bluuebunny/crossref_metadata_2025 huggingface dataset:
A Night to Remember
Authors: Wright Patrick
Published: November 2023

Hackney’s press office appeared surprised that any journalist except the obligatory reporter from the Hackney Gazette should want to attend Wednesday’s council meeting. As things turned out, there were reasons enough. It was to be at this meeting that Medlin Lewis would cross the floor to join the handful of Conservative councillors forming the main opposition in this Labour-controlled London borough. Ms Lewis had been with the council for five years, serving variously as mayor, deputy mayor and chair of the Labour group.
Siraj Izhar’s Public Lavatory
Authors: Wright Patrick
Published: November 2023

‘It’s about black and white—a metaphor.’ Siraj Izhar is trying to help a sceptical BBC television producer who is about to film his latest Strike project for a documentary in which six ‘ordinary’ people from Derry will be seen getting to grips with contemporary art. We are gathered on a pavement in Spitalfields. Cars thunder by on Commercial Street. Homeless men hang around in bunches and an unexpected column of tourists passes by, tracing Jack the Ripper’s bloody trail with the help of a seedy-looking guide. The scene is dominated by the looming front of Christ Church, the vast and largely disused church by Nicholas Hawksmoor that has found such a mixed cult in recent years.
Life annuities
Authors: Pitacco Ermanno , Denuit Michel , Haberman Steven , Olivieri Annamaria
Published: November 2023

Great attention is currently devoted to the management of life annuity portfolios, both from a theoretical and a practical point of view, because of the growing importance of annuity benefits paid by private pension schemes. In particular, the progressive shift from defined benefit to defined contribution pension plans has increased the interest in life annuities, which are the principal delivery mechanism of defined contribution pension plans. Among the risks which affect life insurance and life annuity portfolios, longevity risk deserves a deep and detailed investigation and requires the adoption of proper management solutions. Longevity risk, which arises from the random future trend in mortality at adult and old ages, is a rather novel risk. Careful investigations are required to represent and measure it, and to assess the relevant impact on the financial results of life annuity portfolios and pension plans.
Negotiating Aid
Authors: Whitfield Lindsay , Fraser Alastair
Published: November 2023

The literature on foreign aid consists predominantly of a debate between academics in donor countries and aid practitioners, with many key contributions written or commissioned by donors and focused on identifying ‘best practices’. As a result, aid has been analysed mainly from the viewpoint of donors, focusing on the strategies they should adopt to secure their policy preferences. The sources of leverage for African governments, their perspectives, and the negotiating strategies they chose have been largely neglected. Our main aim is to redress this imbalance. This chapter lays out the framework for conceptualizing aid negotiations between recipient governments and donors.
Oil and Gas in Africa
Authors:   ADB
Published: November 2023

Africa has historically been referred to as a continent with rich natural resources but with high levels of poverty. Indeed, Africa is considered to be well-endowed with minerals, including fossil fuels, although the exact economic potential of these resources is not known. New discoveries of oil and gas in Africa—in locations never before thought to hold such resources in significant amounts—prove that the continent is still “virgin” in many aspects regarding exploration and exploitation of oil and gas resources.
Impact of High Oil Prices on African Economies
Authors:   ADB
Published: November 2023

This chapter reviews evidence of the economic and socio-environmental effects of high and rising oil prices on African countries. In the past, significant increases in the price of oil have led to worldwide economic recessions, such as the 1973 and 1979 energy crises. In many European countries, which have high taxes on fuels, such price shocks could potentially be mitigated by reducing the taxes as fuel costs rise. In many African countries, on the contrary, this option is not feasible, as there is limited taxation on oil products in the first place. In some cases, the oil prices are subsidized. African governments thus face a considerable dilemma: how much of the increase in oil product prices they should “pass through” to consumers. The negative effect of rising oil prices is thus potentially large for net oil-importing countries.
Fermentation
Authors: McNeil B , Harvey L M
Published: November 2023

Fermentation is important in the areas of both academic and industrial research. This book is a collection of methods and techniques covering the setting up and use of fermentation units. The emphasis is on the use of small-scale fermenters and the interdisciplinary nature of fermentation itself. As well as being of widespread interest to academic researchers, this technology has many important industrial applications, as evidenced by the affiliations of the authors (Hoechst, ICI, and Beecham Pharmaceuticals).
RFLP analyses and gene tagging for bacterial identification and taxonomy
Authors: Gabriel Dean W , Feyter Robert De
Published: November 2023

Accurate identification of bacterial isolates to species, subspecies, and pathovar groups coupled with an objective bacterial taxonomy are of paramount importance to microbiologists and plant pathologists. Highly refined genetic technologies facilitate high resolution distinctions such that taxonomies based on phenotype (phenetic taxonomies) are proving to be artificial by genetic standards. The availability of powerful analytical tools such as DNA sequencing, DNA-DNA hybridizations, multilocus isozyme analyses, and restriction fragment length polymorphism (RFLP) hybridization analyses is influencing change towards phylogenetic based taxonomies. Accurate strain identification is also important for epidemiological and ecological monitoring purposes. The simplest and most direct method for the short-term monitoring of known strains released into the field is by direct examination of RFLPs of plasmid DNAs. Plasmid instability may, however, prove a problem to long-term monitoring and other methods, such as monoclonal antibody batteries, polymerase chain reaction analyses, specific DNA probe analyses, or the use of genetically marked strains may prove to be superior. When large numbers of samples must be processed the latter two methods may be the best choices. The selection of antibodies, primers, probes, and/or marking strains requires significant preliminary work. This chapter discusses the use and limitations of RFLP technology and genetically marked strains for taxonomy, identification, and epidemiology of plant pathogenic microbes.
Methods of Structural Analysis of Proteins. Part 2—Nuclear Magnetic Resonance
Authors: Clore G Marius , Gronenborn Angela M
Published: November 2023

1. Introduction Over the past few years solution nuclear magnetic resonance (NMR) spectroscopy has been established as the only viable alternative to X-ray crystallography (see Chapter 1) for determining three-dimensional structures of proteins up to 10 kDa (1, 2). The main source of geometric information used to determine structures by NMR lies in short (<5 A) approximate interproton distance restraints obtained from nuclear Overhauser enhancement (NOE) measurements, supplemented by torsion angle restraints derived from three-bond coupling constants. Methodological advances in the past year, particularly with respect to obtaining many more interproton distance restraints from the NOE data, coupled with stereospecific assignments and torsion angle restraints derived from systematic conformational search procedures, has led to a considerable increase in the precision and accuracy of the structures that can be obtained by NMR (3, 4).
Molecular Sequence Data-Bases and Their Uses
Authors: Boguski Mark S , Ostell, James , States David J
Published: November 2023

Introduction There are many aspects of nucleic acid and amino acid sequences pertinent to protein engineering. For example, promoters and other types of non-coding regulatory DNA and RNA can be studied and manipulated with the purpose of designing transcription and translation units for maximal expression of natural or mutated proteins in heterologous systems. However, this topic is covered elsewhere in this volume (Chapters 13–15) and for present purposes we consider nucleic acid sequences only as additional sources of conceptually translated protein sequences. The study of natural sequence variation in protein families also has obvious implications for protein design and engineering but will not be dealt with here (see Chapter 14).
From Protein Sequence to Structure
Authors: Bates Paul A , Sternberg Michael J E
Published: November 2023

1. Introduction Knowledge of the three-dimensional structure of a protein is central to the systematic engineering of the function and stability of the molecule. The structural information can define the residues forming the binding site for a ligand or identify residues to change in order to alter thermal stability by the introduction of disulphide bridges or salt bridges. The most accurate method of obtaining protein structure is X-ray crystallography but, despite improvements in methodology (see Chapter 1), the technique remains dependent on obtaining crystals suitable for analysis. Recently, NMR (see Chapter 2) has been used successfully to determine the structure of proteins of up to 200 residues. However, at the present time, fewer than 100 new protein structures will be revealed in a year by these experimental methods, whereas the sequences of at least 1000 proteins are obtained from their cloned gene sequences. Accordingly, computer algorithms are being used increasingly to obtain structural information for a protein from its sequence. In addition, these algorithms can provide guidelines for modifications to experimentally determined structures and are essential for the design of novel proteins.
Human biomonitoring for somatic chromosome damage
Authors: Sorsa Marja , Salomaa Sisko , Ojajarvi Anneli
Published: November 2023

Cytogenetics for somatic chromosome damage has its special relevance in human exposure assessment not only concerning cancer risk as a potential hazard, but also other health impairment conditions related to genetic effects. Chromosomal changes obviously play an important role in the activation of proto-oncogenes but, also, a substantial part of human genetic disease burden involves chromosomal changes similar to the ones observable in the peripheral blood lymphocytes, the cell type mostly used in surveillance studies. Besides, a common characteristic of clastogenic agents, i.e. compounds inducing structural chromosome aberrations (CA) is that they generally also produce point mutations, base deletions, transversions or transitions.
An assessment of the circulating T-lymphocyte 6-thioguanine resistance system for population monitoring
Authors: Cole Jane , Arlett Colin F , Green Michael H L , Norris Paul G , Price Margaret L
Published: November 2023

Circulating lymphocytes from peripheral blood are the obvious choice of material for human population monitoring to assess DNA damage in somatic cells. For many years, chromosome damage has been assessed by scoring chromosome aberrations (CA) and micronuclei (MN) in mitogenstimulated lymphocytes. Such end-points frequently represent lethal events, and in 1979 Strauss and Albertini developed an autoradiographic technique for detecting heritable mutations at a specific locus, the Xlinked hypoxanthine phosphoribosyl transferase (hprt) gene. Subsequently, this method was modified to avoid obvious artefacts but a major disadvantage remains in that the mutant status of the hprt variants scored cannot be verified.
32P-Postlabelling assays of DNA adducts formed in vitro and in vivo with benzene and its metabolites: new assays to measure adducts as 5’-P-labelled dinucleotides and nucleoside monophosphates
Authors: Reddy M Vijayaraj , Blackburn Gary R , Bleicher William T , Irwin Susan E , Mehlman, Myron A , Mackerer Carl R
Published: November 2023

Studies of Maltoni et al. and the National Toxicology Program have shown that the chronic high-dose adminstration of benzene to rats produces solid tumours in Zymbal gland, oral cavity, nasal cavity, and possibly mammary gland. Also, the chronic administration of benzene to rodents causes aplastic anemia and other blood disorders. Investigations on the mechanism(s) underlying the carcinogenic and toxic actions of benzene have focused on its conversion to toxic metabolites and its ability to damage DNA.
Development of techniques to monitor exposure to vinyl chloride and ethylene oxide
Authors: Santella Regina M , Young Tie Lan , Habraken Yvette , Ludlum David B
Published: November 2023

Ethylene oxide (EO), one of the top 50 chemicals in use in the US, is a fumigant and sterilant in hospital equipment as well as an intermediate in the synthesis of a number of industrial chemicals.
The relationship between alkylation of haemoglobin and DNA in Fischer 344 rats exposed to vinyl chloride
Authors: Watson W P , Potter D , Blair D , Wright A S
Published: November 2023

The measurement of the amounts of products formed between genotoxic chemicals, or their metabolites, and macromolecules in body fluids has been proposed by Ehrenberg and colleagues as a basis for determining individual exposures to such agents and improving the quality of risk assessment (for a review see ref. 1).
Protein expression and processing
Authors: Dickson Alan J
Published: November 2023

Understanding the mechanisms which control the generation of proteins from cells in culture is of interest for both commercial and intellectual reasons. For example, the availability of monoclonal antibodies from hybridoma cell lines has great clinical and diagnostic implications. Recent advances in the technology involved in gene manipulation have made it possible to utilize mammalian cells for expression of proteins of defined nature. A number of proteins of possible therapeutic value have been generated from genetically engineered mammalian cells. The procedures for the production of recombinant proteins are designed to ensure the absence of contaminants often found in protein preparations purified from human or mammalian sources by conventional procedures. Several pharmaceutical companies have invested a great deal of time and effort in large-scale production of specific recombinant proteins, including tissue plasminogen activator, blood clotting factors (factors VIII and IX), and erythropoietin (see Chapter 10).
Centrifuges and rotors
Authors: Roth Hans , Rickwood David
Published: November 2023

Centrifuges can be classified on the basis of a number of different criteria. However, centrifuges are usually classified on the basis of their speed since this is a fairly good indicator of the centrifugal force (RCF) which can be generated by the machine and hence also indicates the range of possible applications that the centrifuge can be used for. Table 1 gives a summary of some of the principal classes of centrifuge, namely low-speed, high-speed, and ultracentrifuge. While these classes used to be quite distinct, over the years the boundaries have become somewhat more blurred in terms of the size, costs, and applications of the different machines. The primary interest of the user is simply an answer to the question ‘what can I use it for?’. The answer to this question is more complicated than it might seem. As an example, plasmid DNA can be prepared on isopycnic CsCl-ethidium bromide gradients (see Section 3.3.1 of Chapter 4) using one of the high-performance high-speed machines generating 100 000g. However, using such machines gradients need to be centrifuged for at least 30 h while using an ultracentrifuge the same isolation could be done in 2-5 h depending on the type of rotor used. Hence the time available to the researcher can also be a crucial factor in choosing the type of centrifuge for a particular application.
The concept of cleaner production as applied to industry
Authors: Huisingh (P) Dr Donald
Published: November 2023

Examples drawn from industrial case studies of “Cleaner Production”, are presented to illustrate the advantages of these approaches over the pollution control approaches that are utilized most extensively today. Recommendations pertaining to the brewing and food industries are made on how these environmental challenges can be used to guide the development and implementation of corporate policies and procedures that are economically and environmentally better.
Arabinoxylans in barley, malt and wort
Authors: Victor R J , Angelino S A G F , Voragen A G J
Published: November 2023

Arabinoxylans from barley endosperm cell walls may impair the wort preparation process. They were isolated from dehusked barley and malt, and from sweet wort. Their glycosidic linkage composition was determined, showing the presence of a large amount of C2-substituted xylopyranose. On enzymic degradation of these arabinoxylans, a range of oligomers with various degrees of polymerization below 7 was formed, which accounted for half of the starting material. The residue consisted of arabinoxylans with an increased arabinose content. Differences between the arabinoxylan fractions studied were negligible with respect to sugar and linkage composition as well as enzymic degradability.
Improvements in micronised wheat production
Authors: South J B
Published: November 2023

The use of significant quantities of wheat as a brewing adjunct has been limited by mash filtration problems particularly in rapid brewing schedules. Studies using micronised wheat suggest that this may be due to denaturation of proteins during micronising, with the formation of large disulphide- linked aggregates, which then undergo rearrangement during mashing and impede run-off. Conditions for minimising protein denaturation have been established leading to improved performance both in laboratory scale and brewhouse mashes.
Prediction of yeast fermentation performance using the acidification power test
Authors: Mathieu Ch. , Berg L van den , Iserentant D
Published: November 2023

The Acidification Power (AP) Test allows, by monitoring the acidification of the medium, to estimate the physiological condition of yeast. The AP of the yeast was measured during industrial fermentation. The AP shows a sharp decline at the beginning of the fermentation, compared to the AP of the pitching yeast.
Yeast dehydrogenase activities in relation to carbonyl compounds removal from wort and beer
Authors: Collin S , Montesinos M , Meersman E , Swinkels W , Dufour J -P
Published: November 2023

Carbonyl compounds of wort and beer are well-known off-flavours. Their removal by yeast contributes to improved beer flavour and to less susceptibility to staling. Carbonyl marker compounds have been followed simultaneously with deshydrogenase activities of yeast: acetoin/diacetyl (diacetyl pathway), hexanal/nonenal (lipid route), 3-methyl-butanal (Strecker pathway) and acetaldehyde (glycolysis). Potential use of deshydrogenase activities measurements has been investigated with regard to yeast performances for the removal of carbonyls during fermentation and for the production of non-alcoholic beer using an immobilized yeast bioreactor.
‘Head hunting’
Authors: Haugsted Carsten , Erdal Kenneth
Published: November 2023

Digital video image analysis is used in a foam quality assay which provides a consumer related, yet objective impression of the overall beer foam behaviour. The results introduce a somewhat expanded and varied foam quality concept, and thereby a reasonably sound basis for decisionmaking regarding the means by which to optimize the foam quality of the product. “Field tests” illustrate the influence of the raw materials, the production processes, and the addition of pre-isomerized hop products as questioned by the brewer.
Foam stabilizers may contribute to beer haze
Authors: Outtrup Henrik
Published: November 2023

Use of propylene glycol alginate (PGA) for foam enhancing may concomittantly imply haze problems due to decomposition of PGA molecules during beer storage. From model studies of the thermodynamics of this process it is suggested that the haze potential is predominantly introduced by free carboxylic groups which appear after deestrification of PGA molecules.
Simultaneous in-line measurement of dissolved carbon dioxide, nitrogen and oxygen using vapour pressure and mass spectrometry
Authors: Forrest LS , Cuthbertson R C , Dickson J E
Published: November 2023

Continuous and simultaneous in-line measurement of dissolved carbon dioxide, nitrogen and oxygen is desirable as each can affect the taste and appearance of beer. A system which combines the measurement of vapour pressure and molecular ratio has yielded accurate and reliable estimates of dissolved gas concentrations in plant trials. The necessary measurements were obtained by combining the phase separation properties of silicone rubber membranes with an industrial Quadropole Mass Spectrometer. The instrument can be fitted directly in the beer stream, is robust and hygienic, and thus can form the basis of a system to control the injection of carbon dioxide or nitrogen into beer.
Das Verhalten toxischer Schwermetalle im Brauprozess
Authors: Leubolt R , Urban A , Huber C , Püspok J
Published: November 2023

Heavy metals, which come into brewing process from the raw materials and from the materials of construction of the brewing plant, are precipitated with spent grains, break and yeast. Consequently neither the mashing (infusion, decoction) nor differences in raw materials (petal hops, hop pellets, hop extract) have any noticeable effect on the concentration of metals in the finished beer. Heavy metals are precipitated to a large degree as semi metals (arsenic, antimony, bismuth). The greatest amounts of heavy metals are recovered from spent grains. The elements taken up by yeast during fermentation do not have any noteworthy effect on its activity when it is reused.
Improvements in twist-off crowns applied to refillable beer bottles
Authors: Gracey D E F , Miller S , Rutledge T , Brown S , Lumsden J , Campbell G D , Bouchard G J
Published: November 2023

In 1984 a significant packaging initiative for the brewing industry in Canada involved twist-off crowns applied to reusable bottles. While gaining immediate acceptance by the consumer, these were not entirely satisfactory with respect to packaging quality objectives. Subsequent trials resulted in the adoption of a crown design which improved the handling qualities for the customer although other sporadic quality concerns remained.
Molecular tnechanistns of antigen processing
Authors: Monaco John J
Published: November 2023

The mammalian immune system has developed methods for the surveillance of both extracellular and intracellular compartments for 'non-self'. This surveillance is effected by antigen-specific receptors on T lymphocytes, whose natural ligands are short peptides complexed with major histocompatibility complex (MHC) encoded class I or class II molecules. MHC molecules are integral membrane proteins, specifically designed-to bind a diverse set of peptides and to deliver them to the cell surface for recognition by T cells. The structural features of MHC molecules and their peptide binding specificities are discussed in Chapter 4 and in recent reviews (1-4). Here I will focus on the molecular mechanisms used to generate MHC-binding peptides and to deliver them to the appropriate MHC molecules.

Early experiments based on the ability to differentially induce or inhibit antigen presentation by MHC class I versus MHC class II molecules suggested that differ ent mechanisms were utilized for the loading of these two classes of molecules with peptides (reviewed in refs 5-7). More recently, it has become possible to isolate and characterize the peptides found naturally associated with MHC molecules; the data derived from such studies are generally consistent with the notion of separate pathways for the derivation of class I- and class II-associated peptides. In general, MHC class I-associated peptides are derived from proteins that are either synthesized on the host cell's own ribosomes (3, 4), or which are artificially introduced into the cytoplasm of living cells (8, 9). In contrast, peptides found naturally associated with MHC class II molecules are derived predominantly from the extracellular portions of cell surface proteins, or from proteins endocytosed from the extracellular medium (10-14).
The complement systent
Authors: Reid Kenneth B M
Published: November 2023

Complement can be activated by two main routes, the classical and alternative pathways (Fig. 1). The major complement component C3 (present at 1.3 mg/ml plasma) is common to both pathways. C3 plus the other 12 glycoproteins shown in Fig. 1 and Table 1 constitute the 13 components of the pathways. A recently described third route, the lectin pathway, bypasses the Cl stage of the classical pathway but utilizes all the other components of the classical pathway (Fig. 1). Control of the activated components is mediated partly via at least eight control proteins present in plasma (Table 1) and partly by a variety of membrane-bound control proteins and receptors (Table 2). These membrane proteins bind activated components or fragments of the activated components generated by further limited proteolysis. Many of the activation and control steps in the system involve the splitting of only one or two specific peptide bonds, and therefore it is useful to identify, at an early stage, which components and control proteins are enzymes.
In situ hybridization histochemistry with radioactive and nonradioactive cRNA and DNA probes
Authors: Lewis, Michaele.
Published: November 2023

In situ hybridization histochemistry has been established as a uniquely powerful tool for the study of gene expression at the level of the single neurone. Although this technique has also frequently been used to confirm immunoreactive neurones as the actual site of biosynthesis of a specific peptide or protein, the greatest utility is apparent in studies of altered gene expression in development, pathology, or due to physiological or pharmacological manipulations. The purpose of this chapter is to provide the reader with established protocols for carrying out in situ hybridization histochemistry with either chemically synthesized DNA probes or biosynthesized cRNA probes, using either autoradiographic or enzyme histochemical methods for detecting hybridized neurones.
The Fortmation of Native Disulfide Bonds
Authors: Gilbert H F
Published: November 2023

The disulfide bond complicates an already complex protein folding problem. The mechanisms that direct the formation of three-dimensional native structure from an unfolded protein must now accommodate the chemistry of disulfide bond formation, a formal chemical oxidation. We will begin with a brief discussion of the chemistry of the disulfide bond.
|Retrons in bacteria
Authors: Lim Dongbin , Lima Tania M O , Maas Werner K
Published: November 2023

There are many genetic elements whose life-cycle depends on a reverse transcriptase (RT). These elements are generally called retro-elements and they are found among viruses, transposons, and plasmids. Such genetic elements are widely distributed in eukaryotes. Although there had been sporadic reports on the existence of RT in bacteria, it was not clear until recently that a similar element was present in prokaryotic organisms. A few years ago the production of an RT was clearly shown in two different kinds of bacteria, Escherichia coli and Myxococcus xanthus. Since then, it has been shown that there are RTs in several genera of bacteria.
Pulsed field gel electrophoresis
Authors: Johnston John R
Published: November 2023

Pulsed field (gradient) gel electrophoresis (here abbreviated to PFGE) (1, 2) allows separation of large DNA molecules, typically ranging in size from 50 to 10 000 kb (10 Mb). Pulsed electric fields in different directions are applied to agarose gels. Since the DNA molecules of the 16 (haploid) chromosomes of Saccharomyces cerevisiae range in size from approximately 200 kb to 3 Mb, they are eminently suitable for separation by this technique. This provides definition of ‘electrophoretic karyotypes’ (3) of strains by sizing chromosomes according to how far they migrate in the gel, after staining of DNA bands with ethidium bromide (EthBr).
Ty insertional mutagenesis
Authors: Strathern J N , Mastrangelo M , Rinckel, L A , Garfinkel D J
Published: November 2023

The use of transposable elements as insertional mutagens is an extremely powerful technology for genetic analysis. This approach was brought to a fine art in prokaryotic genetics and is proving equally applicable in fruit flies, mice, and higher plants.
Use of blotted proteins as immunogens
Authors: Szewczyk Boguslaw , Harper David R
Published: November 2023

In the past a considerable amount of effort has been directed at recovering proteins resolved by sodium dodecyl sulphate polyacrylamide gel electro phoresis (SDS-PAGE) and using them as immunogens. Typically, proteins have been recovered from polyacrylamide gels by either diffusion or electroelution procedures. However, in spite of the numerous variations in these procedures, none of the available methods is entirely satisfactory. Some of the methods require complicated manipulations or specialized equipment, while others lead to poor recovery. To overcome the limitations of elution of proteins from polyacrylamide gels, Parekh et al.
Simulation of the degradation of starch and -glucans during mashing
Authors: Koljonen Tatu , Hämäläinen Jari J , Sjöholm Katharinia , Kettunen Anu , Pietilä Kirsti
Published: November 2023

A simulation program for predicting the hydrolysis of starch and B-glucans during mashing was developed. Given the malt characteristics the program helps in the planning of an appropriate mashing temperature profile for specific wort properties. The simulation model consists of 15 ordinary differential equations. Malt enzyme activities, the carbohydrate concentrations, malt-water ratio, and the mashing temperature profile are given as inputs to the model. Activities of a- and B-amylase, B-glucanase and the concentrations of fermentable sugars, dextrins, starch, and B-glucans in wort are predicted. The model predictions were in accordance with the measurements from laboratory and industrial scale mashings.
Development of a novel control system for solids removal by centrifuge
Authors: Platt David W
Published: November 2023

Centrifuges have been used in the green beer and from beer pre-filter. now also recover rest beer from past to remove solids from wort, from The new generation of centrifuges can tank bottoms and waste yeast. All centrifuges suffer in efficiency if loading. Work, initially on hot wort centrifugation and subsequently on rest beer recovery, has developed a novel system for controlling the feed to the centrifuge to optimise its throughput without overloading.
Purified embryonic motoneurons
Authors: Henderson Christopher E , Bloch-Gallego Evelyne , Camu William
Published: November 2023

Spinal motoneurons have been the object of much study using in vivo models, but their detailed study in vitro has been hindered by the cellular complexity of the spinal cord and the consequent difficulties in reproducibly obtaining pure motoneurons in culture. Since the 1970s different methods have been developed for their enrichment, identification, and/or purification. These methods are based on three major properties of the motoneuron: its localization in the ventral spinal cord, its relatively large size, and the fact that its axon projects to the limb bud from early stages (for a more complete review, see ref. 1). Each method has its special advantages and drawbacks: in general, those methods that give a high degree of purification have relatively low yields. Depending on the experiments to be performed, one method may be preferred to another.
Brain endothelium
Authors: Male David
Published: November 2023

The endothelium of the CNS, including that of the retina is quite distinct from endothelium in other tissues. It has continuous tight junctions, low numbers of vesicles, and a variety of characteristic surface molecules (1). In addition, it lacks many of the molecules found on large vessel or microvessel endothelia from outside the brain. In vivo the electrical resistance across CNS endothelium is in excess of 2000 Ohms/cm2, although the best in vitro cultures only approach 500 Ohms/cm2 (2). The factors which control the expression of this distinctive phenotype are not known, but there may be an astrocytic influence via cytokines (3). In vitro, primary cultures of brain endothelium have been obtained from rat, mouse, bovine, chick, and human brain. Some of the primary cultures have been propagated for extended periods (4), however we have found that many of the distinctive features of rat CNS endothelium are lost after a number of passages.
The study of platelet function in other species
Authors: Poole Alastair W
Published: November 2023

Research into platelet function has largely concentrated upon human platelets for readily apparent reasons. There are however special reasons for the study of platelet function in other species. In particular ex vivo pharmacological and toxicological studies are undertaken in experimental animals, in addition to work involving animal models of human platelet-derived disease or work of primary veterinary interest. This chapter will deal mainly with techniques used to obtain platelet preparations from the various domesticated and laboratory species. In addition information is given about species variation in surface receptor populations. This will enable the researcher to better choose the appropriate animal model for their area of research. Useful further reading for information about laboratory animal handling, including bleeding techniques, includes the recent report of a joint working group on refinement in laboratory animal techniques (1), and a newly published handbook of laboratory animal management (2).
Free Radicals
Authors: Punchard Neville A , Kelly Frank J
Published: November 2023

Free radicals are recognized as being of increasing importance in many aspects of normal cell metabolism and pathological processes. Studies in this area depend critically on techniques that allow detection and measurement of free radicals which have lifetimes in the order of microseconds. This book describes the latest methods for quantifying radicals and their effects, providing detailed protocols, hints and tips for success, troubleshooting comments, illustrative data, and key literature citations. Topics covered include: physico-chemical methods of free radical detection, such as ESR and NMR spectroscopy and pulse radiolysis; biochemical methods, such as intrinsic chemiluminescence, multi-functional digital microfluorography, and salicylic acid and phenylalanine probes; measurement of free radical products, including peroxides, oxidized proteins and products of attack on nucleic acids; and measurement of antioxidants such as glutathione and superoxide dismutase. This book is a unique laboratory manual of techniques for the direct and indirect measurement of both free radicals and the antioxidants that protect against them. All researchers who study free radicals will find this volume invaluable.
Simple solid phase assays of avidity
Authors: Goldblatt David
Published: November 2023

In contrast to affinity, avidity or functional affinity (1) is defined as the interaction between a complex antigen and bivalent antibody. Bivalent binding increases the avidity of any given interaction because it represents a summation of individual monovalent affinities. A variety of biological functions may be mediated by antibodies on the basis of their functional affinity/avidity (2). When investigating the interaction between large, complex antigens, such as bacteria or whole viruses, and complementary antibody, binding is likely to be complex hence measures of avidity may be relevant. This chapter will focus on techniques that measure the relative avidity of such antibody-antigen interactions.
Capillary electrophoresis of peptides and proteins
Authors: Hu Bi-Huang , Martin Lenore M
Published: November 2023

The goal of this chapter is to describe a variety of approaches that may be adopted by the biomedical researcher or bioanalytical chemist to achieve successful separations of peptides and proteins. The list of modes commonly used for the analysis and separation of peptides and proteins by capillary electrophoresis (CE) includes: capillary zone electrophoresis (CZE), micellar electrokinetic capillary chromatography (MEKC), capillary isoelectric focusing (cIEF), and capillary gel electrophoresis (CGE). This list is constantly growing and appears unlimited. The application of CE to answer specific questions which commonly occur in peptide and protein research will also be discussed in this chapter.
Introduction to confocal microscopy
Authors: Shaw P J
Published: November 2023

Optical microscopy is inherently limited in resolution by the wavelength of the light used in the technique; in practice this resolution limit is generally about 0.25 µm. However, there are several factors which make optical methods very powerful. One major advantage comes from the combination of microscopy with fluorescent probes. Almost any biological component can be specifically labelled with a fluorescent tag, sometimes called a fluorochrome, and then imaged by virtue of its fluorescent light emission. The available fluorescent probes include dyes for specific components, such as nucleic acids, reporters for particular ions such as calcium, modified DNA and RNA sequences, which are used for fluorescent in situ hybridization, and antibodies raised against almost any biological component imaginable. Fluorescent detection has the advantage of great specificity and sensitivity.
Targeted integrations
Authors: Torres Raul M , Kühn Ralf
Published: November 2023

Apart from generating deletions, are recombinase also mediates the reverse, integrative reaction by recombining loxP-containing, incoming vectors with a transgene or endogenous sequences harboring loxP sites in the genome. Thus, the capacity of the Cre/loxP system for site specific integration could be used as an alternative for homologous recombination to generate secondary modifications in loxP containing loci in cultured cells. Possible applications of loxP-mediated integration could include, for example, the analysis of different promoter regions into the same genomic site for gene expression studies or the replacement of different mutated exons in structure/function studies. For targeted integration, the loxP containing integration vector must be introduced together with a vector for transient Cre expression, ensuring only limited recombinase activity, to avoid the subsequent excision of the integrated, loxP-flanked DNA segment. In plant cells it has been shown that the equilibrium of Cre¬ mediated inversion can be shifted to one side by using a pair of two different mutant lox sites (42). Certainly this technique will be also used in future to stabilize the products of Cre-mediated integration in ES cells by preventing reexcision. A circular vector containing a single loxP site will be integrated completely (Figure 8), whereas from a linear vector with two loxP sites in the same orientation only the loxP-flanked region will be inserted, probably after extrachromosomal resolution of the floxed segment into a circle with a single loxP site (Figure 8).
Freezing ES cell clones
Authors: Torres Raul M , Kühn Ralf
Published: November 2023

One of the biggest advantages to screening your colonies by PCR is that you can immediately focus on a smaller, more manageable number of colonies. This is because potential homologous recombinants are identified essentially the same day as the colonies are picked. In practical terms this abrogates the need to freeze all of your picked colonies (often 300 colonies) which is a large task. Nevertheless, even with PCR screening the time comes when you (may) need to freeze down ES clones to verify the targeting event by Southern analysis. Depending on the number of potential colonies you need to freeze down (and when screening by Southern analysis, all the colonies are potential recombinants) it is often convenient to freeze down these colonies in 48 well plates. There are different methods to accomplish this which depend on whether you freeze down trypsinized cells or whether you trypsinize after thawing. The former method is given here.
Preparation of rodent monoclonal antibodies by in vitro somatic hybridization
Authors: Dean* Christopher , Shepherdt† Philip
Published: November 2023

Like all great discoveries the development of hybridoma technology by Cesar Milstein and Georges Kohler was essentially a simple procedure namely, fuse an antibody-producing B cell with a myeloma cell line lacking the DNA salvage pathway to generate a continuously growing hybridoma producing a single anti body specificity. In this way the genes coding for the specific heavy and light chains are captured from the B cell and expressed by the hybridoma. The ability to generate monoclonal antibodies (mAbs) with precise antigenic specificities from the spleens or lymph nodes of immune rodents has revolutionized our approaches to biomedical science. Not only have these antibodies opened the doors behind which many gene products would have lain hidden but also they have provided us with exciting new approaches for the diagnosis and treatment of disease.
Isolation and purification of monoclonal antibodies from tissue culture supernatant
Authors: Hale Geoff
Published: November 2023

Why do you need to purify an antibody? There are many possible reasons and the answer will be a major factor in determining the best strategy. Depending on the intended application, different contaminants are important and different degrees of purity are necessary (Table 1). Manufacture of antibodies for human therapy imposes many stringent requirements which are beyond the scope of this chapter. Anyone intending to develop a process for ultimate therapeutic application should seek detailed advice at an early stage.
Antibody production in plants
Authors: Drake* Pascal , Stogert Eva , Nicholson Liz , Christout Paul , Ma Julian K C
Published: November 2023

Plant biotechnology is a rapidly expanding area and in the last five to ten years it has become apparent that plant systems may be particularly valuable for the expression and production of recombinant proteins such as pharmaceuticals, vaccines, and in particular, antibodies. A particular attraction of this approach is the potential for producing these kinds of reagents on an agricultural scale, thereby significantly reducing the costs of production. However there are also many other advantages related to the use of plants. In this chapter, we shall provide a brief review of the advances made in plant expression of antibodies and describe the principles of plant genetic engineering as well as the major techniques that are involved.
lmmunogold probes for light and electron microscopy
Authors: Monaghan Paul , Robertsont David
Published: November 2023

The characteristics of monoclonal antibodies make them particularly suitable for electron microscope (EM) immunolabelling studies. At the high magnifica tion of the EM, any non-specific labelling can be a real problem, and some of the best antibodies we have used are monoclonal antibodies. The principles of immunocytochemistry are essentially the same for both light and electron microscopy. A primary antibody is persuaded to bind to its antigen and this is then detected by a suitable marker. For electron microscopy, a number of mark ers have been used including ferritin and horseradish peroxidase/DAB reaction product. However colloidal gold is an almost perfect marker in this context and alternatives are rarely used. Colloidal gold is particulate and electron dense making it readily visible in the EM. In addition, it does not obscure the underly ing cellular structure, making it easy to localize antigens not just to specific organelles, but regions within these structures. Colloidal gold can be formed in defined sizes and this has implications for its use, particularly in multiple labelling experiments.
lmmunochemical detection of BrdUrd labelled nuclei for monitoring cell kinetics
Authors: Wilson George D
Published: November 2023

Cell kinetics is defined as the measurement of time parameters in biological systems. This chapter describes the use of BrdUrd/DNA flow cytometry to study cell kinetics of cells in culture. The technique involves pulse labelling with BrdUrd after which samples are then taken at time intervals thereafter. The cells are stained with a monoclonal antibody against BrdUrd and visualized by a secondary antibody conjugated to a fluorochrome (usually fluoroscein isothio cyanate, FITC) and counterstained with propidium iodide (Pl) to measure the DNA content, and analysed on the flow cytometer. The simultaneous measure ment of BrdUrd incorporation and DNA content confers sensitivity and versa tility in detection of cell cycle perturbations in response to drugs or radiation and tracing the lineage of a cell within the cell cycle. The speed and quantitative power of the flow cytometer, in conjunction with the specificity and sensitivity of monoclonal antibody techniques, provide the basis for the adoption and success of the BrdUrd technique in experimental and clinical investigations. The use of computer-generated windows facilitates the analysis of any population of cells within any phase of the cell cycle and is not restricted to the ‘mitotic window’, as previously was the case with HTdRfautoradiography.
Assay and purification of protein-serine/threonine kinases
Authors: Grahame Hardie D , Haystead Timothy A J , Salt Ian P , Davies Stephen P
Published: November 2023

It is now very clear that activation of protein kinase cascades (ably assisted by appropriate regulation of protein phosphatases) is the major mechanism by which eukaryotic cells respond to changes in the extracellular environ-ment, such as the presence of extracellular signal molecules or adverse envilronmental conditions. It also appears to be the major mechanism through which cellular processes which occur discontinuously (e.g. DNA replication, mitosis) are switched on and off. Although there are exceptions, protein kinases are usually expressed constitutively, and the key event is activation of pre-existing kinase protein, through phosphorylation, dephosphorylation, binding of an allosteric activator (e.g. cyclic AMP, 5’-AMP, Ca2+, diacylglycerol), or binding of a regulatory protein (e.g. a cyclin). It is therefore very important to develop methods to assay protein kinase activity. In some cases (e.g. MAP kinases) the activating phosphorylation produces a mobility shift on SDS-polyacrylamide gel electrophoresis (PAGE), and the activation can be assessed by Western blotting. However, non-activating phosphorylation events can also produce mobility shifts, and this method can be unreliable or misleading. There is, therefore, no real substitute for the development of direct assays of protein kinase activity. This is the first main topic of this chapter (see Section 2).