bluuebunny/crossref_metadata_2025 huggingface dataset:
The Idealist Straitjacket Ibsen’s early aesthetics
Authors: Moi Toril
Published: November 2023

When Ibsen arrived in Christiania in 1850, he found a fairly undeveloped artistic and theatrical world focused on one great project: that of forging a national culture. Romantic nationalism ruled unchallenged. Obsessed with national identity, Norwegian art and literature were churning out ideal Norwegian heroes and heroines. Poems, plays, and genre paintings featured themes and characters inspired by the Icelandic sagas, Norwegian folktales and folk songs, and, not least, Norwegian peasant life. Because Norway never developed feudalism, Norwegian peasants and farmers (den norske bonden) were considered the very incarnations of national independence, bearers of the proud traditions of the Viking past, miraculously unsullied by foreign influences for more than 500 years. It is a measure of Ibsen’s isolation in Grimstad that he chose a Roman—that is to say, a European and cosmopolitan—subject, rather than a national one for his first play, Catiline (1850).
Becoming Modern Modernity and Theater in Emperor and Galilean
Authors: Moi Toril
Published: November 2023

Emperor and Galilean is pivotal in Ibsen’s work, literally as well as figuratively. It stands in the middle of his production: twelve plays come before, twelve plays come after. This enormous double play, a world-historical prose tragedy, also stands alone at the center of Ibsen’s œuvre. Four years (an unusually long gap for Ibsen) separate Emperor and Galilean from its neighbor on either side. In 1869 Ibsen finished The League of Youth, an entertaining comedy about politics and inauthenticity. In 1873 he published Emperor and Galilean, and in 1877, Pillars of Society. Together these three plays constitute Ibsen’s transition from being the author of Brand (1866) and Peer Gynt (1867) to becoming the author of A Doll’s House (1869). Of these three, Emperor and Galilean is by far the most important for an understanding of Ibsen’s modernism, for here Ibsen develops not just a full-scale analysis of modernity in Europe, but a new understanding of what theater is and what it can do. It also marks the moment in which Ibsen breaks with Norwegian nationalism and defines himself as a European: the immodest subtitle “A World-Historical Play” says it all. After Emperor and Galilean, Ibsen’s Norwegians are Europeans.
The Art of Transformation Art, Marriage, and Freedom in The Lady from the Sea
Authors: Moi Toril
Published: November 2023

The middle play of the so-called Munich trilogy, The Lady from the Sea (1888) follows Rosmersholm (1886) and precedes Hedda Gabler (1890). Whereas Rosmersholm and Hedda Gabler end with suicide, The Lady from the Sea has a happy ending. For this reason this remarkable play has often been treated as one of Ibsen’s least convincing and most light-weight works. Errol Durbach rightly comments that “the positive ending of The Lady from the Sea has opened it to ‘charges of artistic deficiency, of being somehow not echt Ibsen’—defying, as it does, the stereotypical image of Ibsen as the prophet of doom, while affrming the bourgeois carp-pond of marriage and family and distressing those modern Romantics for whom the dénouement betrays the vigorous Romantic spirit”.
Origins of the Olympics
Authors: Instone Stephen
Published: November 2023

This chapter is not called ‘The Origins of the Olympics’. That would have introduced an inappropriate suggestion of definiteness, since the very subject- matter of the formation of both the Olympic festival and athletics events within it does not allow certainty, straddling as it does the interface of history with myth. As Aristotle said at the beginning of the Nicomachean Ethics, ‘It is a mark of the trained mind never to expect more precision in the treatment of any subject than the nature of that subject permits’ (NE 1. 3). And the ancient Olympic Games are complex because they embraced not just physical endeavour but also several other aspects of Greek life, for example, religion and politics.
Pindar, Place, and Performance
Authors: Carey Christopher
Published: November 2023

The emphasis in several contributions to this volume on the local or regional aspect both of athletes and athletics and of Pindar’s activities as a panegyrist offers an opportunity to revisit the issue of performance in order to tease out some of the variables between locations, occasions, and contexts. It is as well to start by confessing ignorance and stressing—if warning were needed—the speculative nature of much that follows. Facts are few. There is much about the physicality of performance we would like to know but cannot recover. The music can only be recreated by speculation.
Thessalian Aristocracy and Society in the Age of Epinikian
Authors: Stamatopoulou Maria
Published: November 2023

He comments on his great fortune, having been a victor himself and living to see a victorious son. Using a geographical metaphor and the world of the Hyperboreans as an example of the limits of human accomplishment,15 Pindar paves the way for the mythical narrative, which recounts the deeds of Perseus (lines 27–48, especially 31–6). At first glance the choice of Perseus as the protagonist of a myth for a Thessalian victor might appear surprising, especially given the very frequent references to famous Thessalian heroes, namely Peleus, Achilles, or Jason, in other odes, especially those for Aeginetan victors.16 However, Perseus is ‘genealogically’ linked to Herakles, who has already been said by Pindar to be the ancestor of the Aleuads.
Costs
Authors: Laporte Audrey , Dolinschi Roman , Tompa Emile
Published: November 2023

This chapter suggests some good practice procedures in the assessment of the costs of an intervention. We intend it to be applicable to the entire range of types of economic evaluation studies—cost–benefit analysis (CBA), cost effectiveness analysis (CEA), cost–utility analysis (CUA), cost-minimizing analysis (CMA), and cost–consequence analysis (CCA)—but caution will have to be exercised in each context. Economic costs are not immutable entities that can be easily identified by the inspection of accounts. As has been explained earlier (see Chapter 2) the underlying idea of a cost is that it is a monetary sum representing the most valued alternative use of a resource (i.e., the opportunity cost). Costs will always be contingent on the perspective of a study and on the nature of the decision [classic references are Alchian (1959, 1968)]. For example, what appears as a cost in a study from the perspective of a workers’ compensation board might be a transfer from a societal perspective; what is not considered a cost in a study examining only the short term consequences of a possible decision may be a cost in a similar study taking a long-term view. Maintenance costs will be a significant cost component in a decision to introduce and maintain an item of equipment, but not in a decision to acquire it and sell it shortly thereafter. A decision to implement an occupational health and safety (OHS) intervention will invariably embody decisions about the timing (sooner versus later) and the speed (faster versus slower) of its introduction. Both typically affect costs: generally, the sooner the implementation the greater the cost; the faster the implementation the greater the cost. What is always true, however, is that the costs to be considered are the resource used as a result of the decision in question.
Parasitism, biodiversity, and conservation biology
Authors: Lebarbenchon Camille , Poulin Robert , Thomas Frédéric
Published: November 2023

Biological diversity—or biodiversity—can be defined as the variety and variability of all living organisms. It includes the genetic variability within species and their populations, the variability of species and of their life-forms, the diversity of related species complexes and of their interactions, and that of the ecological processes that are influenced by the organisms or of which organisms are the actors. Besides systematics and phylogeny that describe and classify the diversity of living organisms, population and community ecology are two of the main subjects for ecologists who study the dynamics of biodiversity.
Born’s Probability Interpretation of Quantum Mechanics (1926)
Authors: Brandt Siegmund
Published: November 2023

Schrödinger’s wave mechanics was using the mathematics familiar to all physicists and was therefore preferred by most over matrix mechanics. Moreover, it contained a function Ψ, the Schrödinger function or wave function, which was expected to be connected in some way to the position of a particle in space. For some time this connection was unclear. Bloch quotes a verse (original and translation) by Hückel , recited in the summer of 1926 in Zurich, which describes the situation in a humorous way [1]:
Do We Know How Happy We Are?
Authors: Haybron Daniel M
Published: November 2023

Never mind what it’s like to be one of Professor Nagel’s bats. I want to know what it’s like to be me. This is a real question, not a joke, because there are good reasons for doubting that any of us have a firm grasp on the quality of our experience of life, in particular its affective character. Possibly, many of us are profoundly ignorant about such matters, to the point that we often don’t know whether we are happy or unhappy, or even whether is our experience is pleasant or unpleasant. This, at any rate, is what I will argue in what follows.
Technical and organizational prerequisites for ultrasound-guided nerve blocks
Authors: Marhofer Peter
Published: November 2023

Some important issues should be considered prior to the introduction of ultrasound-guided regional anaesthesia in daily clinical practice. It is of particular importance to highlight the necessity of considering technical and organizational requirements for the adequate and optimal use of this technique and its initial introduction.
Ultrasonographic appearance of nerves and other anatomical structures
Authors: Marhofer Peter
Published: November 2023

Adequate interpretation of all anatomical structures in ultrasonography is a prerequisite for the correct performance of regional techniques.

Recall that ‘every nerve has a particular visualiztion in ultrasonography’. This statement may sound confusing, but it reflects our observations during the past decade.
Lower extremity blocks
Authors: Marhofer Peter
Published: November 2023

The lumbar plexus derives from the ventral rami of the spinal nerves L1–L3 and parts of L4 (furcal nerve). The first lumbar ramus commonly receives a branch from Th12. The lumbar plexus is formed within the posterior portion of the psoas major muscle. The branches of the lumbar plexus are the iliohypogastric, ilioinguinal, genitofemoral, femoral, lateral femoral cutaneous, and obturator nerves.
Catheter techniques
Authors: Marhofer Peter
Published: November 2023

All catheter techniques can be performed as either single or continuous techniques: the degree of visualization of the nerves is equal. During positioning, the catheter could be directly visualized in most cases (Figure 12.1). More importantly, the spread of local anaesthetic was able to be observed. We suggest the use of adhesive, transparent tape for fixation of a peripheral nerve block catheter, so that a daily ultrasonographic investigation of the spread of local anaesthetic is possible.
A key to intersubjectivity: conjoint constitution
Authors: Berthoz Alain , Petit Jean-Luc , Macann Christopher
Published: November 2023

This chapter deals with the question of our relations with others: intersubjectivity. For a century or so, physiology has mostly been a physiology of the perceiving and acting subject. Philosophers might call it a solipsistic physiology. During the same period, social psychology has kept declining (at least in France). Today we note a tremendous growth in the evidence bearing on the perception and observation of the other, the neural bases of emotions, etc.: the solipsistic brain has given way to the social brain. The new fast-growing interest in so-called neuro-economy and the development of experiments in cerebral imagery bearing on empathy and the perception of emotions in the other attest to all this.
The Politics of Prognostication: Astrology, Political Conspiracy and Murder in Fifteenth-Century Milan
Authors: Azzolini Monica
Published: November 2023

On 26 December 1476 Galeazzo Maria Sforza, second duke of Milan, was brutally assassinated while attending mass in the Church of Santo Stefano in Brolio. The three culprits were three young Milanese patricians, Giovanni Andrea Lampugnani, Carlo Visconti, and Gerolamo Olgiati. The basic events that preceded and followed the assassination are fairly well known: Galeazzo’s tyrannical style and rather volatile temper had not endeared him to the Milanese nobility (Fig. 1).
Accessing nature, circulating knowledge: Conrad Gessner’s correspondence networks and his medical and naturalist practices
Authors: Delisle Candice
Published: November 2023

Conrad Gessner led a busy life. Born in 1516, he had produced when he died aged 49, fifty-nine books on a variety of subjects. He left hundreds of paintings of plants, and enough material for more than ten substantial publications. In 1553, he had been elevated to the position of first town physician for the Canton of Zurich. He therefore had to attend poor patients, to survey and visit the hospitals, to preside over the Wundgschau and the diagnoses of leprosy for the whole Canton, to assess and provide help to midwives, surgeons, and apothecaries & and to lecture every day at the Zurich’s Grossmu¨nster school, the local high school, on physics.
‘The loving labours of a learned German’: Adolf Michaelis and the historiography of classical sculpture in Britain
Authors: Coltman Viccy
Published: November 2023

In a letter of 20 September 1877, Adolf Michaelis, Professor of Classical Archaeology at the new Kaiser-Wilhelm-Universitaät in Strasbourg, wrote from the London home of George Scharf, Director of the National Portrait Gallery, to the Right Honourable W. Cowper Temple at Broadlands in Hamp-shire: ‘beg[ging] your pardon for having delayed so longtime the returning the Memorandum [figure 3] and sending my slight Notes on your Collection [figure 4]’. The German academic had compiled these notes during his third research visit to England, undertaking exhaustive, first-hand, study for a forthcoming publication devoted to ancient sculptures in English private collections. Michaelis explained all this in a letter dated 25 August 1877 to the 5th Marquis of Lansdowne, whose ‘matchless collection’ of ancient sculptures at Lansdowne House in Berkeley Square, London he had previously examined during his second visit to Britain in 1873 and to which he wished to gain access again ‘to make notes upon the marbles, in order to give a fuller account of them’.
Bumblebees Abroad: Effects of Introduced Bees
Authors: Goulson Dave
Published: November 2023

The devastating impacts which some exotic organisms have wreaked on native ecosystems are all too familiar, and surely ought to have taught us a lesson as to the perils of allowing release of alien species. The introduction of Nile perch to Lake Victoria, of cane toads, prickly pear, rabbits, foxes and cats amongst numerous others to Australia, and of water hyacinth, Eichhornia crassipes , to waterways throughout the old world tropics, are perhaps some of the best known examples, but they represent only the tip of the iceberg. Australia alone had 24 introduced mammal species, 26 birds, 6 reptiles, 1 amphibian, 31 fish, more than 200 known invertebrates and no less than 2,700 nonnative plants at the last count (Alexander 1996; reviewed in Low 1999).
The Chronicle of Morea
Authors: Shawcross Teresa
Published: November 2023

The Chronicle of Morea, one of the most important and controversial historical narratives written in the late Middle Ages, tells the story of the formation and government by the Villehardouin dynasty of a remarkably successful Crusader State following the conquest by western invaders of the capital - Constantinople - and the provinces of the Byzantine Empire. By examining all the Chronicle's surviving Greek, French, Spanish and Italian versions, this study, the first of its kind, explores in depth the literary and ideological contexts in which the work was composed, transmitted and re-written. The result is a fascinating analysis of cultural exchange in a rich and vibrant eastern Mediterranean world where different ethnicities were obliged to live alongside each other, and outside political interests frequently intruded in dramatic fashion. Translations into English have been provided of all the material discussed.
The Human History
Authors: Gerald O’Collins S J
Published: November 2023

He [Jesus] was conscious of a vocation from God to proclaim this kingdom, and the record shows him as single-mindedly devoted to that vocation, even to the point at which it brought him to death. (John Macquarrie, Jesus Christ in Modern Thought) ‘ The Word became Mesh and made his home among us’ (John 1: 14). We might gloss the climax of John’ s prologue by saying that ‘ the Word took on a human history’ . Chapter 1 has already indicated certain issues connected with our historical knowledge of Jesus.
Lord, Saviour, God, and Spirit
Authors: Gerald O’Collins S J
Published: November 2023

It was their [the Christians’ ] habit on a fixed day to assemble before daylight and recite by turns a form of words to Christ as God. (Pliny the Younger, Letter 10) Those who wish to acquire a more adequate sense of how New Testament Christians evaluated Jesus’ being and doing rightly examine other high christological titles like ‘ Lord’ , ‘ Saviour’ , and ‘ God’ . Such titles exemplify further what Paul, John, and other New Testament witnesses held to be important about Jesus.
Medieval and Modern Christology
Authors: Gerald O’Collins S J
Published: November 2023

The ovens is cancelled only by love. (Thomas Aquinas, Contra gentiles) Any account of medieval Christology cannot ignore three lines of development, which may be labelled academic, monastic, and popular. Anselm’ s younger contemporary Peter Abelard (1079–1142) had an important impact on the scholastic method of debate to be used in the emerging European universities and other theological centres.
Universal Redeemer
Authors: Gerald O’Collins S J
Published: November 2023

In a real sense, only one human being will be saved: Christ, the head and living summary of humanity. (Pierre Teilhard de Chardin, Le Milieu divin) That which is called the Christian religion existed among the ancients . . . from the beginning of the human race until Christ came in the fresh, at which time the true religion which already existed began to be called Christianity. (Augustine, Retractationes) From its earliest to its later books, the New Testament does not waver in acknowledging Christ as the one Saviour for all people.
Introduction
Authors: Perry Curtis , Watkins John
Published: November 2023

Shakespeare played an incalculable role in shaping later impressions of the Middle Ages. Almost any book written on the Hundred Years War or the Wars of the Roses begins by explaining just how Shakespeare got it wrong. He conflated characters, condensed chronologies, cleaned up some careers, and sullied others. He turned Joan of Arc into a sexually voracious sorceress and suppressed Humphrey of Gloucester’s ambitions by characterizing him as a wise, disinterested councilor. His views of the conflict between York and Lancaster could not have been more biased. Even though we no longer see them as a reflection of Tillyard’s Elizabethan World Picture, we know that they derived more from sixteenth-century concerns about rebellion against the highly organized Tudor state than from anything like a modern scholar’s understanding of the parcellized sovereignty of late medieval England. If you want to know something about what actually happened during the fourteenth and fifteenth centuries, Shakespeare is not the best place to start.
Shakespeare’s Fickle Fee-Simple
Authors: Warley Christopher
Published: November 2023

The first adjective used to describe the central character in Shakespeare’s A Lover’s Complaint—a maid who has been seduced by a young man—is ‘fickle’, and ‘fickle’ seems a pretty good description of the poem generally. Everything in the poem appears changeable, inconstant, unreliable, and perhaps—in an older sense of the word—false, deceitful, and treacherous. This mutability of the poem has generally been taken as a bad thing: readers have tended to find it hard going, and criticism has by and large not been kind. The work has regularly been seen (apologetically) as unfinished or (optimistically) as not by Shakespeare. Whatever one thinks about such arguments, the shiftiness and difficulty—call it the ‘transitoriness’—of the work remain conspicuous. Writing in 1922, J. Denton Snider registered the confusion many readers have felt: the poem, he lamented, ‘employs no Greek Mythos (like Venus and Adonis), no Roman Tale (like Lucrece) for its scaffolding’.3 Without a scaffold, A Lover’s  Complaint seems simply to fall down.
Helping problem drug users
Authors: Robson Philip
Published: November 2023

The essential first step towards obtaining help with a drug-related problem (throughout this chapter ‘drug’ will include alcohol and tobacco) is to recognize that one exists. The affected person is often the last to realize or admit that things are getting out of hand, and it falls to friends or family to bring it to his or her attention.
International drug policy—is it working?
Authors: Robson Philip
Published: November 2023

The 182 current signatory nations to the Single Convention on Narcotic Drugs (1961), the Convention on Psychotropic Substances (1971), and the Convention against Illicit Traffic in Narcotic Drugs and Psychotropic Substances (1988) are required to ‘limit to medical and scientific purposes the cultivation, production, manufacture, export, import, distribution of, trade in, use, and possession of [certain] drugs’. Draconian enforcement of controls is imposed because of the grave public health, social, and economic risks the drugs are said to pose.
Biology is only part of the story
Authors: Read Dwight , Leeuw Sander van der
Published: November 2023

In this paper, we are interested in extrapolating from what we know about the development of prehistoric technologies to the evolution of human cognition. Our interest is driven by the following (unsolved) questions concerning the history of human technology: (i) Given that primates, for millions of years, have made and used simple tools, but have not developed any complex technologies, what is different about human beings that enabled them to develop the latter? (ii) Are the enabling factors biological, are they social cultural or are they a bit of both?(iii) When was that capacity acquired? Was that the result of a sudden change, an incremental one or a more complex process that combined both? (iv) Why did it take so long to ‘invent’ and accelerate innovation? (v) Why did the rate of innovation increase so rapidly, once that point was reached? Numerous authors (e.g. Stout et al. 2000 and references therein) have argued that changes in the technology of artefact production and in artefact form, variety and diversity relate to cognitive changes, but what constitutes those cognitive changes has remained unclear due to our as yet imprecise understanding of how the brain functions.
Talking Science at the University of Padua in the Age of Antonio Vallisneri
Authors: Dooley Brendan
Published: November 2023

Even for an accomplished scholar like Antonio Vallisneri, it was no simple matter to declaim extemporaneously in Latin before a public possibly demanding the best from a setting where Vesalius and Fabricius had once changed medical education forever in the West. Therefore successful completion of the inaugural lecture was a cause for celebration, so he wrote to the Tuscan grand ducal librarian Antonio Magliabechi in 1700: ‘Today is a holy day for me, since I made my solemn entrance into the university favoured by the applause of all the learned, the podesta`[of Padua] and the captain of the city’.
Microbial biosensors
Authors: Karube Isao , Suzuki MasaYasu
Published: November 2023

Recently, many kinds of biosensors have been developed for the determination of organic compounds (1-3). Enzyme sensors are highly specific for substrates of interest, but the enzymes employed are generally expensive and unstable. Microbial sensors are composed of immobilized microorganisms and an electrochemical device and are suitable for on-line control of biochemical processes.
Viral–receptor binding assays
Authors: Schwimmbeck Peter L , Löhr Matthias , Tishon Antoinette
Published: November 2023

Viral receptors are the main determinants for viral tropism. Receptors govern pathogenesis by allowing a cell to be susceptible by its attachment to virus. The primary function of receptor proteins on cells is, of course, not the promotion of an infection but a function in the life cycle of the cell. Viruses may utilize several specific host surface proteins like CD4 (human immunodeficiency virus), the Cr2 component of C3 (Epstein Barr Virus), ICAM-1 (rhinovirus), a new member of the immunoglobulin superfamily (poliomyelitisvirus), the adrenergic-like receptor (reovirus) or several proteins of defined molecular weights but not functionally defined (coronaviruses, lymphocytic choriomeningitis virus, etc). Restricted presence of the receptor molecules may account for the tissue specificity along with virus-specific enhancer sequences (1-4). In addition, unrelated viruses may share receptors on a given cell (5).
Viral-autoimmune experimental models
Authors: Fujinami Robert S
Published: November 2023

The approach taken for the analysis of virus induced autoimmune disease has been adapted from that for the examination of experimental autoimmune diseases such as allergic encephalomyelitis. The major principle is to use defined viral peptides sharing common amino acids with disease inducing regions from self proteins in order to initiate disease (1). Thus, critical viral determinants which share similar regions with host disease inducing epitopes need to be identified. For our consideration here we will confine the descriptions to the central nervous system (CNS) autoimmune disease and experimental allergic encephalomyelitis (EAE). However, similar approaches can be taken for the study of other organspecific experimental autoimmune disease such as thyroiditis, diabetes, orchitis and myocarditis.
Dissociated neurons from adult rat hippocampus
Authors: Chad J E , Stanford I , Wheal H V , Williamson, R , Hall G Wood
Published: November 2023

For quantitative experiments on neurons it is imperative that the number of variables be minimized. Ideally the voltage across the whole-cell membrane and the ionic conditions on both sides of the membrane would be controlled. This ideal is most nearly achieved with isolated membrane patches (1, 2), but this also has the effect of removing the influence of the cell metabolism. A useful compromise between control and integrity of the cell has been the perfusion and voltage clamping of isolated spherical neurons (3, 4). Under these conditions voltage and drug activated currents can be accurately determined and their modulation by intra- or extracellularly applied agents measured.
Chemical synapses in cell culture
Authors: Haydonandm P G , Zoran .J
Published: November 2023

A topic of considerable interest and debate since the discovery of the ability of the vagus nerve to release acetylcholine (ACh) has been the investigation of the mechanisms of synaptic transmission. Such studies are hampered. however, due to the small size of most synaptic terminals and due to the inaccessibility of neurons in vivo. The elucidation of cellular and molecular mechanisms of synaptic development and transmission has been facilitated in recent years by techniques which permit the reconstruction of specific neuronal elements in cell culture. By plating neurons in cell culture it is possible to study specific synapses in isolation. For example, with a single pair of neurons in cell culture it becomes possible to monitor spontaneous transmitter release from a defined pre-synaptic cell, an approach that is impossible in vivo due to the polyneuronal innervation of most neurons.
Annexins: Calcium-Dependent Phospholipid-Binding Proteins
Authors: Boustead Catherine M , Walker John H
Published: November 2023

1. Introduction The effects of calcium as an intracellular second messenger are mediated by its interaction with calcium-binding proteins. Recently, a new family of calcium-binding proteins, the annexins, has been identified in a wide variety of species and tissues (for reviews see refs 1-4). Annexins are abundant proteins (up to 0.5% of cell protein) which bind reversibly to phospholipids at micromolar calcium concentrations, and inhibit the action of phospholipase A2 in vitro. Certain annexins are also substrates for serine and tyrosine kinases, and some exhibit calcium-dependent binding to F-actin. Structurally, the annexins are characterized by a variable N-terminal tail, followed by four (in the case of annexins I-V and VII-VIII) or eight (annexin VI) repeats of a highly conserved 70 amino acid sequence.
Site-directed mutagenesis using gapped-heteroduplex plasmid DNA
Authors: Inouye Sumiko , Inouye Masayori
Published: November 2023

Oligonucleotide-directed site-specific mutagenesis is the most effective and powerful method to create desired mutations at a specific site in a gene. The method involves the use of a short, synthetic oligonucleotide carrying the appropriate mutation, which acts as a primer of in vitro DNA synthesis on a complementary single-stranded circular DNA template. There are two basic methods to create a complementary single-stranded circular DNA template; one utilizes double-stranded plasmid (the plasmid method) and the other utilizes single-stranded M13 or phagemid vectors (the M13 method). The M13 method originally developed by Zoller and Smith (1, 2) has been modified to obtain higher yields of mutants (see Chapters 1 to 3). Today, kits for the M13 method are commercially available with which one may get desired mutations with a yield of higher than 50%. However, one of the drawbacks of this method is that one usually has to redone a target fragment into an M13 vector for mutagenesis. In addition, one has to move the mutated fragment back to the original expression plasmid after mutagenesis. In contrast, the plasmid method allows one to directly use double-stranded plasmid DNA for mutagenesis. Although the yields of a desired mutation are usually lower (3-25%) with the plasmid method than with the M13 method, in practice the yields with the plasmid method are high enough to easily obtain desired mutation by a simple screening method.
Fixation
Authors: Hopwood David , Milne Gordon
Published: November 2023

The techniques chosen to fix tissues should enhance the sort of microscopical information which is eventually produced. Ideally, the tissue should be retained in as near life-life condition as possible, but the better the morphological preservation, the less the biological activity and vice versa. Some compromise has to be reached. Fixation should also prepare tissues to withstand the subsequent rigours of the preparative techniques (Chapter 2) and finally irradiation by the electron beam (1, 2).
Freeze-substitution and other low temperature embedding methods
Authors: Roos Norbert
Published: November 2023

The typical living cell is an extremely complex and usually dynamic entity. It is surrounded by aqueous domains and contains a variable amount (usually more than 70%) of non-uniformly distributed water, which is an important structural component of most biological systems.
Quantitative biological X-ray microanalysis
Authors: Warley Alice , Gupta Brij L
Published: November 2023

When a specimen is examined in the electron microscope the interaction of the electron beam with the atoms of the specimen results in the generation of X-rnys: these originate from two different sources. The interaction of the electron beam with orbiting electrons in atoms of the specimen causes the emission of characteristic X-rays, whose energy depends on the atomic number of the element in which the transition is occurring and the electron shells involved.
Crystallization of Nucleic Acids and Co-Crystallization of Proteins and Nucleic Acids
Authors: Dock-Bregeon A -C , Moras D
Published: November 2023

At a first glance crystallizing nucleic acids poses the same problems as crystallizing proteins since the variables under investigation arc alike. Some differences can be underlined, however. Most of the papers dealing with crystallization of biological macromolecules describe protein crystal growth. Thus it seems important to add a contribution for nucleic acid crystallization. As seen from crystallization data banks and from the Protein Data Rase, it is obvious that these banks contain a large majority of proteins; this is largely due to the difficulty in obtaining large single crystals of nucleic acids.
Isolation of intercellular junctions
Authors: Hertzberg Elliot L , Stevenson Bruce R , Green, Kathleen J , Tsukita Shoichiro
Published: November 2023

Early electron microscopic studies revealed the existence of a number of distinct types of intercellular junctions. Specific physiological properties of a cell or tissue could often be correlated with the presence of a particular type of junction, suggesting very different functions for each type. Some types of cell contacts, the desmosomes and adherens junctions, appeared to tether different elements of the cell’s cytoskeleton to the plasma membrane while forming adhesion points between cells. Others were proposed to provide a barrier to paracellular diffusion of solutes (tight junctions) or facilitate intercellular communication (gap junctions). Roles were hypothesized for these junctions in tissue modelling, and in regulation of cell function and cell proliferation.
Multiple regression
Authors: Iles Terence c.
Published: November 2023

The previous chapter covers methods of fitting equations to data where there is just one predictor variable x for the dependent variable y. Sometimes data are collected on two or more different predictor variables x, x, … xk and an equation is sought to calculate y from the set of measurements of x  x, … x  k. This chapter describes the fitting and interpretation of linear equations with such data sets. The choice of the variables that should be included in the prediction equation, and the possible transformation of variables, will also be discussed. The calculations required for multiple regression are usually too time-consuming to be done by hand so it is necessary to use a computer. Fortunately, most statistical packages have regression routines and these give the user a wide choice of statistics that help in finding suitable prediction equations. Many of these diagnostic statistics are most easily interpreted by the use of plots, so a package with built-in plotting routines should be used.
Ordination
Authors: Randerson P F
Published: November 2023

Individuals comprising a population, or at least a sample taken from it, may be described quantitatively and compared by measurements of common characters or variables. Thus a variable is any quantity that takes different values for different individuals or for the same individual at different times, that is a quantity which can take any one of a specified set of values.
Dynamic models of homogeneous systems
Authors: Bowker David w.
Published: November 2023

This chapter provides instructions for the development of simple mathematical models which simulate the dynamics of single species populations of organisms in cultures and natural environments. These models are particularly suitable for biologists with limited previous experience in modelling because they are conceptually straightforward, mathematically tractable, and easy to implement on a computer. They consist of equations which recur frequently in modelling to describe fundamental processes such as exponential growth and decay, and they serve as a starting point from which the beginner can acquire sufficient expertise to progress to more complex modelling problems such as those described in Chapter 9.
Toxin ablation in Drosophila
Authors: Steller Hermann
Published: November 2023

Methods for the selective removal of specific cells have been widely used to investigate the influence of a particular cell on the development of its neighbours. In the past, cell ablation techniques have primarily relied on physical methods, including classical surgical techniques that date back to the origin of developmental biology (1). More recently, laser microbeam ablation has been used extensively for developmental and functional studies in the nematode Caenorhabditis elegans (2, 3), and also to investigate certain aspects of insect development (4, 5). Unfortunately, the small size and opaque cuticle of Drosophila place severe restrictions on the widespread use of laser ablation in this organism. In addition, physical ablation methods are only practical for killing rather small numbers of cells, but fail when it is necessary to remove larger cell populations, in particular if its members are distributed widely throughout the animal. Finally, given the Jabour-intensive nature of the technique, it is usually not possible to produce a sufficient number of individuals needed for genetic, biochemical, or behavioural analyses.
Introduction to cell cycle controls
Authors: Hartwell Leland
Published: November 2023

A theory for eukaryotic cell cycle control has emerged in the last few years that has achieved a consensus in the research community. Although many surprises undoubtedly await the detailed testing of this theory, it is likely that its major tenets will stand the test of time. In this introductory chapter I will present a broad outline of our current understanding of how the cell cycle is controlled and raise a number of questions that are presently motivating cell cycle research. Most of the topics discussed in this chapter have been extensively reviewed recently; consequently, where possible, I will refer the reader to these reviews for documentation of the primary literature.
Start and the Gl-S phase transition in budding yeast
Authors: Reed Steven I
Published: November 2023

Mechanisms to ensure that division occurs in a coordinated and regulated fashion are prerequisite for fitness and ultimately for viability of all cells. This means that the division process must be responsive to a host of internal and external signals that mediate, among other things, integration of division with the extracellular milieu, coordination of division with cellular growth and the occurrence of cell cycle events in the proper order. In the budding yeast, Saccharomyces cerevisiae,both the regulation of division by external signals and the coordination of growth and division occur in the Gl interval of the cell cycle (1). Operationally, there are two types of signal that severely restrain division in Gl in this organism: nutritional limitations (2–13) and the action of mating hormones (4, 14-22). Although the regulatory intents of these signals are quite disparate, each causes a cell cycle arrest characterized by completion of the ongoing cell cycle and arrest in the subsequent Gl interval (1). Additionally, changes in the cellular growth rate are coordinated with the division process to ensure a relatively constant cell size by altering specifically the length of the Gl interval: slow growth is accommodated by extending the length of Gl while rapid growth shortens Gl (5, 23–38). These observations led to the hypothesis of a universal Gl control mechanism in yeast (1). In its simplest formulation, this hypothesis states that all signals governing passage through Gl and entry in S phase do so by regulating the completion of an essential Gl function or event designated START; completion of START commits a cell to S phase and division; delay of completion of START delays entry into S phase; failure to complete START prevents initiation of S phase and imposes alternative fates upon cells.
Regulation of the cell cycle during Drosophiladeveloptnent
Authors: White-Cooper Helen , Glover David
Published: November 2023

It is perhaps surprising that relatively little is known about the regulation of the cell cycle during development, particularly as many of the organisms now used to study the cell cycle were originally the focus of studies of both cell division and development in the late nineteenth century. The classical studies that defined mitosis and its phases, together with the cyclical behaviour of chromosomes and centrosomes, originated from observations made upon developing embryos of a variety of organisms. The nineteenth century view of the cell cycle was one of alternating periods of cell division and a vegetative state. Not until the 1950s did the studies of Howard and Pelc (1) define a discrete period of DNA synthesis (S phase) within this vegetative phase, thus leading to the archetypal view of the cell cycle in which mitosis (M) is separated from S by two gap phases Gl and G2. There are, however, many variations on this theme that are particularly evident during development. Many embryos undertake a series of rapid cell cycles during their early development driven by maternally provided proteins. The Drosophilaembryo begins its existence as a syncytium that becomes populated with several thousand nuclei as a result of 13 extremely rapid rounds of synchronous mitosis at approximately 10 min intervals. These mitoses are separated by periods of DNA synthesis without intervening gap phases. A G2 phase is introduced into the cell cycle only after individual nuclei have become incorporated into cells during division cycle 14. A Gl phase is not introduced into the Drosophilacell cycle until the seventeenth division, and only then do the cells of the embryo undertake ‘conventional’ cell cycles. These occur in the CNS and imaginal tissues, which continue to proliferate throughout larval and pupal development. In most other tissues the cell cycle is far from conventional, since Drosophila,in common with other dipterans, has many tissues whose cells undergo endoreduplication cycles.
Eukaryotic mRNA Processing
Authors: Krainer Adrian R
Published: November 2023

This volume focuses on the major aspects of post-transcriptional mRNA processing in the nucleus of eukaryotic cells. Each of the described mRNA reactions is required for proper gene expression and can also serve as a control point for regulating the expression of many genes, for example during embryonic development or in different cell types. The different chapters review the assembly of newly synthesized nuclear mRNA transcripts into hnRNP particles and catalytically active spliceosomes; the structure and mechanism of action of small nuclear ribonucleoprotein particles and protein factors that catalyse pre-mRNA splicing in mammalian cells and in yeast; the regulation of gene expression and generation of protein isoform diversity by alternative splicing; the mechanisms of 3' end cleavage and polyadenylation; the architecture of the cell nucleus in relation to these processes and to the localization of the relevant substrates and factors; the diverse mechanisms of RNA processing by ribozymes and their potential relevance for nuclear mRNA processing; the mechanism of spliced-leader addition by trans-splicing in nematodes and trypanosomes; and the process of insertion/deletion mRNA editing in kinetoplasmid protozoa. In each chapter, leading researchers have provided detailed, critical reviews of the history, experimental approaches, major advances, current ideas and models, as well as future directions, for each of these active areas of research.
Postimplantation mammalian embryos
Authors: Morriss-kay Gillian m.
Published: November 2023

Techniques for culturing whole postimplantation mammalian embryos were pioneered by D.A.T. New in Cambridge (1, 2). Originally, rat embryos were cultured on plasma clots in watch glasses. Better development was found to result from the use of blood serum as a culture medium and by the use of ‘circulators’, in which continuously gassed serum passes as a constant stream through a chamber containing the embryos. The serum was obtained after allowing blood to stand and clot; embryonic development was quite good in that somites formed and the trunk elongated, but the two heart tubes often failed to fuse, and the trunk was excessively rigid, often protruding out from the curve of the yolk sac. These problems were overcome by centrifugation of the blood immediately after removal from the animal, so that the clot did not form in contact with blood cells (3). The serum is also heat-inactivated (30 min at 50 °C) to remove complement.
Cell Transplantation in Xenopus
Authors: Altaba Ariel Ruiz I
Published: November 2023

Transplantation of cells in Xenopus embryos can be easily achieved. Early embryonic cells, blastomeres, can be separated, dissociated, and cultured with ease due to their large size and the fact that all cells contain a storage of yolk. This chapter describes simple procedures for transplanting single labelled blastomeres and for grafting small groups of cells into host embryos.
RNAase protection assays
Authors: Altaba Ariel Ruiz I
Published: November 2023

The development of a sensitive assay with which reliably to measure and map messenger RNAs (1, 2) has proved very useful in the study of the molecular mechanisms underlying inductive and differentiation events. The transcription of specific genes is often taken as a reliable marker of the commitment of the expressing cell towards one fate or another. For example, embryonic cells expressing a-actin are thought of as cells differentiating along a muscle pathway and those expressing N-CAM along a neural pathway. Early regulatory genes, the expression of which may be crucial in the commitment of a cell to a given fate, are also used as markers, although the expression of these genes should always be interpreted cautiously. Here I describe a method, based on that of Krieg and Melton (3), to measure the levels of mRNAs in embryonic cells by RNAase digestion of RNA:RNA hybrids. For general molecular techniques the reader is advised to consult ref. 4.
Construction of representative genomic DNA libraries using phage lambda replacement vectors
Authors: Kaiser Kim , Murray Noreen E , Whittaker Paul A
Published: November 2023

In the years since this article first appeared, the technology it describes has become much more routine. This is in large part due to the contribution that the biotechnology industry has made to the provision of reliable reagents, ready-prepared vector DNA (cleaved and dephosphorylated if necessary), packaging extracts, host strains, and even kits to take one through the entire procedure from beginning to end. Failing all else one can send away a tissue sample and receive, if at some cost, a library in the post.
Procedures for cDNA cloning
Authors: Watson Christine J , Demmer Jerome
Published: November 2023

The first mammalian cDNA was cloned almost two decades ago (1) and the construction and screening of cDNA libraries has since become one of the fundamental procedures of molecular biology. A considerable number of advances have been made during the past 20 years, including the availability of highly purified, cloned, and modified enzymes which has improved the efficiency of cDNA synthesis and cloning. The development of high efficiency cloning systems has allowed the construction of a representative cDNA library to become a straightforward procedure which can be successfully carried out in any molecular biology laboratory. More recently, the polymerase chain reaction (PCR) has revolutionized cDNA cloning and in some circumstances it will not be necessary to construct a library for the isolation of a particular cDNA clone.