bluuebunny/crossref_metadata_2025_split huggingface dataset:
PCR amplification of microdissected DNA
Authors: Johnson Daniel H 
Published: November 2023

Abstract
A common experimental strategy for understanding the molecular basis of developmental and physiological processes begins with the isolation of the genes encoding the cellular components which mediate these events. In many cases our knowledge of these genes is limited to the relative physical map location, on a chromosome, of mutant alleles which appear to perturb normal processes or contribute to disease states. With the advent of in vitro amplification synthesis of DNA (PCR) it has been possible to develop efficient protocols for the recovery of DNA restriction fragments from microdissected chromosomal regions harbouring specific genes of interest (1, 2).
PCR-directed cDNA libraries
Authors: Jane Sarah Jane , McPherson Michael J 
Published: November 2023

Abstract
Construction of a PCR-directed cDNA library from total RNA may provide the only methodological approach to analyse cell-specific gene expression where the amount of biological tissue is severely restricted. This approach is particularly applicable where a specific stimulus results in the particularly or differentiation of a small number of cells within a population. We are interested in studying changes in gene expression in plant roots where some 0.1 to 0.01% of the tissue is modified by an invasive pathogen.
Receptor-mediated endocytosis and lysosomal transport
Authors: Nolan Catherine M 
Published: November 2023

Abstract
Endocytosis is the process by which cells internalize their plasma membrane together with molecules adsorbed to the plasma membrane, fluid from the extracellular medium, and solutes dissolved in this fluid. Most cell types perform this process to greater or lesser extents. In addition to being a means of maintaining the correct proportion and composition of the plasma membrane, endocytosis can serve many other functions, including nutrition, regulation of the extracellular environment, antigen processing and presentation, and hormone processing. It may play an important role in processes such as growth and differentiation, wound healing, embryonic development, and pathological processes such as inflammation. The endocytic process can also be subverted by bacteria and viruses for penetration of them or their toxins into the host cell.
Network antibodies as tools for studying intracellular protein traffic
Authors: Vaux David 
Published: November 2023

Abstract
Many questions in cell biology can be reduced to a study of the interactions between two protein components, whether these are enzyme and substrate, surface receptor and extracellular ligand, or targeting signal and intracellular receptor. Many biochemical techniques can be applied to these problems, but there are occasions when these techniques are insufficient. Under these circumstances, it is justified to turn to an immunological approach to modelling the interaction between two proteins and to make use of the ability of the immune system to generate connected networks of interrelated immunoglobulins, network antibodies. In this chapter I will briefly outline the theoretical basis for this approach, consider the advantages and limitations of the method, and then provide detailed practical protocols.
Bioreactors for mammalian cells
Authors: Hanoa-Corrigan Anita 
Published: November 2023

Abstract
Contamination problems can occur at all stages of operation. At the early stages of expansion of a culture for inoculum preparation, operator errors and excessive dependence on anti-microbial agents contribute to increased risks of contamination. During larger-scale cultivation, problems with maintaining sterility can occur in a number of areas in the bioreactor including: exit and entry ports; air inlet and exhaust filters; valves and seals; bioreactor internals such as agitator shafts and probes; connectors and connecting lines to other vessels, pumps, analysers; etc. Successful sterile cultivation at any scale requires a combination of highly skilled cell culturists and well-designed (but not over-designed!) bioreactors. Care should be taken not to make a choice based solely on the bioreactor’s capability of achieving high cell or product yields. Some of the more sophisticated mammalian cell culture bioreactors are often prone to high contamination risks.
Conditions for density gradient separations
Authors: Dobrota Milosla V , Hinton Richard 
Published: November 2023

Abstract
While the theory of centrifugation may at first appear daunting, the actual techniques involved in density gradient centrifugation are in practice quite simple and can be mastered easily by a beginner. This chapter is written primarily for those new to the techniques of density gradient centrifugation and for researchers who find that there is no published method which gives the separation which they require and must develop their own. It is, however, rarely necessary to start completely from scratch as some useful pointers to approaches can be obtained from the numerous successful applications which have been developed already. As the authors know by experience, given the different properties of rotors, gradient media, and gradient design, one can spend one’s life trying to optimize separations rather than facing the real problem, the study of the properties of the separated particles. The aim of this chapter is to help people with the practical problems of designing gradient separation methods so that they can get on with real science. Hence the approach described is essentially empirical. However, theoretical considerations inevitably impose limitations on what can be achieved and it is those limits which will be addressed first.
Isolation and characterization of membranes and cell organelles
Authors: Evans W Howard 
Published: November 2023

Abstract
Centrifugal techniques have contributed crucially to the subcellular dissection of cells, providing important knowledge on their structure and function (1). Centrifugation is the basis of traditional methods for separating cell membranes and organelles. A number of other techniques which exploit various physical parameters (e.g. electrical charge) or biological properties (e.g. ligand affinity) have been investigated as ways of examining the complexity of organelles and membranes. However, even when using these other methods it is often the case that better results will be obtained if the material is first purified by centrifugal methods.
Centrifugation separations of mammalian cells
Authors: Brouwer Adriaan , Hendriks Henk F J , Ford Terry , Knook Dick L 
Published: November 2023

Abstract
Detailed analysis of the biochemistry, function, and behaviour of a specific cell population can be ascertained only on relatively homogeneous cell populations. However, cell preparations obtained from blood or tissues represent a mixture of different cell types. Many techniques are currently available for fractionating heterogeneous cell populations and for isolating fractions which are enriched for a specific type of cell, even if it constitutes only a minor component of the original population.
Matrix components produced by endothelial cells: type VIII collagen, SPARC, and thrombospondin
Authors: Sage E Helene , Bornstein Paul 
Published: November 2023

Abstract
The development, growth, and response to injury of the blood vasculature depend in part on interactions of vessel wall cells with their extracellular matrix (ECM). In addition to providing cues for other cells, the endothelial ECM directs many of the functions and responses of the endothelium itself. Since changes in endothelial cell behaviour (for example attachment, proliferation, biosynthetic activity, and/or migration) have been causally linked to one or more specific ECM macromolecules, identification of matrix-associated proteins secreted by endothelial cells has become critical to our understanding of vascular cell biology. Few proteins, with the exception of von Willebrand protein, have been shown to be specific to endothelial cells. Although several of them were either initially described or studied extensively as products of endothelial cells in vitro, extracellular proteins such as fibronectin, laminin, plasminogen activators and their inhibitors, various collagen types, thrombo-spondin (TSP), and SPARC, have been ascribed functions within a broad range of cells and tissues. Because of their potential role in the regulation of endothelial cell growth, three of these proteins have been chosen as subjects for this chapter: type VIII collagen, TSP, and SPARC.
Tenascin, laminin, and fibronectin produced by cultured cells
Authors: Erickson Harold P , Briscoe Gina 
Published: November 2023

Abstract
Tissue culture cells synthesize a variety of extracellular matrix (ECM) molecules and incorporate them into a matrix. In most cases, however, the incorporation into the matrix appears to be inefficient, and by far the majority of the ECM proteins are simply secreted into the medium as soluble molecules. One can harvest the ‘conditioned medium’ from confluent cell cultures every 2—4 days and purify the molecules of interest. If a monoclonal antibody is available in large supply, affinity purification on an antibody column is frequently the preferred method. A disadvantage of immunoaffinity purification is that the protein must be exposed to denaturing or chaotropic agents to elute it from the column. Alternative biochemical purification may be essential for proteins that are easily denatured, and is desirable whenever one is testing for unknown and potentially sensitive functions.
Sample preparation
Authors: Primrose William U 
Published: November 2023

Abstract
Over the last few years, there have been rapid and sustained advances in the hardware and software associated with the NMR analysis of biological macromolecules. Larger magnets, faster computers, and new and improved pulse sequences have been combined with isotopic labelling to make NMR the unrivalled method for answering biological questions at the atomic level in the solution state. These dramatic advances have perhaps tended to obscure the importance of correct preparation of the sample, which tends to be a rather poorly documented subject. The purpose of this chapter is to aid you in the correct choice of conditions for preparation of your vital and hard-won sample. This will then allow you to obtain the maximum benefit from the sophisticated spectroscopic methods described in other chapters in this book. The importance of correct handling of the sample cannot be over-emphasized, as no amount of expensive spectrometer and computer equipment can compensate for an intrinsically poor sample. A Formula One racing car will not break any lap records running on two-stroke fuel !
NMR Of Nucleic Acids; From Spectrum To Structure
Authors: Wijmenga Sybren S , Mooren Margret M W , Hilbers Cornelis W 
Published: November 2023

Abstract
Nucleic acids are perhaps the most intensively studied molecules found in nature, which is not too surprising in view of their role as carriers of genetic information. It has been a deep-rooted belief among molecular biologists that structure dictates function and, therefore, these studies have often involved the elucidation of structural aspects of these molecules, be it the linear base sequence or the three-dimensional arrangement of their nucleotide units. In a similar spirit it has been a deep-rooted belief among NMR spectroscopists that ‘ their ’ field of spectroscopy should be able to contribute to the elucidation of the structure and dynamics of molecules, in particular, biomolecules, in a way unparalleled by other forms of spectroscopy. Over the last few years this goal has come within reach thanks to the development of multidimensional NMR techniques which provide the necessary spectral resolution for the successful interpretation of the complicated spectra of biomolecules. Triggered off by the development of two-dimensional NMR, an avalanche of NMR studies on nucleic acids has appeared (1).
Structure Determination from NMR Data I. Analysis Of NMR Data
Authors: Barsukov Igor L , Lian Lu-Yun 
Published: November 2023

Abstract
This chapter deals with the strategies which are used to derive the experimental data required as inputs for calculation of the three-dimensional structure of a protein. The experiments used for spectral assignment described in Chapters 4 and 5 are also used for obtaining the data for structure calculations, although for the latter purpose, a more quantitative analysis is required. The principal structural data derived from NMR are scalar (J) couplings and NOEs, which are translated into restraints on dihedral angles and interproton distances, respectively. It is important to bear in mind that NMR leads to constraints on angles and distances, not to exact values of these parameters. The quality of the solution structure obtained has been shown to be proportional to the number of constraints used in the structure calculations.
Aetiology of type I diabetes: genetic aspects
Authors: Wassmuth Ralf , Kockum Ingrid , Karlsen Allan , Hagopian William , BäRmeier Heike , Dube Syama Lima , Lernmark Åke 
Published: November 2023

Abstract
Ancient literature has indicated that two forms of diabetes could be discerned. One was associated with emaciation, dehydration, polyuria, and lassitude, and the other with stout build, gluttony, obesity, and sleepiness (Bliss 1982). At the beginning of the twentieth century, however, and even after the discovery of insulin in 1921, diabetes was more often than not regarded as a single syndrome. A distinction was later made between juvenile and maturity onset diabetes mellitus but it was not until 1940 that these two forms of diabetes were suggested to be genetically distinct (Rotter et al. 1990). It also became clear that diabetes mellitus is not only insulin dependent or non-insulin dependent. A large number of disorders or conditions are associated with glucose intolerance; their causes range from mono genie disorders to diabetes secondary to a pancreatic affliction or to a peripheral insulin resistance induced by drugs or tumours. A recent survey lists more than 60 distinct genetic disorders associated with glucose intolerance and, sometimes, clinical diabetes (Rotter et al. 1990). Glucose intolerance and diabetes are therefore symptoms rather than defining diagnostic criteria. In order to properly diagnose insulin-dependent diabetes a set of diagnostic criteria are necessary. Such criteria.
Metabolic mapping of local neuronal activity
Authors: Kennedy Charles , Smith, Carolyn Beebe , Sokoloff Louis 
Published: November 2023

Abstract
The radioactive deoxyglucose method was developed to measure local rates of energy metabolism simultaneously in all components of the brain of conscious laboratory animals. It takes advantage of the extraordinary spatial resolution made possible by quantitative autoradiography (1). The dependence on autoradiography prescribed the use of radioactive substrates for energy metabolism, the labelled products of which could be assayed in the tissues by the autoradiographic technique. Although oxygen consumption is the most direct measure of energy metabolism, the volatility of oxygen and its metabolic products and the short physical half-life of its radioactive isotopes precluded measurement of oxidative metabolism by the autoradiographic technique.
Fluorescence—monitoring cell chemistry in vivo
Authors: Duchen Michael R 
Published: November 2023

Abstract
Many molecules, when exposed to light of appropriate energy, absorb photons which excite electrons into a higher energy state. As those electrons relax to their original orbits, photons are re-emitted, a phenomenon known as fluorescence. The detailed spectra of the light that is absorbed and emitted may vary accordinging to other interactions of the molecule-its binding to Ca2+, protons, or other ions, the electrical field it senses, and so on. In recent years, we have seen major developments in fluorescent probes that can transmit information about intracellular events within living cells. The changes in intracellular concentrations of ions and other second messenger systems that play so fundamental a role in cell function have at last become accessible to direct study without having to impale cells with microelectrodes.
Enzyme reactions
Authors: Crabbe M J C 
Published: November 2023

Abstract
This chapter introduces some of the uses for microcomputers in studies of enzyme reactions. In many ways, it follows on from an earlier chapter on the applications of microcomputers in the analysis of enzyme reactions, published in this series in 1984 (1). The intervening years have seen great advances in the facilities offered by microcomputers. Machines with 20 MHz 32-bit CPUs and 4 Mbyte RAM are now commonplace, while in 1984 laboratories were more likely to be using micros with 4 MHz 8-bit CPUs and 32 kbyte of RAM. This increase in speed and storage facilities, not to mention improvements in screen resolution and colour display, have made microcomputers an invaluable tool in any laboratory. Similar advances in networking, laser printing, and X-Y plotting now enable raw data to be manipulated directly into publishable hard copy.
Gas Chromatography
Authors: Baugh P J 
Published: November 2023

Abstract
Gas chromatography (GC) has been one of the most used and successful chromatographic techniques since its inception over 40 years ago and is employed by almost every type of chemical and related industry in research and development, quality control and specifications, environmental monitoring —personnel, and process chemistry, biochemical and biological research establishments, clinical and toxicological departments of hospitals and related institutions, analytical and consultant/public analyst laboratories, water companies and regulatory bodies (National Rivers Authority), and higher education teaching/research institutions at all levels.
Applications of packed and capillary GC
Authors: Baugh Peter J 
Published: November 2023

Abstract
Although it can be said that capillary columns (high performance, HP) have superceded packed columns0 (low performance, LP) in GC applications and industrial uses of GC, the latter for many years has remained the preferred technique for quality control and chemical process monitoring. The reason has not necessarily been a reluctance to change but purely because LP columns could, with high capacity, achieve the performance adequately enough to satisfy the requirements. In such applications it was only necessary to determine the major components and impurities (&lt; 1%) could be ignored.
Synthesis Of Oligo-2’-Deoxyribonucleoside Methylphosphonates
Authors: Miller Paul S , Cushman Cynthia D , Levis Joel T 
Published: November 2023

Abstract
Oligo-2'-deoxyribonucleoside methylphosphonates contain nuclease resistant, non-ionic internucleotide bonds. These oligomers are taken up intact by mammalian cells in culture. In addition to their unique physical properties, these oligonucleotide analogues give sequence-specific inhibitory effects when targeted against specific messenger RNA or precursor messenger RNA in mammalian cells in culture. The use of oligonucleoside methylphosphonates as antisense reagents in cells has been extensively reviewed (1-3). This chapter will describe the automated synthesis, deprotection, and purification of these oligomers, methods used to characterize the oligomers, and a general description of procedures for preparing them for use in cell culture experiments.
Oligonucleotides With Reporter Groups Attached to the 5’-Terminus
Authors: Sinha Nanda D , Striepeke Steve 
Published: November 2023

Abstract
Traditionally, oligonucleotides or DNA have been labelled and detected via use of P-labelled ATP and enzymatic phosphorylation. Although of high sensitivity, this technique has intrinsic hazards, expense, and problems associated with the short half-life of this isotope. Recent advances have shown that high sensitivity detection can be achieved by alternative markers based on fluorescent, chromophoric, or chemiluminescent detection. Incorporation of these labels can be achieved by enzymatic (1-8) or chemical (9-23) methods. One of the simplest and most useful methods involves introduction of primary amino or sulphyrdryl (thiol) groups to the 5’-terminus of the chemically assembled support-bound oligonucleotide (14-23). Subsequently fluorophores, chromophores, biotin, or alkaline phosphatase may be added. Further examples are discussed in Chapter l0.1With this approach the position of the amino or thiol group is defined and unambiguous, and there is negligible interference in subsequent hybridizations. Additionally, the 3’ -terminus of the oligonucleotide chain is free for enzymatic manipulations as required in Sanger’s sequencing and PCR amplification applications (14, 24-26). The attachment of other groups to the 5’-terminus of oligonucleotides is described in Chapter 12.
The European Community at the cross-roads
Authors: Vilar Dr Emilio Rui 
Published: November 2023

Abstract
When I have to address an audience like this one, I always remember the story of a famous speechmaker, who was able to be at the same time brief and accurate. Invited to speak about the great problems of mankind before a huge audience, he said: “The major problems of mankind are ignorance and indifference” and became silent. Among the audience someone asked his neighbour: “What did he say?” “I don’t know and I don’t care” was the reply.
Improvement of malting barley quality
Authors: versen John Ing 
Published: November 2023

Abstract
The current trend in one of the major malting barley growing areas of the world—Central Europe, is that more and more feed barley is grown on expense of especially row spring barley types. To revert this development and to breed even better malting barley for the future a research programme to “Adapt Barley to Industrial Needs”—(ABIN) was initiated in 1988. Within the ABIN project the most modern techniques are employed to solve some of the problems which faces the maltster and brewer, when present day barley is used as raw material in the industry. The lecture will present the background, overview and current status of the ABIN project.
Germination profile—a new term in barley analyses
Authors: Riis Preben , Bang-Olsen Kirsten 
Published: November 2023

Abstract
Production of high quality malt requires that the barley germinates fast, completely and uniformly. Based on an extension of the BRF method to measure Germlnatlve Energy, we propose two supplementary measures of quality: Germination Index, a measure of germination rate, and Germination Homogeneity, a measure of the variance of germination. To evaluate the feasibility of these measures, series of micromaltings were performed using barley with artificially reduced vigour. The results of these micromaltings show that the three measures for germination, for which we use the unifying term Germination Profile, can be a valuable tool in controlling maltings.
Untersuchungen zur Wasseraufnahme und Wasserverteilung der Gerste auf der Grundlage von rasterelektronenmikroskopischen Analysen (REM) der Endospermstruktur
Authors: Rath F , Schildbach R 
Published: November 2023

Abstract
Precise knowledge of the physiology of water uptake and the factors influencing it, is a requirement for the exercise of influence achieved during the malting process and also for the development of new criteria for selection in barley breeding.
Sulphur compounds in brewing
Authors: Clarke B J , Burmeister M S , Krynicki L , Pfisterer E A , Sin Y O , Sime K J , Hawthorne D B 
Published: November 2023

Abstract
Routine monitoring with high sensitivity of the concentration of a range of volatile sulphur compounds in beer has now become feasible. With the development of the chemiluminescent detector the ready quantitation of at least six sulphur compounds in a single analysis is possible, the major components detected are dimethyl sulphide and hydrogen sulphide.