascending mode using glass plates coated with adsorbent because this is the most common form used in lipid analysis. Detailed descriptions of the historical and theoretical aspects of the technique can be found elsewhere (1). TLC with flame ionization detection (TLC-FID) is also considered since this technique is becoming increasingly popular for the quantitative analysis of lipid classes.
In situ hybridization in plants
Authors: Jackson David
Published: November 2023

In situ hybridization techniques provide an exquisitely sensitive method to study tissue-specific or organ-specific patterns of gene expression.

The techniques described in this chapter and based on refs 1--4, have been developed to study the expression of several genes involved in the biosynthesis of pigment (anthocyanin) in different floral organs (petal, style, stamen) of Antirrhinum majus. The methods involve hybridization of tritium-labelled single stranded RNA probes to mRNA in sections of fixed and wax embedded plant material. Hybridization is visualized using photographic emulsion, where radioactive emissions produce latent images in the emulsion which form silver grains when developed. The silver grains are visualized by dark-field microscopy. The techniques are applicable to a wide range of plant tissue and to different probes, including those involved in plant-pathogen interactions. For example, these techniques have been used at the John Innes Institute to study:

(a) storage protein gene expression in developing pea embryos (A. Hauxwell, personal communication);

(b) tobacco rattle virus RNA in and around infection sites on tobacco leaves (V.
Modifications to phosphoinositide signalling
Authors: Conn P Jeffrey , Wilson Karen M
Published: November 2023

Activation of a wide variety of neurotransmitter and hormone receptors results in hydrolysis of membrane phosphoinositides. It is now clear that phosphoinositide hydrolysis is the first step in a multifunctional system employed by neurotransmitter receptors for signal transduction. A number of studies over the last several years have led to a relatively detailed understanding of the nature of this response. These studies have been authoritatively reviewed by many experts in the field (1,2).
Exogenous kinases and phosphatases as probes of intracellular modulation
Authors: Hubbard Michael J , Klee Claude B
Published: November 2023

Protein phosphorylation is a major mechanism of intracellular regulation in neural tissues, modulating a variety of enzyme activities and processes such as those involved in neurosecretion, protein and catecholamine synthesis. Since the phosphorylation state of a phosphoprotein is dynamic, determined by the relative activities of kinases and phosphatases, factors affecting these enzymes must be addressed. Experimental manipulation at the level of kinases and phosphatases is a powerful tool for investigating intracellular regulatory mechanisms involving protein phosphorylation.
The expression of neurotransmitter receptors and ion channels in Xenopus oocytes
Authors: Leonard John P , Snutch Terry P
Published: November 2023

The microinjection of exogenous RNAs into Xenopus oocytes allows the examination of a variety of types of excitability proteins in a standard test environment. These include ion channels, neurotransmitter receptors, pumps, and transporters (1-5). In addition to expression studies, the oocyte system has proven useful for the isolation of cDNAs encoding these molecules [including a kainate receptor/channel (6), the serotonin-lC receptor (7, 8), and a delayed rectifier K channel (9)]. Furthermore, the oocyte system is a popular choice for studying the structural basis of receptor and channel function using mutagenized cDNAs (10-12). The ability of Xenopus oocytes to efficiently and accurately translate foreign RNA has been described in a previous volume in this series (36). This chapter will discuss the necessary requirements to successfully express ion channels and neurotransmitter receptors in Xenopus oocytes.
Molecular approaches to the structure and function of the GABAA receptors
Authors: Stephenson F Anne , Duggan Michael J
Published: November 2023

The GABAergic system is ubiquitous in the mammalian brain. It is estimated that GABA may function at up to 50% of the synapses and thus it is involved in the central regulation of multiple physiological processes. The GABAa receptors are the most abundant mediators of GABAergic neurotransmission. They are GABA-gated chloride ion channels and they are now known to be members of the ligand-gated ion channel super-family. In addition, the GABAa receptors are the site of action for several classes of centrally-acting drugs which include the benzodiazepines, the barbiturates, some steroids and non-competitive antagonists of GABAergic neurotransmission such as the cage convulsant compounts and picrotoxin (1). Indeed, it was the coexistence of the binding site for the benzodiazepines on the GABAa receptor oligomers which led to the isolation of this protein to apparent homogeneity by benzodiazepine affinity chromatography.
Growth and morphology of Fusarium graminearum and other fungi in batch and continuous culture
Authors: Trinci Anthony P J , Robson Geoffrey D , Wiebe Marilyn G , Cunliffe’ Bryan , Naylor T William
Published: November 2023

In 1964, Rank Hovis McDougall PLC (RHM) decided to develop a microbial fermentation to convert wheat starch (a by-product of the production of wheat protein) into high-quality protein foods for human consumption. Filamentous fungi were chosen for this process because of their nutritional and organoleptic properties, and because mycelial biomass, unlike unicellular biomass, can be formulated into a convincing facsimile of fibrous foods such as meat (Figure 1) and poultry. In 1969, Fusarium graminearum (Schwabe) was isolated from soil and this strain (A 3/5) has been used for all subsequent production of mycoprotein. The fermentation process developed for biomass production involved growing A 3/5 in continuous culture at a dilution rate of c. 0.19 h-1. Using a 1.3 m3 stirred-tank reactor (STR), RHM was able to produce c. 50 t of mycoprotein per year. In 1980, after the production of a two-million-word report describing the results of a ten-year programme of toxicology studies, RHM was given clearance by the UK Ministry of Agriculture, Fisheries and Food to market mycoprotein foods for human consumption.
Design of Protein Structures
Authors: Sander Chris , Scharf, Michael , Schneider Reinhard
Published: November 2023

The design of protein structures, whether by variation of natural proteins or from scratch, is an essential first step in the development of newly engineered proteins. Designs based on theoretical and computer methods enter the protein design cycle of repeated steps of experimental testing and improvement, much as in other engineering disciplines. Success in protein design depends on how well we make use of this cycle and on how well we understand the principles of protein folding, both in terms of the underlying molecular physics and in terms of knowledge extracted from the data-bases of macromolecular sequences and structures.
Assay of lipoprotein lipase and hepatic lipase
Authors: Bengtsson-Olivecrona Gunilla , Olivecrona Thomas
Published: November 2023

Lipoprotein lipase (LPL) and hepatic lipase (HL) are two key enzymes in the metabolism and interconversion of lipoproteins (1-3; Table 1). The reactions occur at endothelial surfaces of blood vessels, where the enzymes are attached by interaction with polyanions, probably heparan sulphate proteo glycans anchored in the endothelial cell membrane (4). LPL appears all over the vascular bed of most extrahepatic tissues, though in varying amounts, while HL is localized at the liver and some steroid hormone producing glands. LPL acts mainly on the triglycerides of chylomicra and VLDL. The released fatty acids are taken up by nearby cells or are circulated in blood as albumin bound free fatty acids. LPL is under metabolic and hormonal control. HL acts mainly on smaller lipoproteins (HDL, IDL, chylomicron remnants). Its function is not yet fully understood. Some animal species, e.g. guinea pigs and cows, have little or no HL. HL is regulated by steroid hormones, but does not seem to be metabolically regulated.
Membrane traffic pathways polarized epithelial cells
Authors: Ellis Juliet A , Jackman Mark R , Perez Jorge H , Mullock Barbara M , Luzio J Paul
Published: November 2023

Epithelial cells maintain a selective permeability barrier between extra cellular environments and regulate the composition of the fluids on either side of the cell layer. They achieve this by a diverse array of specialized activities including ion transport, secretion, and transcytosis (1), which are dependent on the polarized distribution of proteins and lipids between the apical and basolateral plasma membrane domains of the individual cells (2). Establishment and maintenance of cell surface polarity requires the formation of intercellular contacts and junctions, correct cell-substrate adhesion, highly ordered cytoskeletal networks, and specialized protein and lipid plasma membrane traffic pathways (3). This chapter confines itself to such membrane traffic pathways in polarized epithelia and, in particular, describes methods to examine the sorting of proteins to the correct plasma membrane domains. We provide protocols to study two different polarized cell types; a human, colon derived, adenocarcinoma cell line with enterocyte-like morphology called Caco-2 (4), which can be grown on filters in vitro as a polarized cell layer, and rat hepatocytes (5), which can be studied in situ in rat liver. We also discuss data from the MDCK (Madin-Darby canine kidney), renal-cortex cell line (6), which can also grow in culture as a polarized cell layer and from which much of our present knowledge of membrane traffic pathways in epithelial cells is derived.
Sorting between exocytic pathways in PC12 cells
Authors: Cramer Louise P , Cutler Daniel F
Published: November 2023

Exocytic protein traffic in neuronal and endocrine cells is complicated by the presence of more than one route to the cell surface. In addition to the constitutive secretory pathway, regulated secretory routes are also present in these cells. Regulated secretion is characterized by the intracellular storage of secretory material which is only released from the cell following the application of an external stimulus. This is in contrast to constitutive secretion in which proteins are not stored inside the cell but are constantly released. The organelles acting as storage compartments in neuronal and endocrine cells are both striking and characteristic of the cell type. In endocrine cells, dense core granules (DCGs) store secretory proteins, peptides, and small molecules such as classical neurotransmitters. In neurons, synaptic vesicles store neurotransmitters but not proteins. The coexistence of both regulated and constitutive secretory pathways in one cell was demonstrated by Kelly and co-workers in a DCG-containing cell line (AtT20) derived from the anterior pituitary (1, 2). This demonstration implied that regulated secretory cells must be able to sort regulated secretory proteins and constitutive secretory proteins so that only regulated secretory proteins are targeted to DCGs during granule formation. Later the coexistence of both pathways was also demonstrated in other regulated secretory cells.
In vitro reconstitution of vesicular transport from the endoplasmic reticulum to the cis Golgi in semi-intact cells
Authors: Schwaninger Ruth
Published: November 2023

Proteins destined for the plasma membrane, lysosomes, and endosomes, or the extracellular fluid are transported between organelles of the exocytic or secretory pathway of eukaryotic cells by vesicles that bud from one compartment and fuse with another. A high degree of specificity for this sequential budding, targeting, and fusion of transport vesicles must be maintained, or the cell would rapidly destroy its highly compartmental organization. Progression of newly synthesized proteins along this pathway can be followed by observing the maturation of N-linked oligosaccharide chains of glycoproteins during their passage through the different organelles. This is possible as individual oligosaccharide-processing enzymes reside in particular subcellular compartments.
Lipid modifications involved in protein targeting
Authors: Masterson Wayne J , Magee Anthony I
Published: November 2023

Several contributions in this volume describe the role of protein sequence motifs in acting as determinants of targeting specificity. In the last 12 years or so a group of post-translational modifications of proteins with various lipid molecules has been identified in eukaryotic cells (1). In many cases these modifications have been shown to be essential for the subcellular localization of the proteins carrying them. Many intracellular proteins are now known to carry fatty acid modifications which serve such a purpose. The 14 carbon saturated fatty acid myristate (C14:0) is amide-linked to the amino-terminus of a range of proteins, exemplified by the pp6osrc (proto) oncogene product. Site-directed mutagenesis has been used to demonstrate that this acylation is required both for membrane localization and transforming activity (2). The myristoylation enzyme has been isolated and is highly specific for acyl chain length (3). Inhibitors which interfere with this pathway provide an attractive approach to specific chemotherapy of cancers and viral infections including HIV (4).
Measurements of sedimentation coefficients
Authors: Steensgaard Jens , Humphries, Steven , Spragg S Peter
Published: November 2023

The sedimentation coefficient (s-value) of a particle is, by definition, its sedimentation velocity in a unit gravitational field (1). It is a commonly-used measure of particle size, especially when a molecular mass cannot be determined or when it is not physically meaningful as in the case of ribosomal subunits or other subcellular components. The sedimentation coefficient (given in Svedberg units, S) is a useful measure of particle size and it is an important parameter for the optimization of centrifugal procedures and the characterization of biological particles. This chapter describes how sedimentation coefficients can be calculated in practice.
Separations in zonal rotors
Authors: Graham John
Published: November 2023

This chapter is primarily concerned with the general design and operation of zonal rotors, but it also illustrates the advantages of zonal rotors by quoting some specific separations. The name ‘zonal’ rotor is unfortunate inasmuch as it is sometimes confused with ‘rate-zonal’ centrifugation. Rate-zonal centrifugation refers to the fractionation of a mixture of different sized particles as they move down (or up) through a gradient under the centrifugal force generated by the centrifuge; each population of particles moving independently of the others as a discrete band or zone at a rate roughly proportional to their size. Zonal rotors were originally designed to maximize the resolution of this process but they can be used for any type of centrifugation except, usually, simple differential pelleting. They also offer the capacity which makes the handling of large volumes of sample relatively easy.
In vivo voltammetric methods for monitoring monoamine release and metabolism
Authors: Stamford J A , Crespi, F , Marsden C A
Published: November 2023

Electrochemical detection (ECD), coupled with high performance liquid chromatography (HPLC), has become the method of choice for the assay of trace amounts of amine neurotransmitters and their metabolites in brain tissue samples since its introduction in the seventies (1-3). In vivo voltammetry essentially involves implanting a miniaturized version of the HPLC electro chemical detector into the brain.
Productivity tools biochemistry
Authors: Beynon Robert J , Easterby Johns.
Published: November 2023

Most commercial software for microcomputers is designed to sell many copies, and thus to be of value to a great many users. In general, this software provides tools that can be shaped or extended to meet user needs. This contrasts with specific software written to perform just a single task; these ‘vertical applications’ are often written by individuals who cannot devote the resources that are needed to generate a fully-supported, powerful, widely applicable tool. Such specific software also has its own advantages, being aimed more closely to a specific task, but its drawback is the relatively high cost of production. General-purpose programming environments, ranging from databases to graphics packages and spreadsheets, offer powerful access to the capabilities of modern microcomputers, and the development of a vertical application within such an environment is often preferable to the development of a task-specific program in a high-level language. Such environments are often overlooked, perhaps because their potential is not immediately apparent. in biochemistry. It is not possible to be comprehensive, and it would be wrong to assume that mention of specific packages implies a general recommendation, although such packages have been of use in our laboratories. At best, we can illustrate some of the potential of such packages, and indicate some of the considerations that are relevant when considering the acquisition of such a package. The scope of the chapter is also restricted to two of the most common types of machine, IBM PC and compatible systems running MSDOS, and the Apple Macintosh series. One of the difficulties attendant upon the introduction of proprietary software is the need to input and use data derived from other packages and instrument control systems, or to output the data in machine-readable form for use by other software. Some discussion of the principles of data exchange is therefore warranted, although there are so many packages on the market that they cannot be covered individually.
Introduction to Risk Calculation in Genetic Counselling
Authors: Young Ian d.
Published: November 2023

Genetic counselling is widely accepted as an integral part of the management of every patient with an inherited disorder. This book describes in detail how the risk can be estimated or calculated for most counselling situations, using nothing more complex than a knowledge of the simple laws of probability and a pocket calculator. All patterns of inheritance are considered. Particular attention is paid to the segregation of balanced chromosome rearrangements and risk calculation in single gene inheritance taking into account factors such as reduced penetrance, late age of onset, consanguinity, and the extended family. Also covered at length is the use of linked markers, an area of rapidly increasing importance in carrier detection and prenatal diagnosis. The text has been written in a straightforward "user-friendly" style with liberal use of worked examples and will be of benefit to everyone involved in the provision and delivery of genetic counselling services.
Identification of a glycosyl phosphatidylinositol anchor on membrane proteins
Authors: Hooper Nigel M
Published: November 2023

The explosion in the number of membrane proteins anchored by a glycosyl phosphatidylinositol (G-PI) moiety is due primarily to the ease with which the presence of such an anchoring structure can be identified. The identification of a G-PI anchor on a protein does not have to rely upon direct structural determination (see Chapter 8). A number of techniques exist whereby the presence of such a structure can be characterized even if the protein is present in low abundance or is not readily purified. The susceptibility of a protein to release from a membrane surface by bacterial phosphatidylinositol-specific phospholipase C (PI-PLC) was the original indicator for the presence of a G PI anchor (1). This technique is still the initial method of choice. Purified, recombinant PI-PLC is now available commercially, and simple sedimentation of membranes or whole cells can be used to separate the membrane-bound and released proteins. However, some G-PI anchors are resistant to hydrolysis by bacterial PI-PLC, and alternative methods are required to identify, and/or confirm, the presence of such a structure. These more recent techniques include using the mammalian G-PI-specific phospholipase D (G PI-PLD) recently isolated from plasma as an alternative to bacterial Pl-PLC to demonstrate the release of G-PI-anchored proteins from the membrane. Also, several antisera are now available which recognize the G-PI anchors of unrelated proteins following endogenous or exogenous phospholipase C digestion.
Phospholipases that degrade the glycosyl-phosphatidylinositol anchor of membrane proteins
Authors: Low Martin G
Published: November 2023

An unusual and useful property of the G-PI-anchored proteins is their ability to be released from membranes by bacterial phosphatidylinositol-specific phospholipase C (PI-PLC) which degrades the anchor. In fact, as described in Chapter 5, this property provided the first indications for the existence of G PI anchors (1, 2). Because the functions of the G-PI anchor remain obscure there has been growing interest in the possibility that eukaryotic cells might regulate expression of G-PI anchored proteins by activation of endogenous G-PI-specific phospholipases (2). Thus from a practical point of view interest in these phospholipases is of two quite distinct types.
Chemical and enzymic analysis of glycosyl-phosphatidylinositol anchors
Authors: Ferguson Michael A J
Published: November 2023

Glycosyl-phosphatidylinositol (G-PI) membrane anchors are ubiquitous among the eukaryotes and are utilized by a wide variety of cell-surface glycoproteins. The G-PI anchor can be thought of as an alternative to the hydrophobic transmembrane polypeptide domain for membrane protein anchorage. Discussions of the biosynthesis and the known and possible functions of G-PI anchors are to be found throughout this volume and have been extensively reviewed elsewhere (1-3). This chapter will concentrate on structure determination.
2’ -O-Methyloligoribonucleotides Synthesis And Applications
Authors: Sproat B S , Lamond A I
Published: November 2023

This enzyme specifically cleaves RNA in RNA-DNA heteroduplexes (8). In addition, 2'-O-methyloligoribonucleotides are chemically more stable than either oligodeoxyribonucleotides or oligoribonucleotides and moreover are totally resistant to degradation by either RNA- or DNA-specific nucleases (7).