bluuebunny/crossref_metadata_2025 huggingface dataset:
The Level of Decision Making: The Preferences of the Citizens after Enlargement
Authors: Winter Lieven De , Swyngedouw Marc , Goeminne Bart
Published: November 2023

In the current system of multilevel governance in the EU, different policy issues and areas are dealt with at various levels (ranging from the local to the EU), sometimes exclusively at one level, but often by joint decision making between two or more levels. The question of whether or not the level(s) at which decisions are currently taken in different policy fields is truly the most appropriate level is one of the aspects that is supposed to affect the legitimacy and effectiveness of any democratic polity (Thomassen and Schmitt 1999a; Peterson 1995).
chapter 2
Authors: Wu Duncan
Published: November 2023

London was endlessly stimulating. Young William fell in love with its restlessness, often wandering out of the front door and rambling about on his own. His parents were terrified he would get lost but he seemed always to find his way home. Hazlitt Sr occasionally walked him to the Montpelier Tea Gardens in Walworth, planted with trees, shrubs, and exotic flowers. These were the first green things the boy had seen after the long sea voyage back to England and he was fascinated by them—their names, colours, and odours.
political theory and the realistic spirit
Authors: Emmerich Ze’ev
Published: November 2023

I open books on right and on ethics, I listen to the scholars and jurisconsults and, moved by their ingratiating discourses, I deplore the miseries of nature, I admire the peace and justice established by civil order, I bless the wisdom of public institutions, and console myself for being a man by seeing that I am a citizen. Fully instructed about my duties and happiness, I close the book, leave the class-room, and look around me; I see unfortunate peoples groaning under an iron yoke, mankind crushed by a handful of oppressors, starving masses overwhelmed by pain and hunger, whose blood and tears the rich drink in peace, and everywhere the strong armed against the weak with the frightful power of the laws.
Introduction
Authors: Saggar Shamit
Published: November 2023

Reputations matter. At present, and for some while past, the reputation of Muslims in Western societies, regrettably, is all but in shreds. Though this statement may strike some as harsh, it is no exaggeration to suggest that all-around hardly anyone has anything remotely good to say about Western Muslim communities. Meanwhile, the reputation of Western liberal societies in the eyes of even moderate Muslim opinion is little better than in the gutter. The essential elements of a siege mentality have set in, and are founded on the widespread pariah reputation attached to Western Muslims.
Concluding Thoughts and Future Prospects
Authors: Saggar Shamit
Published: November 2023

Considerable space and attention has previously been devoted in this book to diagnosing aspects of the problem or problems at hand. Indeed, it is striking that the shelves of volumes that have been published on this and similar topics have become heavily congested with detailed and authoritative diagnoses of one kind of another. In the case of Western Muslims and issues of integration and security, there is no mistaking the fact that publisher, reader, and even artistic interest, has swollen massively. A mighty avalanche has been seen in the past half-dozen years, mainly involving either snappy commentaries on particular controversies or noble accounts of specific pieces of evidence and their meaning. In both cases, the emphasis on analysing and dissecting the problem itself has been very sharp and has been at the expense, generally speaking, of equal attention being paid to the thorny question of plausible solutions and responses, to say nothing of the effectiveness of current interventions.
How to Do Science in Papua New Guinea
Authors: Novotny Vojtech
Published: November 2023

Scientific progress in New Guinea was hampered by the fact that the people traditionally lived in tiny populations, isolated by language, and so by lack of access to information; and they were also illiterate, which meant that all knowledge was passed down from generation to generation by word of mouth. It is unsurprising that their technical advancement was slow and that they possessed only a small body of scientific knowledge. Science as understood in the West— the obsessive accumulation of all manner of facts, including the totally useless—is a novelty in Papua New Guinea and is still looking for a wider circle of acolytes. But the natural and cultural riches of the island have always been an irresistible lure to Western biologists and anthropologists. Huge numbers of these continue to criss-cross the country, spreading their scientistic faith and adding greatly to the local colour.
In Conclusion: Travel Report
Authors: Novotny Vojtech
Published: November 2023

When my colleague William Boen and I set off on a routine inspection of our outlying workplace, the entomological laboratory in the village of Mu, Simbu province, in the Highlands of New Guinea, we used public transport as usual. The generally smooth journey through the jagged Finisterre Range was enlivened only by our having to get out and push the twelve-seater minibus up some of the particularly steep hills. We also had to ford one river to get round a bridge that was hopelessly blocked by a lorry. The lorry had sunk through the planking of the single-lane bridge; the next day it was hanging by its chassis on the steel frame of the bridge.
Voice for the Weak: ECOSOC Ad Hoc Advisory Groups on African Countries Emerging from Conflict
Authors: Prantl Jochen
Published: November 2023

In 2002, the United Nations created networked groups to strengthen the coherence of the organization’s relations with countries emerging from conflict. The ‘ad hoc advisory groups’ were created by the Economic and Social Council of the United Nations—the principal organ for coordinating the economic, social, and related work of the 14 UN specialized agencies, functional commissions, and five regional commissions. This chapter gives an assessment of the role and performance of the ECOSOC ad hoc advisory groups for African countries emerging from conflict. It examines the potential and limits of these groups in providing conflict countries and other stakeholders with a greater voice in the transition process from war to peace. In this context, ‘voice’ includes the attainment of status and the mobilization of international (financial) support, as well as influencing decision-making at the UN Headquarters and the international financial institutions. ECOSOC ad hoc advisory groups are set up at the request of any African country emerging from conflict and seek to facilitate the integration of relief, rehabilitation, reconstruction, and development into a comprehensive approach to peace and stability.
The Heavily Indebted Poor Countries’ Finance Ministers’ Network
Authors: Martin Matthew
Published: November 2023

The Heavily Indebted Poor Countries’ (HIPCs’) Finance Ministers’ Network was established in 1999 as a response by Finance Ministers of the HIPCs to the creation of the HIPC Debt Relief Initiative in 1996. It stemmed from the actions of a wider technical capacity-building programme (the HIPC Debt Strategy and Analysis Capacity Building Programme) in which they had come to have a considerable degree of trust since its establishment in 1997. Their growing dissatisfaction with the construction and progress of the original HIPC Initiative led them to request the creation of a forum where they could have frank discussions among themselves (and sometimes with representatives of like-minded donors and the Bretton Woods Institutions, or BWIs) about prospects for debt relief. Over time, the network also became a forum for discussing poverty reduction strategies, new development financing prospects, and their capacity-building needs, and its membership has grown from 16 to 34 countries. The network has fostered learning among HIPC Finance Ministers and their officials, by allowing them to exchange information and views on the progress of their debt relief, poverty reduction, and new financing. It has also served as a mechanism for HIPC Finance Ministers better to exercise voice in the international system, to access enhanced debt relief and better quality new financing, and to decide autonomously their collective positions on international debt and development financing issues.
Conclusions and Recommendations
Authors:   ADB
Published: November 2023

Oil and gas resource endowments are a source of wealth and can be expected to support growth and development if well managed. This involves converting the wealth into physical and human capital, increasing and sustaining economic growth, and alleviating poverty. This report has clearly outlined that the successful pathway to fully harnessing the benefits of oil and gas resource wealth in Africa is nested in good institutions and governance, trade links, sound economic policies, and heavy investments in exploration technology, legislative framework, and human capital development. Historically, oil and gas resource endowments in Africa have not yielded positive economic and development results for many countries. The experience has been more positive recently, and there are encouraging signs of better management and governance.
Blindsight
Authors: Weiskrantz Lawrence
Published: November 2023

Blindsight is an unusual condition where the sufferer can respond to visual stimuli, while lacking any conscious feeling of having seen the stimuli. It occurs after a particular form of brain injury. The first edition of 'Blindsight', by one of the pioneers in the field - Lawrence Weiskrantz, reported studies of a patient with this condition. It was an important, much cited publication. In the past twenty years, further work has been done in this area, and this new edition brings the book up to date. Retaining the original text, but adding substantial new chapters and colour illustrations, the first section of the book summarizes findings on DB since the last published account in 1986. The second part includes information on other new research that has occurred since the last edition. As well as giving an account of research over a number of years into a particular case of blindsight, it provides a discussion of the historical and neurological background, a review of cases reported by other investigators, and a number of theoretical and practical issues and implications. The book will be valuable for cognitive psychologists and cognitive neuroscientists, as well as philosophers of mind.
Background
Authors: Weiskrantz Lawrence
Published: November 2023

There is a paradox concerning the visual pathways and their functions. In man and other primates most of the fibres from the optic nerve leaving the eye project more or less directly to a region of the cerebral cortex situated at the most posterior region of the brain (Figs. 4.1 and 4.2), in a region known as the occipital lobe. The specific cortex within the occipital lobe which receives this visual input is called the ‘striate cortex’ because of its distinctive appearance under the microscope (also known as area 17, area OC, or V–1). Actually, the projection from eye to cortex is not absolutely direct: the optic nerve fibres themselves terminate in a nucleus of the thalamus in the ‘tweenbrain’ (the diencephalon), in the dorsal lateral geniculate nucleus, and from that nucleus new fibres originate that terminate in the striate cortex.
Visual acuity
Authors: Weiskrantz Lawrence
Published: November 2023

In our original report (Weiskrantz et al., 1974), we used an Ives apparatus that generates a sine-wave grating when light is passed through a pair of diffraction gratings (Moiré fringes; the apparatus is the same as that described more fully in Weiskrantz and Cowey 1963). Only one position in the scotoma was tested originally. The grating was visible in a circular aperture, 6° 39’ diameter, which was placed so that its centre was either 8° 48’ to the left or right of the fixation point (53 cm from the subject), depending on which visual half-field was being tested. Stimuli were presented singly, and D.B. was asked to report ‘lines’ or ‘no lines’, and the spatial frequency varied randomly from trial to trial so that a threshold determination could be made. Even though D.B. reported not only seeing no lines at all, but not even the brightly lit aperture, his performance varied systematically with the spatial frequency, and an acuity threshold could be determined. The results are summarized in Fig. 5.10 in Part III, where it can be seen that his minimal separable acuity in the impaired field at this position was a creditable 15.8 cycles/degree, as compared with 20 cycles/degree in the mirror symmetric region of the intact right half-field.
‘Waves’
Authors: Weiskrantz Lawrence
Published: November 2023

In testing in 1982 (cf., for example, the Chapter 13 tests on the optic disc within the scotoma), but especially in 1983, D.B. reported an impression of ‘waves’ in parts of his field defect, especially in a region between the fovea and about 30° eccentric in the lower quadrant. The region had been plotted in Fig. 13.1 in Chapter 13. In 1983 we recorded our impression that his ‘live’ field was livelier and slightly larger than before. The experience is of a kind unlike anything in normal visual experience, and for which precise words seem to be lacking. The ‘waves’ can have some sort of form. They can be straight or curved, or can even have ‘squareness’. They can also be ‘quick’, which is how he described one of the Aimark targets in a reaching experiment. ‘A quick movement; not a light’. The square was described as producing ‘a sharp movement’. He also described the square on another occasion as producing ‘a corner-shaped wave’. The waves could also be thinner or thicker, e.g., in the uncurved triangle vs. curved triangle discrimination, he said, ‘There is a movement. I said “straight” when the waves are thinner, and “curved” when they are thicker and quicker’. But D.B. insisted that he was not ‘seeing’. Interestingly, he mentioned in 1983 that he experiences these waves in everyday life. If, for example, there is a moving cat in his field defect, this sets up waves of the same sort that he experiences when having a migraine attack. He said that he cannot be sure, in such a situation, whether or not he is having a migraine attack until the cat emerges into his good visual field! The sensitivity would also seem to be quite high, and can be induced by his own movement. For example, he said that when he is walking at dusk, he is unable to see on his right side because of the darkness, but his left side of the field is intrusive because it contains waves.
Matching between Impaired and Intact Fields
Authors: Weiskrantz Lawrence
Published: November 2023

If, as the results of the double dissociation between form and detection suggest, the impaired field is qualitatively different from the normal field, the interesting question arises whether communication between the two fields is possible and, if so, is efficient. The way we have approached this question is to present stimuli to the two fields simultaneously and ask D.B. to make a ‘same-different’ judgement. Two groups of such tests have been carried out, in September 1975, and in April 1982.
Overview
Authors: Roff Derek A
Published: November 2023

Computer modeling is now an integral part of research into evolutionary biology. The advent of increased processing power in the personal computer, coupled with the availability of languages such as R, S-PLUS, Mathematica, Maple, Mathcad, and MATLAB, has ensured that the development and analysis of computer models of evolution is now within the capabilities of most graduate students. However, there are two hurdles that, in my experience, discourage students from making full use of the power of computer modeling. The first is the general problem of formulating the question in a manner that is amenable to programming and the second is its implementation using one of the aforementioned computer languages. This is because the learning curve of each of these languages is quite steep, unless one already has prior computing experience as an undergraduate.
Genetic Models
Authors: Roff Derek A
Published: November 2023

The “Fisherian” optimality approach determines the optimal combination of trait values but not the evolutionary trajectory nor, in general, the variation to be expected about the optimal combination. Invasibility analysis is also primarily concerned with the Evolutionarily Stable Strategy (ESS) values; though assuming clonal inheritance, it can give insight into the temporal evolution of trait values. Whether such analyses can be extended to organisms with other modes of inheritance is not clear, though the analysis presented in Scenario 5 of this chapter suggests that caution is warranted in applying the results of invasibility analysis to sexually reproducing organisms.
Fermentation modelling
Authors: Sinclair C G , Cantero D
Published: November 2023

The purpose of fermentation modelling is to bring order to the mass of data which results from a practical fermentation experiment and to express the results in a concise form which is intelligible to colleagues and to those who wish to make use of the results. One of the principle uses of a fermentation model is to design large-scale fermentation processes using data obtained from small-scale fermentations. As an example, let us take a simple aerobic yeast fermentation. Pure yeast culture is first transferred from a slope to a shake flask, grown in an orbital shaker for 12 h; the shake-flask contents are then aseptically added to a 5 litre laboratory fermenter. This is equipped with the usual agitator, air supply control equipment, probes for measuring temperature, pH, dissolved oxygen and, hopefully, off-gas analysis equipment. It will have been charged with a growth medium and be free of organisms prior to inoculation. A sample of the inoculum and of the medium will be taken initially (time t= 0), then, every 30 or 60 min, a sample of the fermenter contents will be taken for analysis.
Methods for the measurement of insulin secretion
Authors: Malaisse Willy J , Leclercq-Meyer, Viviane , Malaisse-Lagae Francine
Published: November 2023

Several methods are currently available to assess the secretory activity of the pancreatic B-cell either in vivo or in vitro.In vivo, changes in insulin release are often judged from the level of circulating insulin. This method does not provide information on the absolute secretory rate, and the plasma concentration of insulin may be affected by extrapancreatic factors affecting its clearance from the blood stream. The true secretory rate can be measured by following the progressive neutralization of injected anti-insulin antibodies. Experiments carried out in vivo do not, however, show conclusively whether the effect of a given agent results from a direct or indirect action upon the pancreatic B-cell.
Cell biology of secretion
Authors: Brocklehurst Keith W , Pollard Harvey B
Published: November 2023

The aim of this chapter is to describe various techniques that have proved invaluable in attempts to elucidate the cell biology of secretion through the use of the bovine adrenal medullary chromaffin cell as an example of a secretory cell. We will describe procedures for the isolation and culture of chromaffin cells, and then describe the use of such cells as a system in which to investigate the effects of various activators and inhibitors of secretion. The assay of catecholamines and ATP as measures of the extent of secretion will be described. A procedure will then be detailed for rendering cells permeable using the detergent digitonin, and the advantages of using such cells to study secretory mechanisms will be discussed.
Regulation of the biosynthesis and processing of polypeptide hormones
Authors: Hutton John C , Bailyes Elaine M , Rhodes, Christopher J , Guest Paul C
Published: November 2023

The majority of polypeptide hormones are stored in intracellular vesicles which are bounded by a phospholipid bilayer and are released into the extracellular medium by a process of regulated exocytosis. The simple morphology of the secretory granule belies its biochemical complexity, for not only is it a store for the peptide hormone but it also contains proteases which may be involved in the conversion of precursor forms of the stored peptide, enzymes such as proton pumps involved in maintenance of the pH and ionic composition of the granule interior, and the granule membrane proteins which are involved in the mechanism and regulation of exocytosis. The protein composition of secretory granules from different endocrine tissues is poorly documented and the process of secretory granule biogenesis through which the granule derives its full complement of functional elements is only just beginning to be unravelled. The following techniques reflect our experience of studies on the biosynthesis and storage of insulin and the accessory proteins of the insulin secretory granule of the pancreatic ,8-cell. The protocols given should, however, be more widely applicable to the study of other endocrine tissues and cell lines transfected with polypeptide hormone mRNAs from heterologous sources.
Dissection and culture of postimplantation Embryos
Authors: Cockroft David l.
Published: November 2023

The inaccessibility of the postimplantation mammalian embryo in utero has stimulated the development of a number of techniques to facilitate its manipulation. The culture methods described in this chapter allow great control over the embryos’ environment and afford good access for injection or surgical procedures. The major disadvantage of such in vitro techniques is the relatively short period (usually 2 - 3 days) for which the embryos can be grown. If further development is required, it may be possible to use exo utero preparations (Chapter 2) or to perform i jections in 1ttero (Chapter 3). Both of these alternative techniques permit development to term and beyond (following Caesarean delivery), though the range of manipulation they permit is more restricted than with embryos in culture. A review of the range of culture techniques that are available and the results that may be expected from them are described by New in the Introduction to this volume.
Introduction of cloned DNA into plant pathogenic bacteria
Authors: Keen Noel T , Shen, Hao , Cooksey Donald A
Published: November 2023

The introduction of cloned DNA into an organism of interest is an essential part of modern biological experimentation. This may be a vexing problem with poorly characterized organisms containing various uncharacterized plasmids, restriction/modification mechanisms, and recombinational systems. Not surprisingly then, bacterial plant pathogens have often posed difficulties in the introduction of exogenous plasmid DNA. While classical chemical transformation methods have been reported for some of these organisms (see, for example, refs 1 and 2), several pathogenic bacteria have not been successfully transformed or, if so, the frequencies are exceedingly low. Transducing phages are not generally useful with plant pathogens for the introduction of DNA. One clever approach, however, has been the introduction of the cloned E.coli lamB gene into other Gram-negative bacteria (3). These cells expressing LamB protein are then receptive to transduction with packaged phage lambda cosmid clones. Since phage packaging is required, this is not a general approach for the introduction of DNA into plant pathogenic bacteria, but may be of use for the establishment of cosmid libraries.
Physiological approaches to the study of glutamate receptors
Authors: Jahr C E , Westbrook G L
Published: November 2023

Our understanding of glutamate receptors and their role in central excitatory neuro transmission has dramatically increased in the past five years. This has been driven by rapid technical improvements in the pharmacological agents available for studying glutamate receptors, as well as by the refinement of in vitro preparations. The purpose of this chapter is to survey some of the methods that have been useful in studies of glutamate receptors and ion channels in the past few years. Obviously, many of these techniques are common to the study of any receptor or ion channel, especially in the central nervous system (CNS). Thus no attempt will be made here to comprehensively review all the techniques that may have application to glutamate receptor studies. Rather we will focus on techniques which have influenced major issues in excitatory amino acid physiology and pharmacology.
Identifying and characterizing stretch-activated ion channels
Authors: Erxleben Christian , Ubl Joachim , Kolb Hans-Albert
Published: November 2023

The first observation of stretch-activated (SA) channels was probably accidental. It is a common practice to use moderate suction on the tubing connected to the patch-pipette to facilitate the formation of a gigaohm seal. The first reports of SA-channels were those of Guharay and Sachs (1984) (1) on chick myotubes and of Brehm et al. (1984) (2) on embryonic Xenopus muscle. Subsequently, SA-channels have been found in a variety of biological systems, ranging from bacteria, yeast, and plant cells, over molluscan heart muscle to vertebrate oocytes, lens epithelia, endothelial cell lines and kidney cells (for reviews see: refs 3, 4 and 39). The most recent reports also include K+-selective SA-channels in growth cones of neurons, where they co-exist with stretch-inactivated channels (5), in human fibroblasts (6), embryos of fresh-water fish (7) and in stretch-receptor neurons of the crayfish (8, 9).
In situ hybridization with synthetic DNA probes
Authors: Wisden William , Morris Brian J , Hunt Stephen P
Published: November 2023

In situ hybridization is the colloquial term for a now standard technique used to detect mRNA (e.g. coding for a peptide transmitter or neurotransmitter receptor) in the neuronal or glial cells where it is being synthesized, using a labelled nucleic acid probe of complementary sequence to a portion of the mRNA.
Optimization strategies in microbial process development and operation
Authors: Bull Alan T , Huck Teresa A , Bushell Michael E
Published: November 2023

The predominant theme of the Society’s 29th Symposium a decade ago was the supplanting of empirical discovery by rational design in the field of microbial technology. Thus, the possibilities were seen for predicting whether or not a given material, or reaction, could be produced microbiologically; what type of organism would be most suitable for the process, and how it might be sought and discovered; the conditions under which the organism should be grown in order that it express the desired property; and how a particular property could be modulated by choice of environmental conditions or nutrient status, or by genetic manipulation of the organism (Bull et al., 1979). Our aim in this paper is to encourage further the use of techniques and thinking that provide the best chances of achieving these objectives.
Principles of Protein Stability. Part 2—Enhanced Folding and Stabilization of Proteins by Suppression of Aggregation in Vitro and in Vivo
Authors: Wetzel Ronald
Published: November 2023

1. Introduction Protein folding is most conveniently studied under conditions where unfolding is reversible, and much of what we know about the forces that control the formation of native protein structure is derived from studies on reversible systems. But outside of the biophysical laboratory—and, for that matter, often within it—proteins are more mortal. Irreversible side-reactions limit the stabilities and folding efficiencies of many proteins under native conditions in vitro. Similar side-reactions also play roles in protein folding in vivo. A major class of irreversible side-reactions, which can occur both in vivo and in vitro, is chemical modification. Proteolysis is of major importance in vivo, and can also occur during in vitro folding of proteases. Other chemical reactions occur as well, especially in the in vitro inactivation of proteins (1). These pathways will not be discussed in this chapter. The topic under consideration here is aggregation, which in many ways is more subtle than other non-productive pathways. Aggregation can take place under more mild conditions, and generally leaves the protein intact chemically. Aggregation and precipitation are often thought of as the consequences of failed folding experiments, whereas they are often the cause of failure. An appreciation of the roles of aggregation and precipitation is a critical prerequisite for effectively designing and monitoring the progress of formal folding and stability experiments in the biophysics lab, as well as for the everyday production and handling of proteins in the laboratory and the factory. This problem thus touches protein engineering both in the mundane aspect of obtaining well-behaved protein variants from heterologous expression systems, and in the use of such variants in folding and stability studies.
Expression of Proteins in Prokaryotic Systems—Principles and Case Studies
Authors: Yarranton Geoffrey T Y , Mountain Andrew
Published: November 2023

1. Introduction The rapid developments in recombinant DNA techniques have resulted in the identification and isolation of many novel genes, some of known function and some of unknown function. Almost invariably there is a need to express the gene in a heterologous cell system in order to produce:material for structure-function studies diagnostic reagents, e.g. monoclonal or polyclonal antibodies material for in vivo activity testing Several alternative systems for the expression of foreign genes have been developed including mammalian cells, insect cells, fungal cells, bacterial cells, and transgenic animals or plants. The choice of expression system for a given gene depends upon the likely properties of the encoded protein, e.g. protein modifications needed for biological activity, as well as the objective of the study, e.g. structure-function analysis or production of diagnostic reagents. Other important considerations for the investigator are the facilities available, times and cost involved in generating the amounts of recombinant protein required. The most widely used and convenient system for the production of foreign proteins remains that based on the simple prokaryote, Escherichia coli.
Biomonitoring: an epidemiologic perspective
Authors: Cuzick Jack
Published: November 2023

Analytical epidemiologic studies are of two main types: case-control studies and cohort studies. New methods for assessing exposure and genetic damage from environmental carcinogens by protein and DNA adducts have something to offer each type of study, but because these study designs work at the two poles of natural history of the disease, the contributions are quite different.
Extraction of nucleic acid from fresh and archival material
Authors: Jackson David P , Hayden Jeremy D , Quirke Phil
Published: November 2023

The molecular biological study of fresh clinical material is relatively straight-forward, since good quality DNA or RNA can be easily extracted. However, the nucleic acid from archival material tends to be impure, of low molecular weight, containing single-stranded nicks, and hence is not as amenable to analysis. The sensitivity of the polymerase chain reaction (PCR) allows small quantities of poor quality genetic material to be tolerated. As long as there is sufficient single-stranded DNA or RNA to bridge the distance between the 5’ ends of the two oligonucleotide primers, then amplification can take place (1). Hence, the PCR could be used for the retrospective molecular biological study of a wide variety of pathological archival material. This also has implications for the extraction procedures that may be used to provide template DNA or RNA for the PCR, since very simple, rapid techniques should provide suitable quality template material.
Protein targeting to mitochondria
Authors: Wienhues Ulla , Koll Hans , Becker Karin , Guiard Bernard , Hartl Franz-Ulrich
Published: November 2023

Over the last decade mitochondrial protein import has developed into one of the principal experimental systems for studying the general principles of intracellular protein sorting and the mechanisms involved in the translocation of proteins across membranes (for review, see references 1-5). Most mitochondrial proteins are synthesized as precursors on cytosolic polysomes and are subsequently imported into the pre-existing mitochondria which propagate by growth and division (Figure 1). The cytosolic precursors of mitochondrial proteins are maintained in a loosely folded, translocation competent conformation by the interaction with chaperone components such as cytosolic hsp70. In general, precursors carry amino-terminal pre-sequences which contain necessary and sufficient information for the targeting to mitochondria. Mitochondrial targeting signals are typically positively charged and rich in hydroxylated amino acids. Precursors interact with receptor proteins of 19 and 72 kd (in Neurospora crassa) on the surface of mitochondria. For the insertion into the outer membrane most precursors appear to use a ‘general insertion protein ‘ (GIP) in the outer membrane. Proteins of 38 kd in N. crassa and 42 kd in Saccharomyces cerevisiae have recently been identified which may constitute at least part of GIP.
Techniques in nuclear protein transport
Authors: Dingwall Colin
Published: November 2023

The transport of macromolecules across the nuclear envelope occupies a central place in the physiology of the cell. Interest in the study of this process has been relatively slow in developing but it now represents a major area of investigation. Recently there have been a number of demonstrations of regulated nuclear entry of proteins involved in cell cycle control, cellular transformation, and determination of cell fate in development. A variety of biochemical events are thought to bring about this regulated entry. These include protein phosphorylation or proteolysis to release a nuclear protein from an attachment site in the cytoplasm or reveal a cryptic nuclear targeting sequence. Unravelling the molecular basis of this control will require reliable techniques and many of those in current use are described in this chapter.
Genetic engineering of animal cells
Authors: Macdonald Caroline
Published: November 2023

The commercial exploitation of mammalian cells has largely been for the production either of viruses for vaccines, or for monoclonal antibodies. This situation is changing, however, as the techniques for introducing and expressing foreign genes in animal cells become more routine. This chapter will review the most commonly used methods for introducing genes into animal cells and compare their advantages and disadvantages. It will also describe the strategies for constructing expression vectors and for selecting cell lines expressing heterologous genes at a high level.
Types I, III, and V collagen and total collagen from cultured cells and tissues
Authors: Peterkofsky Beyerl Y , Haralson Michael A , Dimari, Samuel J , Miller Edward J
Published: November 2023

The fibre-forming collagens—types I, III, and V—constitute the majority of collagens in non-cartilagenous tissues. The common structure of these molecules is typified by the type I molecule, which is illustrated in Figure I (1). The general and specific features of these collagen types have been the subject of many reviews (2—10) and will not be exhaustively discussed in this chapter. However, several unique structural aspects of these molecules that are exploited for their isolation, identification, and quantification are briefly reviewed for purposes of orientation. First, the molecules possess long, non-interrupted triple-helical regions. This feature confers upon the molecules resistance to proteolytic digestion, particularly to the enzyme pepsin. Second, these molecules, as do the other collagen types, contain multiple repeats of the sequence —R-Pro—X—Gly—Pro—R—.
Patch clamp methods for single channel and whole cell recording
Authors: Standen N B , Stanfield P R
Published: November 2023

The patch clamp technique was first described by Neher and Sakmann (1) and further developed by Hamill et al. (2). It has led to major and widespread advances in the electrophysiological analysis of the behaviour of cell membranes for two main reasons. First, it has allowed the resolution of currents through single ion channel proteins in native cell membranes. This has led to detailed studies of their permeability and kinetic properties, pharmacology, and modulation, as well as providing powerful new criteria for the classification of channels, leading to the identification of several new types. Secondly, patch clamp has increased the range of cells suitable for electrophysiology to include many (especially mammalian) types too small or fragile for previous techniques.
Nucleic acid and protein sequence management
Authors: Hodgman T C
Published: November 2023

Computers have been used to store, manipulate, and analyse biological sequences since the mid 1970s. Indeed, a major industry has grown up to provide this area with a wide variety of specialist software and hardware. Several textbooks have already been written on different aspects of sequence management (1-7), so this chapter offers guidelines for the novice on the types of software available and how their results should be interpreted. It would be impractical to catalogue all the programs available on the different machines, because the list is very large and subject to revision every month. Hence, this text emphasizes the more well known and widely used packages and programs. Microcomputer design and capability are advancing rapidly. While the standard machines of five years ago-in particular the IBM PC and Apple II-are still capable of general sequence management, they are now considered somewhat slow or restrictive, especially with regard to graphics. Their descendantsthe IBM PS/2 and Apple Macintosh-do a much better job, though while it is almost trivial to transfer software on to the PS/2 the same cannot be said of the Macintosh. The Macll, however, has the option of using the Unix operating system, for which a wide variety of software is available. At the other end of the range, technological development has enabled minicomputer systems to be miniaturized to a microcomputer scale.
Microcomputers in the comparative analysis of one-dimensional electrophoretic patterns
Authors: Costas Menelaos
Published: November 2023

An extensive range of electrophoretic techniques is available to the biomedical scientist wishing to separate complex mixtures of macromolecules. The majority of these techniques are variants on a single theme and are generally dependent on the principle that charged macromolecules move under the influence of an electric field. The rate of migration for each molecule differs, and is dependent on the charge density of the molecule concerned. Separation is usually achieved in a solution stabilized within a solid supporting matrix in order to fix the individual macromolecules to their final position without consequential loss in resolution due to post-electrophoretic diffusion effects. After electrophoretic separation, the macromolecules are visualized by staining to produce, in essence, a pattern consisting of a series of bands in differing positions (mobility) and with different densities.
Chemical derivatization in gas chromatography
Authors: Watson David G
Published: November 2023

Gas chromatography (GC) of volatile or non-polar compounds may be carried out without derivatizing the sample, indeed derivatives of compounds such as hydrocarbons or halogenated hydrocarbons cannot easily be prepared. It is possible to analyse polar compounds such as carboxylic acids and amines, without prior derivatization, on polar GC phases such as those based on polyethylene glycol. However, derivatization is useful in many instances where it may: For these reasons, GC with derivatization is less frequently employed in quality control applications, where the purity of a single substance or the components in a formulation are being determined, than for instance high pressure liquid chromatography where derivatization is usually not necessary.
Biosynthesis of glycosyl phosphatidylinositol membrane protein anchors
Authors: Field Mark C , Menon Anant K
Published: November 2023

Different hydrophobic interactions are responsible for anchoring proteins to cell membranes. Classically, stretches of apolar amino acids in the protein itself are sufficiently hydrophobic to guarantee membrane association. In other cases membrane association is facilitated by fatty acids or complex lipids covalently linked to the protein. This chapter is concerned with the biosynthesis of a class of complex inositol-containing glycophospholipids (G-Pis) that serve as membrane anchors for a variety of eukaryotic cell surface proteins, including cell adhesion molecules, enzymes, complement regulatory proteins, lymphoid antigens, and protozoal coat proteins.
Modern Machine-Aided Methods Of Oligodeoxyribonucleotide Synthesis
Authors: Brown Tom , Brown Dorcas J S
Published: November 2023

During the 1980s the development of efficient automated methods of solid phase oligonucleotide synthesis has had a major impact in the fields of molecular biology, biotechnology, and biological chemistry (1). Advances in nucleic acid chemistry have made possible the synthesis of oligonucleotides with modified backbones, non-standard bases, or with non-radioactive labels attached to the 3’-or 5’-termini (2). The synthesis of ribo-oligonucleotides has recently become possible and this will undoubtedly have far reaching implications in molecular biology (3, 4). New techniques of DNA sequencing have now been fully automated and the polymerase chain reaction (PCR) has facilitated the detection and amplification of very small amounts of natural DNA (5). Novel and imaginative applications and modifications of oligonucleotides emerge at a remarkable rate and this is clearly a very exciting era for nucleic acid chemists. Some of the most significant recent advances in the field of synthetic oligonucleotides are referred to in subsequent chapters.
Rapid detection of cyanogenic glycoside in malted barley
Authors: Cook R , Oliver W B
Published: November 2023

The occurence of trace amounts of ethyl carbamate in some distilled spirits may be controlled through the use of barley cultivars which lack the ability to produce cyanogenic glycoside during malting. A rapid method is described which enables barley to be screened for this property. The successful development of this simple technique is shown to depend upon the properties of the glycoside and its distribution within the kernel. The sensitivity of the method enables individual grains to be assessed and, examples are given to illustrate applications of the method in barley breeding and varietal identification.
Formation and fate of sulphur volatiles in brewing
Authors: Walker M D
Published: November 2023

A method, based on capillary gas chromatography, has been developed for the quantitative measurement of low levels of flavour-active sulphur volatiles in fermenting wort and beer.
Evaluation of dehusked malt and its influence on the brewing process and beer quality
Authors: Isoe Akira , Kanagawa Kazuo , Ono Miyoko , Nakatani Kazuo , Nishigaki Mamoru
Published: November 2023

According to the dehusked degree, the content of polyphenols in congress wort could be decreased by 10 to 50% The beer, which was brewed with 25% and 50% of the dehusked malt, showed smoother taste and better stability than the beer brewed with normal malt. Such dehusked malt was applied for the development of a new product.
Rheological studies on wheat arabinoxylan: effects of structure and molecular size
Authors: Izydorczyk Marta S , Biliaderis Costas G
Published: November 2023

A water-soluble wheat arabinoxylan was fractionated by stepwise ammonium sulphate precipitation. Compositional analyses, 13C-NMR, gel filtration chromatography, and viscosity measurements revealed that the fractions differed in molecular structure and size. Oxidative gelation (H202/peroxidase) of the arabinoxylan fractions was followed by dynamic rheometry. The gelling potential of arabinoxylan was directly related to the molecular size and the content of feruloyl residues; a correlation between rigidity of cross-linked arabinoxylan gels (as manifested by G’) and intrinsic viscosity (17) of the arabinoxylan fractions was found. The gelling ability was also more pronounced with fractions having a low ratio of ArafXyl and a smaller amount of doubly substituted xylose residues on the chain backbone.
Manipulation and control of mixed polysaccharide/ milk protein gels
Authors: Hart Robert J , Lynch Gerard , M.Dea Iain C , Morris Edwin R
Published: November 2023

Interactions of casein fractions with carrageenan’s have been studied using circular dichroism, polarimetry and small-strain oscillatory rheometry. Circular dichroism indicates the ability of K-carrageenan to alter the conformation of K-casein (but not of a or B-variants) even in the absence of calcium ions. Addition of K-casein to K-carrageenan causes a dramatic increase in the storage modulus (G’), indicating the formation of gel structure. The melting and setting temperature of this gel can be varied in the range 25°-S0°C by suitable control of calcium ion content. Parallel studies of optical rotation suggest that the melting behaviour of the mixed gel is related to the helix-coil transition of the carrageenan.
DSC study on the gel-sol transition of mixed gels of agarose-gelatin, kappa-carrageenan and funoran-gelatin
Authors: Nishinari Katsuyoshi , Kohyama Mineo WataseI Kaoru , Moritaka Hatsue
Published: November 2023

The gel-sol transition of mixed gels of agarose-gelatin, kappa-carrageenan-gelatin and funoran-gelatin was studied by differential scanning calorimetry(DSC). Endothermic peaks accompanying the gel-sol transition in heating DSC curves were analyzed by a zipper model approach. The results suggested that kappa-carrageenan and gelatin, and furan and gelatin promote gel formation of each other whilst agarose and gelatin inhibit each other.
Stability of BSA stabilised oil-in-water emulsions in relation to interfacial rheology. Influence of pH and ionic strength
Authors: Bihan Catherine Le , Lefebvre Jacques
Published: November 2023

The rheological properties in shear of Bovine Serum Albumin (BSA) films adsorbed atadodecane/water interface are studied in shear at three pH’s (4, 4.8, 8) in the absence or presence of NaCl, in parallel with centrifugation-induced coalescence of BSA stabilised emulsions in similar conditions. Rheological characterisation includes the analysis of flow curves obtained in the shear rate range 2.3xl0-3-10 s-1, and of stress relaxation after shearing at 2.3xl0-3 s-1
Bridging flocculation in emulsions stabilised by gelatin and a low molecular weight surfactant
Authors: dickinson Eric , goller michael i. , Wedlock david j.
Published: November 2023

Oil-in-water emulsions (20 vol% mineral oil) have been prepared under standard homogenization conditions at pH 7 using a binary emulsifier system, gelatin+ Tween 20. Laser diffraction particle sizing and light microscopy have shown that the state of droplet aggregation is sensitively dependent on the quantity and composition of the emulsifiers. The behaviour is interpreted in terms of bridging flocculation caused by partial displacement of gelatin from the droplet surface by the more surface-active Tween 20.