bluuebunny/crossref_metadata_2025_split huggingface dataset:
The future — towards effective prevention and cure for myotonic dystrophy
Authors: Harper Peter S 
Published: November 2023

Abstract
There is no proven medical treatment that influences the rate at which myotonic dystrophy deteriorates or which can prevent its onset in those carrying the genetic change. There is a great deal of research that is ongoing into myotonic dystrophy. Now that the genes responsible for myotonic dystrophy have been identified, the focus of new research is on finding effective treatment.
Introduction
Authors: Wright Michael , Hamzah Ednin , Phungrassami Temsak , Bausa-Claudio Agnes 
Published: November 2023

Abstract
Southeast (SE) Asia is divided into two sub-regions. Mainland Southeast Asia comprises countries such as Cambodia, Laos, Myanmar, Thailand, and Vietnam. Maritime Southeast Asia encompasses the island states of Brunei, Indonesia, the Philippines, Singapore, and Timor Leste. Malaysia, with two states on the island of Borneo and the others on the Malay Peninsula, straddles the two (Fig. 1.1).
Epistemic Humility, Arguments from Evil, and Moral Skepticism
Authors: Howard-Snyder Daniel 
Published: November 2023

Abstract
The conclusion is frequently more nuanced: “it is very likely that there is no such reason’’ or “more likely than not” or “more likely than it otherwise would be”. Some critics reject the premise: we do see how some reason would justify God. These are the theodicists. Others accept the premise but reject the conclusion: the evidence or non-evidential warrant for God’s existence is much better than the evidence for no justifying reason.
Evaluating Religion
Authors: Kapitan Tomis 
Published: November 2023

Abstract
Controversies about religion, the value of religious practices, the grounds for religious beliefs, or the benefits and dangers of religious institutions, are probably as old as religion itself. The disagreements are usually vivid and longstanding; apart from politics, few sectors of human civilization can rival religion in division, acrimony, and bloodshed. Criteria for rationally assessing these debates are desirable, therefore, and to provide them is the object of this chapter. But little headway can be made on this matter without first circumscribing the topic. Just what is “religion”? What makes something “religious”? How extensively can these concepts be applied? Without answers to such questions, we cannot be sure how broad a sweep is made by a given critique or defense of religious phenomena.
Ockhamism and Molinism—Foreknowledge and Prophecy
Authors: Warfield Ted A 
Published: November 2023

Abstract
On earlier occasions I have argued for the compatibility of human freedom with divine foreknowledge and defended that argument from criticisms. On those occasions I provided and defended only my positive argument for my compatibilist position. I did not reply to standard arguments for the incompatibility of human freedom and divine foreknowledge. Nor did I say anything substantive about the “Ockhamist” approach to the foreknowledge issue thatI favor.
Radioisotope Detection Using X-Ray Film
Authors: Laskey Ron A 
Published: November 2023

Abstract
During the last two decades gel electrophonsis has steadily replaced other methods for analytical fractionation of macromolecules. This chapter describes and discusses methods of detecting radioisotopes within flat samples such as acrylamide or agarosc gels, nitrocellulose t1ltcrs, thin layer chromatography plates and paper chromatograms. For all of these samples X-ray film provides a convenient non-destructive means of isotope detection which combines high sensitivity with high resolution.
Strategy and Tactics in Receptor Binding Studies
Authors: Hulme E C , Birdsall N J m. 
Published: November 2023

Abstract
The questions answerable by receptor-binding studies fall into two major categories: Firstly, there are fundamental questions which one may ask about: the molecular pharmacology of receptors, the mechanism of ligand binding, and the nature and mechanism of receptor-effector in1eractions. Secondly, there are applied questions which concern the cell biology and anatomical distribution of receptors, particularly: their concentrations in different tissues, their distribution on different eel! types, their ontogenetic development, and how their synthesis, membrane insertion, coupling, degradation. and re cycling are regulated in a given cell! type.
Peptide mapping and the generation and isolation of sequenceable peptides from receptors
Authors: Wheatley M 
Published: November 2023

Abstract
As increasing numbers of receptors are purified to homogeneity, research interests are aimed at elucidating structural information about the receptor proteins themselves. This encompasses obtaining general sequence information often used for constructing the probes required for cloning the receptors, and in addition investigating the location within the molecule of functionally important amino-acid residues or moieties. A popular approach to this is site-directed mutagen ésis of a residue or region of interest, followed by expression of the protein and its characterization. By virtue of its indirect approach, it is sometimes difficult to relate any observed change in function with a given mutation. A complementary and synergistic approach is to chemically modify the pure receptor directly, and then analyse the protein by peptide-mapping studies and protein-sequencing techniques. The aim of this chapter is to provide the information necessary for peptide mapping receptors utilizing the low-technology, and therefore relatively cheap, high-resolution system of sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Furthermore, information will be presented pertinent to the preparation and purification of receptor-derived peptides suitable for sequencing. It is not within the scope of this chapter to describe the actual sequencing procedure itself. When appropriate, examples from our research on muscarinic acetylcholine receptors (mAChRs) will be presented.
IL-2 as a short-range physiological mediator
Authors: Mitchison N A 
Published: November 2023

Abstract
It is now generally accepted that the immune system neither could nor does operate as a perfect gas, in which cells, antigens, and cytokines move at random. On the contrary, movement is under tight constraint, mainly as a result of adhesion between cells. Adhesions, plus the polarized secretion to which they give rise, control the site of action of cytokines, and make these molecules function largely as ultra-short range signal transmitters. In this respect cytokines differ profoundly from classical hormones in their systemic activity, although the boundary is blurred by agents such as the pituitary releasing hormones which act more or less locally. A comparison with neurotransmitters would be more apt, and the promiscuity of these molecules echoes the promiscuity found among the cytokines of the immune system. Consideration of neurosecretion should remind immunologists that long-range and short-range transmitters probably evolve from one another, and that the arrangement of cytokines as it occurs in man should be regarded as a snapshot rather than a stable picture. Furthermore, some of the molecules that transmit information between cells may have started their evolution inside cells; purinergic transmission may be a case in point.
Importance of steroidogenesis in specific hormone action
Authors: Funder John W 
Published: November 2023

Abstract
Six classes of hormonal steroids are currently recognized in mammalian systems – mineralocorticoids, glucocorticoids, androgens, oestrogens, progestins, and vitamin D. This classification was made historically on effector grounds – growth of a cock’s comb for androgens, hepatic glycogen deposition for glucocorticoids– long before anything was known of receptors or the subcellular mechanism of steroid action. In invertebrates, ecdysones are clearly an additional class of physiological steroid hormone, and in a variety of species both endocrine and paracrine (neurosteroid) roles are currently being explored for a number of steroids (e.g. dehydroepiandrosterone, pregnenolone). What remains true, however, is that of the hundreds of different steroids in various body compartments a (patho)physiological role as a signal is assigned to fewer than a dozen, with the rest being considered as precursors, biosynthetic intermediates, or degradation products.
Nuclear hormone receptors as transcriptional activators
Authors: Gronemeyer Hinrich 
Published: November 2023

Abstract
Multicellular organisms have established sophisticated techniques of information transfer between cells and body compartments. The information that is transmitted can be highly complex and can result in the alteration of gene programmes involved in cellular differentiation, proliferation, or reproduction. Despite the complexity of these events, the inducing signals can be relatively simple molecules, such as small peptides or cholesterol derivatives. Apparently, the need for a high speed of information transfer means that the signals must be small, highly diffusable molecules. However, only limited structural information can be stored in the chemical structures of, for example, steroids, thyroids, or retinoids, and therefore the interpretation of the signal is of major importance and requires a highly specialized system that specifies the appropriate gene programmes. This interpretation can be based on signal-specific nuclear receptors that are expressed in target cells – those cells that are genetically determined to respond to the signal. Understanding the molecular mechanisms by which the information stored in the signal is converted into gene programming is one of the exciting areas of molecular biology.
Repression of transcription by nuclear receptors
Authors: Drouin Jacques 
Published: November 2023

Abstract
We now have a fairly detailed understanding of the mechanisms by which nuclear receptors recognize their target genes and activate transcription (Chapters 3 and 5). Much less can be said about the ways in which these receptors repress transcription of other target genes. At the onset, it is interesting to note that early studies of the action of steroids on their target tissues indicated, for example, that glucocorticoids decrease the level of about the same number of liver proteins as they stimulate. In all likelihood, many of these changes will be ascribed to transcriptional regulation.
Mammalian Cell-Cycle Analysis
Authors: Darzynkiewicz Zbigniew 
Published: November 2023

Abstract
A plethora of methods designed to probe the cell cycle of mammalian cells has been developed in the past several decades. The scope of this chapter makes it impossible to fully describe them. Therefore, the essential features of the most useful techniques are presented. Outlines of particular protocols, virtues, drawbacks, and applications of the methods, and practical notes regarding critical steps, are given. To master these techniques, the reader is encouraged to consult the articles which describe them more extensively, and which arc referenced throughout the text. The aim of this chapter, therefore, is to introduce diverse techniques, which may be of help in selecting the most appropriate approach for a particular task.
Cell-Cycle Analysis Using The Filamentous Fungus Aspergillus Nidulans
Authors: Oakley B R , Osmani S A 
Published: November 2023

Abstract
The filamentous fungus Aspergillus nidulans has proved to be an excellent experimental system for the identification, isolation, and analysis of genes that are essential lo cell-cycle progression and regulation. Studies in A. nidulans have led to the discovery of several key elements of cell-cycle progression and regulation that were not first identified in other experimental systems (1). It is a homothallic ascomycetous fungus that has both a sexual and an asexual life cycle (Figure 1). The classical genetics and molecular genetics of A. nidulans have been previously described (2) and are, in many respects, as advanced as the yeast systems. his the purpose of this chapter to describe some of the techniques that can be utilized to study the cell cycle in A. nidulans.
Analysis Of Dna Replication Origins And Directions By Two-Dimensional Gel Electrophoresis
Authors: Huberman Joel A 
Published: November 2023

Abstract
Prior lo 1987 when the 2D gel electrophoretic ‘replicon mapping’ techniques were inlroduced (1, 2), no generally applicable method was available for the identification or characterization of cukaryotic DNA replication origins. Consequently, it was uncertain whether or not specific origins existed. Thanks to new results obtained with the 2D gel techniques (and, more recently, with additional origin mapping methods, refs 3-5), a great deal of information about eukaryotic origins is now available. Several recent reviews discuss the biological importance of these new results (6--9).
Preparation of DNA for use as probes
Authors: Byrnes Lucy , Enenkel Barbara , Gannon Frank , Smith Terry 
Published: November 2023

Abstract
The primary goal of this chapter is to describe the preparation of DNA for use as probes. The preparation of RNA probes and oligonucleotide probes are covered in Chapters 7 and 5, respectively. Preparative methods to obtain a specific DNA fragment from either a plasmid or phage which contains it are initially described. The generation of mixed DNA probes corresponding to the messenger RNAs present in a cell is the next general topic covered in this chapter. Finally the chapter describes the preparation of adequate target material for DNA hybridization and detection using probes.
Homogenization of tissues and cells
Authors: Graham John M 
Published: November 2023

Abstract
Many investigations into the structure and function of cells and tissues require, at some stage, the isolation of a particular membrane or subcellular organelle. The successful isolation and fractionation of subcellular membranes from any source, from animal cells, plant cells, or microbial organisms depends critically on the ability to disrupt the cell in such a way that the subcellular particle of interest can be isolated in high yield and as free from contaminating structures as possible. Moreover, the isolated biological particle should retain as much of its normal structural and functional integrity as possible and be in a form which is amenable to the analytical techniques to be used subsequently.
Isolation and characterization of peroxisomes
Authors: Völkl A , Baumgart, E , Fahimi H D 
Published: November 2023

Abstract
Peroxisomes (PO) are organelles ubiquitously distributed to most pro- and eukaryotic cells (1, 2). Depending on the cell type and the sets of enzymes which they contain, synonymous designations such as glyoxisomes or glycosomes are used with ‘microbodies’ as the all-embracing term. Mostly, PO are spherical or spheroidal particles with a diameter between 0.2–1.0 µm. In some tissues, however (3–6), they may be irregularly shaped.
PFGE in the study of a bacterial pathogen (Haemophilus Influenzae)
Authors: Hood D W 
Published: November 2023

Abstract
Bacterial chromosomes are relatively small and much simpler to analyse than the larger, more complex, genomes of higher eukaryotic organisms. Pulsed field gel electrophoresis (PFGE) is capable of resolving DNA molecules up to ten million nucleotides in length which should enable one to view the complete genome of most bacteria as a single electrophoretic band. Restriction endonuclease digestion of bacterial genomic DNA and resolution of the resulting fragments by PFGE has facilitated the construction of physical maps of bacterial chromosomes. Since the first bacterial chromosomal restriction map was determined for Escherichia coli (1) these have now been obtained for an ever-increasing number of bacterial species. They are proving to be extremely useful in studying many aspects of the genome and genetic and molecular studies on chromosome organization and function can now be carried out directly on intact chromosomes. In particular, for the subject of this chapter, genomic maps have enhanced our investigation of the genetic organization of pathogenicity determinants in the bacterium Haemophilus influenzae. These studies will serve to illustrate various aspects and uses of pulsed field gel electrophoresis relevant to the analysis of bacterial chromosomes.
Measurement of inositol phosphate turnover in intact cells and cell-free systems
Authors: Shears Stephen B 
Published: November 2023

Abstract
The question as to whether a particular stimulus activates PLC activity in intact cells can be answered by demonstrating either that levels of the inositol lipid substrates decrease (see Chapter 1), or that there are increases in the levels of Ins(l,4,5) P 3 (see below and Chapter 8) and/or 1,2-diacylglycerol. However, more detailed knowledge of the rate of PLC activity will not be obtained from these specific assays alone. For example, one of the consequences of receptor occupation is a stimulation of inositol lipid resynthesis to ensure a compensatory supply of substrate to ensure continuing PLC activity (3).
Mass assay of inositol and its use to assay inositol polyphosphates
Authors: Balla Tamas 
Published: November 2023

Abstract
Myo-inositol was discovered in 1850 as a component of muscle tissue, but its importance was not recognized until about 100 years later when deprivation of dietary inositol in various animals was shown to cause disorders including alopecia and lipodystrophy in liver and intestinal epithelial cells (see ref. 1 for review). These changes could be reversed by administration of inositol and increased in severity when the intestinal flora was destroyed, since inositol is probably produced by intestinal microorganisms. Furthermore, several mammalian cell lines have been found to require inositol to sustain growth (2), although some cells can grow without added inositol because they can synthesize sufficient amounts to support their needs (3).
Preparation of RNA:protein complexes for X-ray crystallography and NMR
Authors: Price Stephen R , Oubridge Chris , Varani Gabriele , Nagai Kiyoshi 
Published: November 2023

Abstract
Understanding the molecular mechanism of RNA recognition by protein requires knowledge of the three-dimensional structure of RNA:protein complexes through application of X-ray crystallographic and NMR spectroscopic techniques. To date, much of our knowledge of this recognition is derived from studies of complexes of tRNA and aminoacyl-tRNA synthctases (Gin, Asp, Ser. and Lys) (1-4) or elongation factors (5).
Proteolysis Of Native Proteins As A Structural Probe
Authors: Hubbard Simon , Beynon Robert J 
Published: November 2023

Abstract
A common use of proteinases in vitro is the degradation of proteins to limit peptide mixtures in which all susceptible bonds have been cleaved (Chapter 8). To facilitate this process, the substrate would usually be first denatured, by heat, urea, guanidinium hydrochloride, by precipitation with trichloroacetic acid or similar treatments. As a result of the destruction of the higher-order structure in the substrate, all susceptible (i.e. in accordance with the primary cleavage specificity) bonds will be attacked rapidly and with rates that, compared with native state digestions, are approximately similar and influenced only by local changes in subsite occupancy; in this way, the peptide mixture limit is rapidly generated. The need for the prior denaturation step implicitly assumes that retention of native structure will slow or modify the proteolytic attack, and this is indeed the case. The role of three-dimensional structure of a protein substrate in modification of extent and rapidity of proteolysis is incompletely understood, but a recurrent observation is that proteolysis is more limited, both in the extent and in the rate of hydrolysis. This restriction of proteolytic attack on native proteins also underpins most proteolytic interactions of physiological relevance, and thus, the study of limited proteolysis is also relevant to processes in vivo.
Electron microscopy of viruses
Authors: Gelderblom H R , Biel S 
Published: November 2023

Abstract
Though occasionally considered as laborious, old-fashioned, and unnecessary, the detection and identification of viruses and other agents by electron microscopy (EM) can make an important contribution to many basic and applied biomedical routine procedures. In some fields, for example, vaccine production, EM even forms an integral part of the production controls (Table 1). While the majority of direct and indirect virus detection techniques (e.g. ELISA, PCR) are highly sensitive, but also highly specific for just one individual species or family of agents, EM works differently. Based on its undirected approach and its high resolution, EM is able to detect routinely and characterize morphologically all structures greater than 15 nm in diameter, which fortunately coincides with the size of the smallest viruses (1).
In vitro replication of RNA viruses
Authors: Banerjee R , go M , Izumi R , Datta U , Dasgupta A 
Published: November 2023

Abstract
RNA viruses are very diverse and utilize many distinct strategies for replicating their genomes. The genome organization of these viruses vary widely, thus the translation and replication strategies employed must also be varied. However, despite all the complexities of RNA viruses and their differing replication strategies, there are many similarities at the molecular level (1). Some of the common in vitro techniques employed to study these RNA replication strategies as well as methodologies to study in vitro RNA-protein interactions will be discussed in this chapter.
Interaction of standard mechanism, canonical protein inhibitors with serine proteinases
Authors: Laskowski Michael 
Published: November 2023

Abstract
Proteinases have a cradle-to-grave relationship with proteins. They assist in the birth of the proteins by removing the initiating methionine (Met) residues. They further aid in delivering them to an appropriate destination by removing the signal peptides. At death, they convert both exogenous proteins (food digestion) and endogenous proteins (protein turnover) to amino acids, which are then utilized for new protein synthesis. However, the most striking is the interaction of proteins with proteinases in their adult life. Protein processing turns numerous activities on or off. These activities in turn are responsible for a large range of biological phenomena such as blood clotting, clot dissolution, protein hormone action, emergence of the silk moth from cocoons, penetration of outer layers of ova by sperm, differentiation, cell death, and apoptosis (1). Along with so many important functions there are also many dangers associated with proteolysis. It must be rigidly controlled in time and place in order to be effective. Clotting from head to toe from a cut or digesting one’s own pancreas rather than that of one’s victim would be just a few untoward examples of uncontrolled proteolysis.