bluuebunny/crossref_metadata_2025 huggingface dataset:
chapter 7
Authors: Wu Duncan
Published: November 2023

Their engagement lasted longer than expected, John Stoddart’s prevarications dragging on into the new year of 1808. Lamb’s reaction was to invent the story that Hazlitt’s fretfulness had caused him to commit suicide by slitting his throat with a palette knife. That someone whose life was marred by tragedy could mount such a hoax seems astonishing until one considers that it was only by such gestures that Lamb maintained his own sanity. It prompted a robust response from the alleged suicidee, who penned a series of proofs demonstrating he was still extant, concluding he could see ‘no reason why [Lamb] should not be considered as much a dead man as himself, and the undertaker spoken to accordingly’.
chapter 10
Authors: Wu Duncan
Published: November 2023

Hazlitt had no difficulty finding work elsewhere. For Leigh Hunt he embarked on one of his finest theatrical reviews, of Kean’s Iago. Now at the height of his powers, Kean opened in the part on 2 July: it was, Hazlitt thought, ‘one of the most extraordinary exhibitions on the stage’. Instead of acting the villain, Kean performed him as ‘an excellent good fellow and lively bottle-companion’. This ‘completely foiled the critics’ who hardly knew what to make of it. But the audience did: they were spellbound.
chapter 21
Authors: Wu Duncan
Published: November 2023

Hazlitt put some coins in the driver’s hand and got in the chaise. His new wife was smiling, and he smiled back.‘Where to, sir?’, asked the coachman. Hazlitt looked quizzically at the man. Where to? The question took him by surprise, for he had given no thought as to where they might spend their honeymoon, only to finding the church at Coldstream in the Border country. Looking round, he saw two pointed hills in the distance.
The G20 After Eight Years: How Effective a Vehicle for Developing-country Influence?
Authors: Martinez-Diaz Leonardo
Published: November 2023

To many observers, the creation of the Group of 20 Finance Ministers and Central Bank Governors (G20) in 1999 marked a watershed in global governance. After decades in which the most important decisions in global financial and monetary management had been the exclusive province of a small club of the three, five, or at most seven richest nations, a ministerial-level network had emerged, bringing together the world’s most advanced economies and some of the largest developing countries. Convened at the initiative of the USA and other industrialized countries, the network promised to be a powerful, yet inclusive forum for global economic management. For the most sanguine commentators, the emergence of the G20 heralded a new age of more inclusive economic governance.
The Commission for Africa: A View Through the Prism of Networks
Authors: Wickstead Myles
Published: November 2023

In 2004, British Prime Minister Tony Blair brought together 17 people to form a Commission for Africa. Their task was to define the challenges facing Africa, and provide clear recommendations on how to support the changes needed to reduce poverty. The initiative had echoes of the Brandt Commission, an international panel brought together by World Bank President Robert McNamara in 1977 to break through the international political impasse in North– South negotiations for global development and to formulate basic proposals on which global agreement would be ‘both essential and possible’. The Commission for Africa had no formal authority to make decisions. Like the earlier Brandt Commission, individuals participated in their personal capacity and independently rather than as official representatives. It was thought that the Commission could influence thinking and policy by sharing ideas, furthering knowledge, and disseminating a new set of proposals through the wide networks of the members of the Commission. The Commission would be, in effect, a network of networks, whose members would learn from one another, generate a consensus among themselves, and use their official positions to press for action.
Oil and Gas in Africa
Authors:   ADB
Published: November 2023

The book, a joint work of the African Development Bank and the African Union, presents a comprehensive analysis of the oil and gas resources in Africa. It uniquely highlights, through country examples, and with an African focus but a global perspective, the specific challenges and constraints facing the continent as a whole in the exploitation and utilization of its oil and gas resources. It partly draws on a model that simulates the impact of high oil prices on African economies, a model that was developed by the Research Department of the Bank in a separate study. The roles of AfDB and AU are analyzed, considering their differing, but complementary, mandates geared towards the development of the continent. Finally, the book includes recommendations on the future directions and actions for maximizing benefits of Africa's oil and gas resources.
Maximizing the Benefits from Africa’s Oil and Gas Resources
Authors:   ADB
Published: November 2023

Africa is blessed with vast natural resource-rich environments. These resources constitute a principal source of public revenue and national wealth. Crude oil and natural gas, in particular, contribute significantly to the economies of resource-rich African countries (see Chapter 2). Under the right circumstances, an “oil bonanza” or natural resource boom can be an important catalyst for growth and development. Unfortunately, in many African countries natural resource booms have set off dynamic growth processes only to a limited extent.
Invasibility Analysis
Authors: Roff Derek A
Published: November 2023

An alternative approach to that used in the last chapter is invasibility analysis, which consists of asking if a clone displaying an alternate life history can invade a resident population. While one could compare results for markedly different life histories, in general, invasibility analysis has been used to locate the optimal combinations of parameter values rather than qualitatively different life histories. As with the “Fisherian” optimality approach, sexual reproduction is ignored. Invasibility analysis is used extensively, and is most useful, when fitness is density-dependent and there is age-or stage-structuring in the model. The method can handle stable, cyclical, and chaotic population dynamics. In this section I first consider the general structure of age- and stage-structured models and then describe the two general approaches of invasibility analysis, namely pairwise-invasibility and multiple-invasibility analysis. For all that you ever wanted to know about matrix population models see Caswell (2002).
Game Theoretic Models
Authors: Roff Derek A
Published: November 2023

Thus far, frequency-dependent interactions have been assumed not to be present, although they are undoubtedly important in nature (Clarke 1969, 1979; Endler 1986, 1988; Sherratt and Harvey 1993; Sinervo and Calsbeek 2006; Bond 2007). Putative examples of traits under frequency-dependent selection in natural populations are resource polymorphisms (Smith and Skulason 1996), mating polymorphisms (Sinervo and Lively 1996), color polymorphisms (Bond 2007), plant defenses (NÙñez-Farfa´n et al. 2007), and blood group antigen genes (Fumagalli et al. 2009). Analysis of models involving frequency-dependence is the domain of game theory. In a general sense, game theory is concerned with interactions between individuals: The basic scenario is one in which two individuals, called the players, meet and interact and either suffer a loss in fitness or an increase in fitness, called in either case the payoff. The important element of game theory is the Payoff matrix, which designates the increase or decrease in fitness to each player. Analysis consists in locating the Evolutionarily Stable Strategy (ESS), which, as previously noted, is defined as that strategy (or phenotype) which if adopted by all members of a population cannot be invaded by a mutant strategy (or phenotype) (Maynard Smith 1982). In general, game theoretic models are frequency-dependent but this is not an essential element of such models. Kokko (2007) gives an example of a frequency-independent model that involves the optimal growth rate of two neighboring trees. This case is considered in detail in Scenario 1. Here I shall examine the general elements of the analysis.
Radioimmunoassay
Authors: Edwards R
Published: November 2023

An immunoassay is undoubtedly the method of choice for the routine measurement of a peptide hormone. It has a potential for specificity and sensitivity. There are two basic types of immunoassay classified on the basis of relative reagent concentration (1). Immunoassay—antibody used at low or ‘limited’ concentration with a labelled antigen.
The molecular biology of polypeptide hormone expression and biosynthesis
Authors: Boam D S W , Clark A R , Shennan K I J , Docherty K
Published: November 2023

The techniques of molecular biology have made an important impact on our understanding of the regulation of polypeptide hormone expression and biosynthesis. Molecular cloning and sequencing of cDNAs encoding peptide hormones demonstrated that most are synthesized as larger precursors which undergo post-translational proteolysis to liberate the mature peptide. These precursors in many cases were shown to contain sequences for several peptide hormones or in some cases multiple copies of the same hormone. Different products could then be generated in different tissues. cDNAs have also served as probes for determining the site of hormone biosynthesis, and for studying the regulation of hormone gene expression.
The insulin receptor tyrosine kinase
Authors: White Morris F
Published: November 2023

Insulin controls metabolism and growth by modulating the activity of cellular enzymes and transport systems. The molecular events are initiated by insulin binding to the α-subunit of the insulin receptor, which stimulates tyrosyl autophosphorylation and activates a tyrosyl specific phosphotransferase in the β-subunit of the receptor (1). Thus, signal transduction may involve tyrosyl phosphorylation of cellular substrates by the activated insulin receptor kinase (2). Several putative substrates have been identified in the intact cell (3—6), including a protein which we call pp185 that is immunoprecipitated from insulin-sensitive cells with antiphosphotyrosine antibodies (7—9).
In situ hybridization and the three-dimensional reconstrt1ction of serial sections
Authors: Wilkinson David G , Green John
Published: November 2023

The advent of molecular biology has led to the identification of many genes with potential roles in the development of the mammalian embryo. These include homologues of Drosophila developmental genes; Multigene families of growth factors, receptors, kinases, and transcription factors; proto-oncogenes; and genes mapping to spontaneous or induced mutations. An important first step in the characterization of these genes is analysis of their pattern of developmental expression, particularly since this can yield clues as to their function. There are two major approaches used to determine sites of gene expression: the detection of protein by immunocytochemistry (see Chapter 7), and the detection of RNA by in situ hybridization. In situ hybridization takes advantage of the specific annealing that occurs between singlestranded nucleic acids of complementary sequence. The presence of a specific mRNA (designated by convention as sense-strand RNA) is revealed by using a labelled antisense probe of complementary sequence to hybridize a fixed tissue section. A major advantage of the in situ hybridization technique is the relative ease and speed with which specific probes can be produced from genomic or cDNA clones, or by the synthesis of oligonucleotides. However, determination of the sites of protein accumulation is also necessary in the longer term, in part because of the possibility that translational control occurs, and also since knowledge of the cellular location of the protein is important for further analysis of its developmental role.
Choice of an experimental animal an.cl roder1t genetics
Authors: Festing Michael f. W
Published: November 2023

The aim of this chapter is to offer practical advice on the choice of appropriate species of experimental animals for studies of postimplantation embryos, and on the choice of a suitable strain of animals once the species has been chosen.
Teratogen testing
Authors: Steele Christopher E , Copping Graham P
Published: November 2023

Chapters 1 to 10 of this book deal primarily with normal postimplantation development. In this chapter (see also Chapters 12 and 15) we consider the problem of abnormal embryogenesis. In humans, 65-70% of malformations are without known cause, approximately 20% result from genetic transmission, 3-5% from chromosomal aberrations, 4-6% from various extrinsic factors, and 3--4% from exposure to drugs and other chemicals (1). The substances responsible for the defects in this last category are known as teratogens and, although they are implicated in only a small proportion of the total malformations in humans, they are the most amenable to control. It is the identification and investigation of these substances that form the subject of this chapter.
Bacteria
Authors: Daniels Michael J
Published: November 2023

Some of the most serious diseases of crops are caused by bacteria. Although bacterial diseases can be found in all parts of the world, they are particularly important in tropical and subtropical countries where warm, humid conditions are ideal for bacterial growth. The pioneering work of Smith (1) nearly a century ago laid the foundations for our knowledge of plant pathogenic bacteria although these organisms have been somewhat neglected by plant pathologists until recently. With the emergence of ‘molecular plant pathology’, there has been an upsurge of interest in the major groups of plant pathogenic bacteria which represent excellent systems for detailed study.
Diffusion-limited growth
Authors: Wimpenny Julian W T
Published: November 2023

Molecular diffusion is a fundamental process of solute movement essential to all living organisms. Nevertheless, on a macroscopic level it is also a slow process. To produce fast predictable growth in laboratory fermentors, molecular diffusion is speeded up manyfold by incorporating eddy diffusion generated by agitators. In nature, the limitations of molecular diffusion are overcome for many systems in a similar fashion: in water bodies by convective, gravitational, and rotationally induced currents, by wave action and by the motion of living organisms themselves. In soils and sediments, percolation, the motion of gases and the activity of fauna and flora all assist molecular diffusion. Having said this, natural microbial ecosystems are quite unlike the simplified models used in most microbiological laboratories. Spatial heterogeneity and the general paucity of nutrients mean that molecular diffusion is the dominant transport process in a wide range of habitats. Examples of these include stratified water bodies, sediment ecosystems, the soil, and biofilm that forms on almost all surfaces exposed to appropriate levels of water and nutrients. These ecosystems have been reviewed in detail (Wimpenny, 1981; Wimpenny, 1989).
Protein Engineering of Antibody Combining Sites
Authors: Hilyard Kate L , Staunton David , Jones, Alison E , Rees Anthony R
Published: November 2023

1. Introduction The immunoglobulin G (IgG) molecule consists of a tetramer of two identical 25 kDa polypeptides (the light chain, L) and two identical 50 kDa polypeptides (the heavy chain, H). Crystal structures of IgG and IgG fragments have shown the antibody combining site (ACS) to be formed by the juxtaposition of six hypervariable loops or complementarity determining regions (CDRs), three from the light chain variable domain and three from the heavy chain variable domain. The CDRs of each chain are supported on a framework region which consists of conserved 13-strands that fold to form a β-sandwich. When a light and heavy chain come together, one surface of each sandwich associates to form a β-barrel structure (Figure la, b). Supported on this β-barrel scaffold the six CDRs, pack together in the tertiary structure to form a relatively flat platform with a surface area of about 700 Å2 (1). This variable domain, which contains all the determinants of antigen recognition, is one of six domains in the lgG molecule, the remaining five being Constant or C-type domains. The relative positions of these domains in the characteristic Y-shaped IgG and their nomenclature are shown in Figure lc.
Expression of Proteins in Yeast
Authors: Kikuchi Masakazu , Ikehara Morio
Published: November 2023

1. Introduction With the advent of recombinant DNA technology, it is now possible to express and produce many proteins in Escherichia coli (see Chapter 13). The coli expression system, however, has not always been found suitable for protein production, such as in the case of the hepatitis B virus surface antigen (1, 2). Further, post-translational processing and modification differ in E.coli and mammalian cells. Although mammalian cells can secrete biologically active proteins directly into the culture medium, genetic manipulation and cultivation on a large scale are not easy.
Emerging strategies for the determination of human carcinogens: detection, identification, exposure monitoring, and risk evaluation
Authors: Wright S
Published: November 2023

It is now generally accepted that most chemical carcinogens and probably all chemicals that induce cancer, de novo, i.e. cancer initiators, operate via a genotoxic mechanism. DNA is regarded as the key (primary and critical) target of genotoxic carcinogens although many chemicals classified as genotoxic are in fact inactive precursors requiring metabolic conversion into reactive forms (electrophiles) in order to undergo chemical reactions with DNA. Primary products of these reactions are generally promutagenic (or lethal) and their occurrence leads to an increased risk of mutation and cancer. Among the reaction products, DNA adducts have been singled out for special attention not only because of their mutagenic propensity but also because of the sensitivity and specificity of the methods for their analysis. DNA adducts and generalized DNA damage can also arise indirectly, e.g. via the agency of reactive oxygen species formed as a consequence of chemical or radiation-induced perturbations in normal cellular metabolism.
Haemoglobin adduct formation by acrylonitrile in rats and mice
Authors: Fennell T R , Macneela J P , Turner M J , Swenberg J A
Published: November 2023

Acrylonitrile (AN) is widely used in the manufacture of acrylic fibres and plastics. It is carcinogenic in the rat following long-term exposure either by inhalation or by oral administration, producing tumours in the stomach, brain, and Zymbal’s gland. No other species has been tested in a chronic bioassay. Epidemiological studies have provided limited evidence for acrylonitrile carcinogenicity in humans.
A comparison of P-postlabelling, immunoassay, and scintillation counting for the quantitation of benzo [β] pyrene adducts in mouse skin
Authors: King M M , Schell, C , Garner R C
Published: November 2023

Initiation of chemical carcinogenesis is thought to involve covalent binding of chemical carcinogens or their reactive metabolites to DNA, forming DNA adducts. Evidence suggests that carcinogenic potency of a number of polycyclic aromatic hydrocarbons (PAH) correlates with their ability to form specific adducts. Thus the detection of such adducts could be useful as markers of biologically effective dose.
Postlabelling of the DNA binding products of anticancer agents dacarbazine and cisplatin
Authors: Mustonen Riitta , Hietanen Paivi , Hemminki Kari
Published: November 2023

Several anticancer agents exert cytotoxicity by binding to DNA as their principle mechanism of action. However, limited information is available on the structural products of anticancer agents with DNA and measurements in humans have been carried out for cis-platin adducts only.
The detection of new genetic defects in man
Authors: Rossiter Belinda J F , Caskey C Thomas
Published: November 2023

A large number of genes involved in diseases are located on the X chromosome and many of the mutations at these loci are genetically lethal. It follows, therefore, that many of the mutations observed within these genes are new and constantly arising in the population.1 The continuous generation of new mutations provides a technical challenge for the efficient detection of a variety of genetic defects at these loci. Procedures using modern nucleic acid technology have been developed for the detection of such new and heterogeneous mutations, taking as examples Duchenne muscular dystrophy (DMD, dystrophin), Lesch-Nyhan syndrome [LN, hypoxanthine guanine phosphoribosyl transferase (HPRT) deficiency], and ornithine transcarbamoylase (OTC) deficiency. Each of these genes presents a different challenge for the detection of new mutations within it.
Contribution of occupation, diet, and smoking to PAH-DNA adduct load in peripheral blood cells
Authors: Strickland P T , Liou S H , Poirier M C , Nguyen D , Tockman M S
Published: November 2023

Polycyclic aromatic hydrocarbons (PAHs) are introduced into the environment of industrialized societies by a number of combustion processes. These include heat and power generation, coke production, open refuseburning and motor vehicle emissions. In addition to these general sources of PAH exposure, individuals can be exposed to PAHs through their occupation, diet, or smoking habits.
Lipoprotein separation and analysis for clinical studies
Authors: Mackness Michael I , Durrington Paul N
Published: November 2023

It is fitting to open this volume by considering clinical investigation since much of the impetus for research into lipoproteins in recent years stems from their likely relevance to atherosclerotic vascular disease. A prime purpose of the lipoprotein system is to transport lipids, but its theme has undergone a variety of evolutionary variations in different animal species. It is becoming apparent that lipoproteins have wider biological significance than simply lipid transport and are involved in such diverse processes as immune reactions (1), coagulation (2), and tissue repair (3). At the same time it is realized that lipoprotein metabolism may follow very different pathways in different species and that observations made in animals are frequently inapplicable to man. There is thus very often no substitute for clinical studies in man. Lipoproteins were first observed by the medieval physicians when venesection was popular and the milky serum associated with diabetes mellitus, nephrotic syndrome, and overindulgence in alcohol was well described. However, the discovery that this appearance was due to circulating fat particles secreted by the gut into lymphatics after fat absorption was not made until 1771 by Hewson (4). There followed the finding that fat was also present in fasting serum (5), the discovery of cholesterol (6), and early descriptions of the various clinical syndromes associated with hyperlipidaemia (7).
Analysis of tissue lipoproteins
Authors: Pitas Robert E , Mahley Robert W
Published: November 2023

This chapter describes in detail procedures we have used to analyse the lipoproteins present in cerebrospinal fluid (1) and tissue extracts (2) and to detect apolipoproteins in tissues (2). These procedures include extraction of apolipoproteins and lipoproteins from tissue and their subsequent analysis, as well as the detection of apolipoproteins within tissues and cells by immunocytochemistry. The coverage is not meant to be all-inclusive but rather to describe techniques that are in routine use in our laboratory and that may be applied to other systems.
PCR of TG microsatellites
Authors: Litt Michael
Published: November 2023

It is less than 10 years since the proposal was made to construct a genetic linkage map of man using restriction fragment length polymorphisms (RFLPs) (1). Since that time more than 3000 RFLPs have been described (2) and linkage maps have been published for many human chromosomes. Using these tools, numerous Mendelian diseases of man have been genetically mapped and it is generally agreed that, in principle, any human Mendelian trait may now be localized. However, certain practical considerations limit the application of linkage mapping in man.
The characteristics and growth of cultured cells
Authors: Butler Michael
Published: November 2023

In recent years, mammalian cells have become the preferred cell type for the large-scale production of a range of commercially valuable compounds (biologicals). The technology developed to exploit animal cell production is closely associated with traditional microbial fermentation processes used for bacteria and fungi. The characteristics of animal cell cultures are, however, significantly different from such microbial cultures. In general, growth is slower, the cells are more fragile, and the nutrient requirements are more complex.
Serum and its fractionation
Authors: Macleod Alex J
Published: November 2023

Media for maintenance of animal cells in vitro are usually aqueous solutions of a number of chemically defined low-molecular-weight components such as amino acids, vitamins, and salts, to which it is normally necessary to add a protein supplement. Animal blood serum is a well established medium supplement being a multifunctional reagent satisfying both nutritional and environmental requirements of the cells.
Growth factors
Authors: Jenkins Nigel
Published: November 2023

The proliferation of mammalian cells is not only governed by the availability of simple nutrients (as found in microbial cultures), but also by the actions of peptide growth factors found in serum. These factors bind to specific receptors on the cell surface which stimulate the cell into a new round of mitosis. The importance of such events (often mediated via protein kinases) is underlined by the fact that many oncogenes in cancer cells are related to growth factors or their receptors. Several growth factors also play a role in controlling cell differentiation or in the uptake of essential nutrients.
Downstream processing: protein recover y
Authors: Lyddiatt Andrew
Published: November 2023

Biochemical recovery (or downstream processing to adopt the popular sobriquet) encompasses those operations necessary to isolate and purify target product molecules from source feedstocks to specifications defined by applications. In respect of studies of cell physiology and metabolism, mammalian cell culture can be an end in itself, but the majority of technologies introduced in this volume have been developed with molecular productivity in mind. Current and near-future capabilities for native and recombinant protein production in animal cell systems have been reviewed elsewhere (1, 2). Such products will ultimately be representative of the broadest classification of protein molecules including monomeric and multimeric associations, hydrophilic and hydrophobic characters, and varying degrees of conjugation with carbohydrate, nucleic acid, lipid, and metal groups. Current and future progress in protein recovery from mammalian cell culture will thus largely depend upon universal advance in the design and implementation of biochemical downstream processing. There are few special circumstances surrounding mammalian cell culture which need elevate the molecular recovery of its products to a state of special technical status.
Theoretical aspects of practical centrifugation
Authors: Spragg S Peter , Steensgaard Jens
Published: November 2023

This chapter provides the reader with the theory and numerical bases upon which the practices of centrifugation are based. It particularly emphasizes the quantitative features of practical aspects of centrifugation such as sedimentation and determination of the efficiency of rotors. An in-depth knowledge of the various mathematical aspects of centrifugation is not an essential prerequisite for the average person. Hence the reader can omit this chapter if the protocols required are given in subsequent chapters of the book. However, if it is necessary to devise new centrifugation procedures, then an understanding of this chapter will provide an insight into the different parameters which should be modified to define the optimum conditions for centrifugal separations. For the sake of clarity the number of equations in the text has been kept to a minimum and, where derivations of equations are new or not readily available, these have been included in Appendix A of this chapter; derivations of equations that have been widely published (e.g. derivation of the Lamm equation) are not given here but readers are provided with appropriate references where these derivations can be found.
Introduction
Authors: Roberts Gordon C K
Published: November 2023

Nuclear magnetic resonance (NMR) spectroscopy is based on the fact that atomic nuclei oriented by a strong magnetic field (2-14 Tesla) absorb radiation at characteristic frequencies (typically a few hundred megahertz, in the radiofrequency part of the spectrum). The usefulness of NMR to the chemist and biologist results largely from the fact that nuclei of the same element in different environments give rise to distinct spectral lines. This makes it possible to observe signals from individual atoms even in complex biological macromolecules in solution. The parameters that can be measured from the resulting spectra can be interpreted in terms of molecular structure, conformation, and dynamics.
Resonance assignment strategies for Small Proteins
Authors: Redfield Christina
Published: November 2023

Prior to 1982, assignment of resonances in the complex NMR spectra of proteins was achieved, using ID NMR, and was based, for the most part, on the assumption that the structure of the protein in solution was the same as in the available X-ray structure. The introduction of 2D NMR techniques such as COSY and NOESY in the early 1980s dramatically increased the resolution of protein NMR spectra and as a result the amount of information which could be obtained from these spectra also increased. In 1982 With rich and co-workers introduced a systematic method for the full assignment of the 2D NMR spectra of proteins which relied only on information about the amino acid sequence of the protein; this method is the sequential assignment procedure (1,2). This approach is now used by most researchers who apply NMR spectroscopy to the study of proteins.
Extracellular single and multiple unit recording with microelectrodes
Authors: Millar J
Published: November 2023

Action potentials across the membranes of neurons generate electric current flow in the tissue around the neurons. These currents in turn generate voltages in the tissue, and these voltages can be detected as extracellular ‘spikes’. Extracellular spikes from mammalian neurons in the central nervous system have a duration of between 0.2 and 20 ms, depending on the type of neuron and the bandwidth of the spike recording system. Amplitudes vary from the noise level of the recording electrode (see below) which may be as little as 2 µV RMS (root mean square value) up to several millivolts: The great advantage of extracellular recording for the neurophysiologist is that you can record the activity of neurons without having to impale and thereby damage them. Impalement may be inadvisable because it damages the cell, it may be extremely difficult because the cell is small, or it may be unstable because the tissue is moving due to mechanical pulsations from blood pressure or respiratory fluctuations. For these reasons most neuronal recording in anaesthetized animals, as opposed to brain slices or cell cultures, is done with extracellular recording.
Intracellular recording methods for neurons
Authors: Silinsky E M
Published: November 2023

In the late 1920s, botanists interested in the behaviour of large plant cells pulled heated glass capillary tubing to give 1-2 µm tips, filled the resultant pipettes with electrolyte solutions, and impaled Valonia, Chara, and Halioclystis cells to study their transmembrane electrical properties (1). The use of such pipettes in animal cells soon followed when Cole and colleagues (2) recorded intracellularly from embryonic heart cells in tissue culture. The creation of the true microelectrode (generally defined as possessing a tip diameter < 1 µm) is frequently attributed to Gerard, Graham, Ling, and colleagues in the 1940s (3), who first described the conventional double pull technique for creating electrodes of 0.5 µm or less. The methods of Gerard and his associates also formed the link between the earlier artistry of those who pulled microelectrodes by hand and our present technological era of microprocessor-controlled electrode pullers. The diligence of these electrophysiological pioneers, coupled with post-war improvements in electronic circuitry, laid the groundwork for the next 30 years in which intracellular microelectrodes dominated the field of electrophysiology.
Computer-assisted recombinant DNA design and analysis with Matilda
Authors: Isralewitz Barry , ShallowAy* David
Published: November 2023

Creating a recombinant DNA usually involves two phases of analysis. Functional sites (e.g. boundaries of coding regions, intrans, promoters, etc.) and restriction sites are mapped in the first phase, usually both by experimentation and by DNA sequence analysis. The functional and restriction site maps that are produced in this phase are used to devise the recombination strategy in the second phase. The distinction between the two phases of analysis is apparent in the different ‘languages’ that are used. While sequences, restriction enzyme patterns, and consensus sequences are described using the four-letter nucleotide code, DNA fragments and recombinant clones are described using restriction enzyme names and locations, functional information, and descriptions of the boundaries and ancestries of DNA segments in a clone (which regions came from pBR322, for instance). Many software packages are available that provide the variety of DNA sequence analysis procedures applicable to the first phase (homology searching, translation, secondary structure prediction, etc.) (1). However, these programs do not, in general, directly address the problems of the second phase. These are usually described, and best solved, in terms of the higher-level language of restriction and functional site maps, not in terms of nucleotide sequence. This is particularly true for cloning operations with DNAs that have only been partially sequenced and where additional experimental data is important.
Expert systems and their implementation in chromatography
Authors: Mulholland M
Published: November 2023

Artificial intelligence (AI), and expert systems (ES) in particular, have been the subject of much hype and media-built expectation. This is mainly due to the human factor of this technology. ES apply human terminology to non human things and use their own peculiar language of fuzzy logic, demons, rules, and frames. All this surrounds them with an aura of mystery. It is interesting to speculate on why ES receive so much attention: there seem to be several reasons. Firstly, they promise to make information technology (IT) and computer software more human and less algorithmic. This allows computers to tackle a whole new arena of human problems. The ability of ES to imitate real experts also raises expectations that these problem solvers are easier to use and learn from than conventional software. Secondly, workers increasingly need to access expertise from outside their own subject domain. This is particularly true for the modern biochemist, who is expected to be a combination of statistician, analytical chemist, information technologist, mechanic, plumber ... the list could go on and on. Finally, it is a well-known business maxim that a company’s most valuable asset is its personnel. ES can allow this experience and skill to be preserved within a company. Taken all together, these reasons can leave no doubt as to why there is currently a huge market for this technology.
Autosomal dominant inheritance
Authors: Young Ian d.
Published: November 2023

Well over 2000 disorders and polymorphic systems showing definite or probable autosomal dominant inheritance have been documented.1 As the name implies, these conditions are determined by genes at autosomal loci and are manifest in the heterozygote. The provision of risks for disorders showing autosomal dominant inheritance is straightforward if there is a clear family history and the disorder in question is characterized by complete penetrance, consistent expression, and a reliable means of diagnosing heterozygotes. The risk to each child of an affected individual will be 50 per cent, whereas the child of an unaffected family member will be at negligible risk, or, to be more precise, twice the mutation rate (µ). Unfortunately, in practice risk calculation is rarely as simple as might at first seem. Factors which may significantly influence the final risk are considered in the following pages.
Methods for studying myristoyl CoA:protein N-myristoyltransferase
Authors: Rudnick David A , Duronio, Robert J , Gordon Jeffrey I
Published: November 2023

Protein N-myristoylation refers to the co-translational attachment of myristic acid (Cl4:0) to the NHrterminal glycine residues of proteins which participate in diverse biological processes including signal transduction, protein phosphorylation and dephosphorylation, oncogenesis, and viral assembly (reviewed in refs. 1-3). This covalent modification has been shown to affect profoundly the function of at least some N-myristoylproteins. The myristoyl moiety can play a role in protein targeting, but appears to represent more than a non-specific membrane anchoring signal as different N myristoylproteins are targeted to the plasma membrane (for example, p6osrc), endoplasmic reticulum (for example, cytochrome bS reductase), and even cytosol [for example, the catalytic (C) subunit of mammalian cAMP (3’5’ cyclic adenosine monophosphate) dependent protein kinase, PK-A).
Oligodeoxynucleotides Containing Modified Bases
Authors: Connolly Bernard A
Published: November 2023

Several hundred modified bases have been described in the literature and outlining their incorporation into oligonucleotides would clearly occupy several volumes. This chapter by necessity therefore only describes the preparation of a small number of bases in use in our laboratory. Each modified base has an exocyclic group replaced by either hydrogen or sulphur. We have mainly used oligodeoxynucleotides containing these altered bases to probe protein-nucleic acid interactions. However several of them also have potentially useful spectroscopic and chemical photore activity properties suggesting further useful applications.
Purification and characterization of lipoxygenase from germinating barley
Authors: Doderer Albert , Kokkelink Ingrid , Veen Simone van der , Valk Betty E , Douma Anneke C
Published: November 2023

Via the lipoxygenase pathway, free fatty acids (FFAs) in germinating barley can be converted into a variety of secondary metabolites, including compounds considered to play a role in staling and off-flavour formation in beer. In the present study the key enzyme of this route, lipoxygenase has been investigated. Two isoenzymes from germinating barley have been purified. On Western blots both enzymes crossreact with pea lipoxygenase antibodies. They have the same molecular weight and pH optimum but they differ in their isoelectric point and in the product formed upon incubation with linoleic acid. Amino-acid sequence data indicate some homology with soybean lipoxygenases. Further characterization of the enzymes as well as the cloning of the corresponding genes are in progress.
A mechanism for sulfite production in beer and how to increase sulfite levels by recombinant genetics
Authors: Korch Christopher , Mountain Harry A , Gyllang Hans , Winge Michael , Brehmer Patrik
Published: November 2023

Sulfite is important for the flavour stability of beer. In low alcohol beers its production is often inadequate. During fermentation, yeast produces sulfite and releases it into the wort. Earlier, we described several wort components that affect sulfite levels in beer. Herein we present a model explaining the accumulation of sulfite during fermentation. By recombinant DNA methodology, several methionine biosynthetic genes, cloned in 2-micron-based plasmids, were transformed into laboratory and brewing strains. The amount of sulfite synthesized during fermentation was increased by 2.5 to 20-fold, depending on the strain, its genetic makeup, and the genes carried by the plasmids.
Influence of industrial brewing and fermentation working conditions on beer SO2 level and flavour stability
Authors: Dufour J -P
Published: November 2023

Beer SO2 is generally considered as a positive factor against the staling of beer. However, more and more people have contradictory opinions on the antioxydant efficiency of naturally produced SO2 by yeast compared to the addition of SO2 to the beer before bottling.
The control of gene expression in brewing yeast with anti-sense RNA
Authors: Vakeria D , Box W G , Hinchliffe E
Published: November 2023

Brewing yeasts are difficult to manipulate using “classical” genetic techniques due to their high ploidy. Thus it is often necessary to effect changes at several genetic loci, which is both time consuming and problematic. Here we examine the potential of anti-sense RNA as a means of regulating gene expression in brewing yeast. We describe how an anti-sense RNA to the ILV-2 gene of Saccharomyces cerevisiae can be used to mediate changes in the resistance of brewing yeast to the herbicide sulfometuron- methyl. The effect upon vicinal diketone production is discussed in relation to the more widespread potential of the technique.
Genetical and biochemical characteristics of hybrid strains of brewing yeasts
Authors: Guinova-Stoyanova T A , Lahtchev K L
Published: November 2023

The construction and investigation of brewing yeast with glucoamylase activity /GAA/ is described. In the preliminary works Sta+Pof meiotic segregants of Saccharomyces cerevisiae were isolated and crossed with brewing strains. Different approaches were used for further improvement of the abilities of the hybrids obtained. Several strains with increased GAA were selected.
Immobilized yeast reactor application in continuous secondary fermentation in industrial scale operation
Authors: Pajunen Esko , Grönqvist Anders , Ranta Bo
Published: November 2023

The immobilized yeast reactor system is applied in industrial scale to shorten the lagering time in lager beer production. The present industrial scale unit has been running continuously during weekdays (also weekends) since april 1990. The flow rate is 40 hl/h, the average process time is 2 hours giving an estimated annual lagering capacity about 250,000 hl with space requirements of 50m area and 6m height. In the paper technical, economical and analytical aspects of the immobilized yeast reactor system will be dealt with.
Direct spontaneous conversion of acetolactate into non-diacetyl substance in fermenting wort
Authors: Inoue T , Murayama H , Kajino K , Kamiya T , Mitsui S , Mawatari M
Published: November 2023

It is widely known that acetolactate, a precursor of diacetyl, is quantitatively converted to diacetyl under brewing conditions, whether new or traditional brewing methods are used.
An image analysis sytem for measuring beer foam stability
Authors: Hegarty P K , Cope R , Crompton I E , Wallach E R
Published: November 2023

A computerised image analysis system is described for measuring beer foam stability. Beer is poured and foam collapse is recorded with a video camera. This information is analysed by computer to produce a quantitative measure of the time taken for foam collapse to occur. This technique can identify reproducible differences between the foam stability of different beers and dispense conditions. The procedure satisfies the criteria of an “ideal” method for measuring foam collapse: beer is not degassed, foam generation is not abnormal and, most importantly, the quantity of foam is assessed directly, in the same way as it is judged by the customer. The practical application of this system is discussed.
Beer off-flavours originating from anaerobic spore-forming bacteria in brewery adjuncts
Authors: Stenius V , Majamaa E , Haikara A , Henriksson E , Virtanen H
Published: November 2023

Brewery adjuncts are sometimes contaminated by anaerobic sporeforming Clostridium species responsible for butyric acid production. Butyric acid may be produced during adjunct manufacturing, but Clostridia may also grow in the brewery during storage. About 75-100% of the butyric acid present in the original adjunct passes through the brewing process to the final beer, causing a rancid off-flavour. When the concentration of butyric acid was analyzed in various raw materials as well as in samples throughout the process, 30-150 mg/l was detected in contaminated adjuncts, 7-22 mg/l in storage tanks and 3-15 mg/l in final beer.
N-Nitrosamines in brewing
Authors: Smith N A
Published: November 2023

Microbial nitrate reduction is the principal route of N-nitrosamine formation in brewing. However, little consideration has been given to how different groups of microorganisms possessing different mechanisms of nitrate reduction influence N-nitrosamine levels. Thermophilic denitrifying bacteria isolated from mashes can reduce nitrate producing a cocktail of N-oxides rather than nitrite alone, yielding N-nitrosamines at rates several orders of magnitude higher than contaminating organisms found in fermentation. This explains why N-nitrosamine levels cannot always be correlated with either bacterial numbers or initial nitrate concentration and has allowed the production of these compounds in brewing to be more effectively controlled.
Levels of limit dextrinase activity in malting barley
Authors: Longstaff M A , Bryce J H
Published: November 2023

Limit dextrinase could have a profound influence on wort dextrin composition and total alcohol production. During 9 days germination, samples of barley varieties of differing dormancy were kilned or freeze-dried. Limit dextrinase activity was measured using red-pullulan from MegaZyme. The initial activity was zero in all barleys, but increased at a rate which correlated with the germination rate. The activity was enhanced by cysteine or dithiothreitol in the extraction buffer, and reduced by kilning. Limit dextrinase activity during mashing could be manipulated by the length of malting, kilning and the extent of reducing conditions in the mash.
The relative importance of polyphenol oxidase, lipoxygenase and peroxidases during wort oxidation
Authors: Bamforth C W , Clarkson S P , Large P J
Published: November 2023

Of the enzymes suggested by the literature to promote oxidation during mashing, it is demonstrated that lipoxygenase and polyphenol oxidase are unlikely to be of significance. They are relatively unstable and are likely to be substrate-limited. Conversely, peroxidases do seem to be important. Ten peroxidase isozymes have been demonstrated, of which several have been partially purified. Their properties (stability, pH-activity profile, specificity and kinetics) have been studied and compared with the conditions of a mash. It is apparent that peroxidation is limited by the availability of substrate (hydrogen peroxide) and not by the level of enzyme present.
Studies on the synergistic interaction of konjac mannan and locust bean gum with kappa carrageenan
Authors: A.Williams P , M.Clegg S , J.Langdon M , O.Phillips K NishinarlAnd G
Published: November 2023

Studies have been carried out to investigate the nature of the synergistic interaction between konjac mannan and locust bean gum with kappa carrageenan. Differential scanning calorimetry and electron spin resonance spectroscopy both indicated that a specific interaction occurred and this was much more pronounced for konjac mannan than locust bean gum. The interaction occurred at temperatures just above the gelation temperature for kappa carrageenan alone and the interaction temperature depended on solution ionic strength. Rheological measurements showed that the interaction corresponded to the onset of gel formation.