bluuebunny/crossref_metadata_2025_split huggingface dataset:
Semiconductor field effect devices
Authors: Winquist Fredrik , Danielsson Bengt 
Published: November 2023

Abstract
The research and development of biosensors is at present in a rapidly growing phase due to the demands from a number of different areas such as clinical chemistry, process industry, agriculture and environmental control. Many different types of biosensor have been developed, and the majority of the biosensor concepts are based on the combination of immobilized enzymes with classical sensors such as photometers, amperometric or potentiometric electrodes, gas electrodes or thermistors. The performance of a biosensor is to a large extent dependent on the sensor elements. One class of sensors that has been developed during the last decade is the solid state chemical sensors; these have properties that make them especially valuable for biosensing purposes. Based on semiconductor technology, they are characterized by their ruggedness, ability to be directly integrated with microelectronics and hence the possibility to be mass fabricated at low cost. One type of such sensor is based on field effect devices, made by the MOS (Metal Oxide Semiconductor) technology. They can be designed either as capacitors or field effect transistors.
Thermometric sensors
Authors: Danielsson Bengt , Winquist Fredrik 
Published: November 2023

Abstract
Bioanalytical calorimetry has many attractive features, which were recognized early in studies using conventional calorimeters. Since biological reactions are usually more or less exothermic, calorimetry offers a general detection method which is insensitive to the optical properties of the sample. Enzymic reactions are associated with rather high molar enthalpy changes in the range of20-100 kJ mol-1 (Table 1) and it is often possible to base measurements on only one enzymic step. This is in contrast to other techniques where the detection is based on, for instance, the change in concentration of coloured reactants. In such cases it is usually necessary to couple the primary reaction with one or several (enzymic) reactions in order to obtain measurable changes.
Cellular assays for lymphoid immune deficiency
Authors: Chapel H , Sewell H 
Published: November 2023

Abstract
Cellular assays for immune deficiency include enumeration of the various cell types involved in the immune response as well as demonstration of their functional abilities. Cellular assays for lymphoid immune deficiency currently involve those of T-cells and B-cells, as well as subpopulations of these cells; in the future, assays for antigen processing cells may also be available. Assays for non specific immune deficiency, i.e. those defects involving phagocytic cells and complement are described in Chapters 3 and 4 respectively.
Molecular characterization of viral infections in vivo
Authors: Ian Lipkin W , Oldstone Michael B A 
Published: November 2023

Abstract
Natural and experimental infections of animals continue to be important as models in studying the pathogenesis and management of infectious diseases. Classically, methods for examining animal models have required extraction of individual tissues for biochemical or histological analysis. Examples of these techniques reviewed elsewhere in this volume include nucleic acid and protein electrophoresis, immunohistochemistry and enzyme linked immunoadsorbent assays (ELISA). Over the past few years, however, we have used a new technology which allows for sensitive and specific detection of nucleic acids and proteins in whole animal sections (WAS) fixed to tape or artificial membranes. The power of WAS technology rests in its unique potential for defining and quantitating the distribution of genes and gene products in an anatomical context. Further, because the entire animal is examined, there is no bias a priori against recognizing novel distributions of genes or gene products. Although we have used WAS primarily for studies in viral pathogenesis, problems in developmental biology, neoplasia, and pharmacology may also be suitable for WAS analysis. WAS would also seem ideal for the investigation of tissue-specific distributions of transcripts and proteins in transgenic animal systems. In this chapter we will discuss our methods for WAS in situ hybridization and protein blotting (Figure 1 ). For illustrations of WAS applications the reader is referred to a recent review (1 ).
Whole-cell voltage-clamp techniques applied to the study of synaptic function in hippocampal slices
Authors: Madison Daniel V 
Published: November 2023

Abstract
When the technique of recording from brain slices came into general use for electrophysiological experiments, a great many predictions were made about the utility of this preparation for conducting experiments on the physiology and pharmacology of the central nervous system. These predictions were based on some substantial and obvious benefits offered by this preparation. Indeed, the slice preparation has been used for experiments that are difficult or imposible to conduct on in vivo preparations. However, as is often the case when foretelling the future, the predictions on the usefulness of the slice preparation proved inaccurate. Indeed, the brain slice preparation has fulfilled its promise perhaps more than anyone could have imagined.
Optical techniques and Ca2+ imaging
Authors: Moreton R B 
Published: November 2023

Abstract
Optical techniques covers a wide range of subjects, including the use of stains or fluorescent dyes to visualize specific subcellular components, and the use of sophisticated image-analysis techniques to identify and map structures in two or three dimensions, or to study cell motility. This chapter concentrates on the use of computer-linked video cameras to capture and analyse optical information. Specific attention is paid to using fluorescent probes to detect and map the intracellular distribution of ions and molecules, especially that of calcium which is providing so much exciting new information about the way in which receptors are used to translate extracellular signals into control of cell metabolism and secretion. For reviews of some other optical techniques, see refs 1-3.
X-ray Microanalysis
Authors: Wendt-Gallitelli Maria Fiora , Isenberg Gerrit 
Published: November 2023

Abstract
1. Introduction The heart ventricular myocyte is one of those contractile cells where movements of Ca couple excitation to contraction, rhythmically every second. According to our textbooks, it is the Ca bound to troponin C (TnC) that controls the actin-myosin interaction (attachment of cross-bridges). Quantitation of this activator Ca, therefore, would help in understanding the way of contractile activation. Besides knowledge of the concentration of Ca-TnC as a function of time, one should know how and from where the activator Ca derives, and how and to where it is redistributed. During the last two decades, a variety of different methods has addressed the above question. The outcome can be summarized in the simplified model of Figure 1. For mammalian ventricular myocytes, the model postulates most of the activator Ca to derive from an intracellular ‘release compartment’ (R) where it has been stored during diastole. Ca-release is induced by Ca-entry during excitation. Ca diffuses from the release compartment into the myofibrillar space along a gradient. Ca diffusion occurs with Ca binding to TnC. TnC competes with sites on the SR Ca-pump which have an even higher Ca affinity. Thus, after a while, Ca has sequestered out of the myofibrillar space into the ‘uptake compartment’ (U). Ca-influx through the sarcolemma is thought not only to trigger SR Ca-release but also to fill the SR with Ca. Vice versa, not all of the released Ca recirculates to the SR, a fraction of 20% might be extruded into the extracellular space. At steady-state beating, the sum of Ca-influx and Ca-efflux should equal.
Ca2+ Buffering: Computer Programs and Simulations
Authors: Fabiato Alexandre 
Published: November 2023

Abstract
1. Introduction The free calcium concentration ([free Ca24]) inside most animal cells is in the nanomolar (nM) to micromolar (µM) range. When one wants to make up a solution simulating the intracellular milieu, it is not possible to obtain such a low [free Ca24) by simply adding a small amount of calcium to the solution.* In fact, the total calcium contamination of the other salts contained in the solution would be in excess to the [free Ca24) that should be obtained. Thus, it is necessary to use Ca24 buffers that are made of a ligand in equilibrium with a metal, which in this case is Ca24.
Use of Fluorescent Tnc Derivatives and ‘Caged’ Compounds to Study Cellular Ca2+ Phenomena
Authors: Ashley C C , Griffiths P J , Lea T J , Mulligan P , Palmer, R E , Simnett Sarah J 
Published: November 2023

Abstract
1. Stncdanz as an indicator 1.2 Introduction Although it has been established using the photoprotein aequorin (Figure 1) that the free calcium concentration within muscle cells changes rapidly upon activation (1, 2) little is known of the time-course of calcium bound to the contractile system as a result of this transient change in the free calcium.
The Ca2+-Atpase of the Plasma Membrane
Authors: Carafoli Ernesto 
Published: November 2023

Abstract
1. Introduction The ejection of Ca2+ from cells is a necessity dictated by the existence in the plasma membrane of Ca channels which permit, albeit in a strictly controlled fashion, the penetration of Ca into the cell. Normally, Ca is about 10 000 times less concentrated in the cytosol than in the environment: therefore its ejection is an active process, and is indeed mediated by an ATP-powered pump. The pump was discovered by Schatzmann in 1966 in erythrocytes (1), and was subsequently found in a number of other eukaryotic plasma membranes (for reviews, see refs 2-5). The pump belongs to the P-class of ion-motive ATPases (6-7) and is now well-characterized functionally. More recent work on the purified pump has provided significant information on its structural properties (8).
Proteins Involved in the Uptake and Release of Ca2+ from the Sarcoplasmic Reticulum
Authors: Hamilton Susan L , Tate Charlotte A 
Published: November 2023

Abstract
1. Introduction Changes in the intracellular concentration of Ca2+ in skeletal muscle regulate the state of the muscle contraction and relaxation. A number of different proteins located in the sarcoplasmic reticulum (SR) are involved in the regulation of the Ca2+concentrations; some of the proteins with known roles and others with putative roles are listed in Table 1. In this chapter we will detail some of the methods used to study the two proteins most directly involved in controlling Ca2+ levels in skeletal muscle: (a) the Ca2+ ATPase, the protein responsible for the uptake of Ca2+ into the lumen of the SR, and (b) the Ca2+-release channel, the protein responsible for the release of the luminally sequestered Ca2+ into the cytoplasm.
The Role and Study of Mammalian
Authors: McCormack James G , Crompton Martin 
Published: November 2023

Abstract
The inner membrane of mitochondria of mammalian, and perhaps of all vertebrate, tissues contains Ca2+ transport systems that mediate continuous Ca2+ cycling across the membrane (Figure I; see refs 1-3 for reviews). The cycle comprises a Ca2+ uniporter which allows passive Ca2+ entry down the Ca2+ electrochemical gradient and one, or possibly two, active egress mechanisms. The principal efflux mechanism involves obligatory exchange between Ca2+ and Na+ with the probable stoichiometry of 2Na+: 1Ca2\ this exchange is linked to the chemiosmotic H+ circuit via Na+:H+ exchange. Mitochondria may also contain a Na+-independent mechanism for Ca2+ efflux, although whether this involves direct Ca2+: H+ exchange is controversial. Although cycling of Ca2+involves the dissipation of the protonmotive gradient, under normal cellular conditions the calculated overall energy cost is less than 1% of the respiratory capacity of the mitochondria (1, 2).
An enzymatic method for the complete mutagenesis of genes
Authors: Knowles Jonathan , Lehtovaara Päiyi 
Published: November 2023

Abstract
Site-directed mutagenesis has contributed considerably to our understanding of the relationship between protein structure and function. However, since site-directed mutagenesis is laborious, at least with regard to all the possible mutants that should be examined for a full understanding of any particular aspect, it is important that other more efficient methods for the investigation of protein structure by mutagenesis should be available. More importantly. site directed mutagenesis experiments are limited by the imagination of the investigator which has proved in recent years to be often too limited to understand the complexities of proteins.
Spiked oligonucleotide mutagenesis
Authors: Blacklow Stephen C , Knowles Jeremy R 
Published: November 2023

Abstract
Two approaches exist for the mutagenesis of cloned genes. In site-specific mutagenesis the presumed importance of a single amino acid is probed by changing or deleting it and examining the functional consequences. This approach is often helpful in providing experimental evidence for mechanistic suggestions developed from structural and kinetic studies. Random mutagenesis, on the other hand, does not rely on any preconceived notions based on structure or mechanism, but rather is employed to identify all possible changes that satisfy a particular functional selection. These two approaches are therefore complementary and both have been used in this laboratory for studies of structure function relationships in the glycolytic enzyme triosephosphate isomerase. In this chapter we describe a method for the random mutagenesis of any gene that has been cloned and expressed in Escherichia coli. Several different methods have been devised for the random mutagenesis of cloned genes (1). These include in vivo methods such as the use of mutator strains, in vitro enzyme-dependent approaches (e.g. ref. 2, and Chapter 8), and procedures that use chemicals to damage the DNA (e.g. ref. 3, and Chapter 7). However, it is known that mutagenesis by these procedures may result in an uneven distribution of mutations because of the presence of mutagenic ‘hot spots’ in the DNA. The method described in this chapter was developed to ensure complete randomness in the changes of nucleotides in the gene of interest.
Composites
Authors: Vincent Julian F V 
Published: November 2023

Abstract
Composite materials are composed of stiff strong fibres, plates, or particles (the discontinuous phase) in a relatively compliant matrix (the continuous phase). Composites with ceramic materials (CaCO3, hydroxyapatite, etc.) as the discontinuous phase are covered in Chapter 3. In other biological compo sites the discontinuous phase can be protein (as in skin and keratin) or polysaccharide (arthropod cuticle, plants). The chemical nature is not impor tant for mechanical analysis although bond energy and density are important. In order to provide reinforcement, the fibres have to be sufficiently long: maximum reinforcement is obtained with infinitely long fibres; collagen fibres in echinoderms have an aspect ratio of over 5000. The fibre takes forces from the matrix through shear forces developed at the fibre-matrix interface. This transfer of force occurs along a length of fibre determined by the relative moduli of matrix and fibre, the conditions at the interface, and the diameter of the fibre. In order to reinforce effectively, the length of the fibre has to be at least twice this transfer length. At lesser lengths the fibres reinforce less effectively, acting as filler particles. The aim in testing composite materials should be to integrate the morphology of the material (such as the amount, aspect ratio, and orientation of fibres) with the mechanical properties of the whole material and of the separate phases. Harris (1) and Piggott (2) are good introductions to definitions and analysis. Although not strictly composites, cellular materials have been included in this chapter because they are often constructed of composite materials (3).
Cartilage and joints
Authors: Smeathers J E 
Published: November 2023

Abstract
Cartilage is found in various forms in most invertebrates (Coelenterata, Annelida, Mollusca, and Arthropoda) and vertebrates; the literature on cartilage from a wide range of species has been comprehensively reviewed (1). It has been most thoroughly described for the vertebrates, hence this text refers exclusively to mammalian cartilage, the majority of experimental work having been done on human articular cartilage (2). Cartilage is the primary connective tissue of chordates. It can be modified into a variety of forms to produce a flexible coupling between bones and skeletal supporting structures. Vertebrae can move slightly because the fibrocartilagenous discs can bend, twist, and shear. The ends of long bones are coated with articular cartilage, which is very smooth and, when wetted with synovial fluid, allows the bones to slide easily over each other. Cartilage contains cells (chondrocytes) which produce a soft chondromucoid ground substance reinforced with collagen fibre network which can be considered as a composite material, exhibiting rubber-like properties.
Preparation and Handling of Biological Macromolecules for Crystallization
Authors: Lorber B , Giege R 
Published: November 2023

Abstract
In the crystallization of biological macromolecules the quality and quantity of the required material is important. Although experimenters may have no choice, difficulties in crystal growth may be linked to the nature or source of the biological material. Purification, stabilization, storage, and handling of macromolecules are therefore essential steps prior to crystallization. As a general rule purity and homogeneity are regarded as conditions sine qua non; however. quality of the macromolecules will depend upon the way they are prepared. Although some structures were solved with less than 1 mg of material (for example, ref. !), a few milligrams should be available for the first crystallization trials. Once crystals can be produced which are suitable for X-ray analysis, additional material will be needed to improve their quality and size and to prepare heavy-atom derivatives. For these reasons it is essential that isolation procedures arc able to supply enough fresh macro molecules of reproducible quality.
Seeding Techniques
Authors: Stura E A , Wilson I A 
Published: November 2023

Abstract
A seed provides a template for the assembly of molecules to form a crystal with the same characteristics as the crystal from which the seed originated. Seeding has often been used as a method of last resort rather than a standard practice. It has been used in situations when, after the first crystallization event. further attempts at crystallizing the same or subsequent batches of the same protein, under apparently identical crystalization conditions, either failed to yield any crystals at all, or provided crystalline precipitates or very small crystals. The use of seeding in crystallization can simplify the task of the crystallographer even when crystals can be obtained by other means. We will explore the various seeding techniques, and their applications in the growth of large single crystals.
Phase Diagrams
Authors: Ries-Kautt M , Ducruix A 
Published: November 2023

Abstract
Establishing a solubility diagram is one way of quantifying the influence of crystallization parameters (sec Chapter 1, Table 1) on the solubility of biological macromolecules. In this chapter, theoretical concepts of solubility will be developed firstly, to reiterate how a compound dissolves in a solvent, and to point out what i particular to biological macromolecules especially concerning the type of molecular interactions which can he found in a macromolecular solution. Indeed, interactions between solvent or additives and a biological macromolecule, which are necesary for its solubilization, can compete with intramolecular interactions responsible for the tertiary structure. The concepts of salting-in and salting-out will be summarized.
Aerodynamics of flight
Authors: Dudley Robert 
Published: November 2023

Abstract
Flying animals comprise the most morphologically diverse and numerically abundant form of life in the terrestrial biosphere. Animal flight can be described at a variety of phenomenological levels, ranging from the biophysics and ultrastructure of llight muscle to the behaviour of migrating locust swarms. An aerodynamic analysis of flight entails description of wing and body motions, evaluation of the forces associated with these motions, and calculations of the concomitant power requirements during locomotion. Additionally, the implications of wing and body morphology for more general measures of flight capacity, such as characteristic airspeed and manreuvrability, can be assessed quantitatively.
Isolation of sinoatrial node cells
Authors: Brown H F 
Published: November 2023

Abstract
The dissociation of cardiac tissue into single cells opened up new possibilities for physiological investigation which have led in the last decade to major advances in knowledge of heart function at the cellular level. Cardiac cell isolation was pioneered in the 1970s (1) and was quickly adopted by many other groups. The basic principle is simple: cardiac tissue is exposed to enzymes (always collagenase and often other enzymes such as trypsin, protease or elastase) in a Tyrode solution containing a very low concentration of calcium. Under these conditions, the cell junctions arc disrupted and will seal over to give intact but separate cells. In practice, it is not always easy to obtain cells that remain calcium tolerant when returned to physiological levels of calcium and that are neither too fragile nor too ‘tough’ for procedures such as patch–damp recording, while retaining their normal membrane pumps and receptors. Nevertheless, the isolation of ventricular and atrial cells is now successfully carried out in many laboratories.
Pharmacological analysis of synaptic transmission in brain slices
Authors: Henderson Graeme 
Published: November 2023

Abstract
1. Introduction
For the study of synaptic transmission in the mammalian central nervous system (CNS) the in vilro brain slice preparation presents a significant number of advantages over in vivo techniques.
(a) Foremost amongst these is the relative ease with which recordings can he obtained using extracellular, intracellular and whole-cell patch-clamp techniques.
(b) In certain slice preparations (e.g. the hippocampus), one can see under a low-power dissection microscope the presynaptic fibre tracts, the areas of synaptic contact and the cell body layer; thus, the stimulating and recording electrodes can be positioned at the desired site under visual control.
(c) Since the brain slice is maintained in an artificial bathing solution, the ionic composition of the extracellular environment can be manipulated.
(d) With intracellular and whole-cell patch-clamp recording, the ionic composition of the intracellular milieu can also be controlled.
(e) Drugs can be applied to the synapse under study in known concentrations by addition to the bathing solution.
Whole-cell patch-clamp recording from neurones in spinal cord slices
Authors: Pickering Tonye , Spanswick Dave , Logan Steve D 
Published: November 2023

Abstract
1. Introduction
The development of the patch-clamp methodology in the laboratories of the 1991 Nobel prize winners Neher and Sakmann has revolutionized our understanding of ion channels. The patch-damp recording technique was originally developed to allow high-resolution current recording from single ion channels (1). The vital step in this process is the formation of a gigaohm seal between the electrode glass and the membrane. This tight seal electrically isolates the few square microns of membrane in the pipette by reducing the leakage current to a level where picoampere currents flowing through single channels are resolvable.