bluuebunny/crossref_metadata_2025_split huggingface dataset:
Radioisotopes and Immunoassay
Authors: Gearing Adrian F Bristow. Andrew J H , Thorpe Robin 
Published: November 2023

Abstract
Immunmissay is the quantitative measurement of a substance of interest (‘analyte’) using antibodies which bind specifically to that analyle. For the re t of this chapter, these two main components of the assay will be referred to as ligand and antibody respectively. In order to perform such an assay, it is necessary to measure the binding of the ligand to the antibody at very low concentrations and this has most commonly been achieved with the use of radiosotopes. The first practical application of the principle was the technique of radioimmunoassay, invented by Yalow and Berson in the late 1950s (1), for which the Nobel prize was subsequently awarded. This technique, and subsequently developed related techniques such as immunoradiometric assays and enzyme-linked immunosorbent assays, have revolutionized the biological sciences. Using these methods the minute quantities found in biological materials of a whole range of analytes such as protein hormones, steroids, viral antigens, cytokincs, and clotting factors can be accurately measured without relying on difficult and sometimes imprecise measurements based on their biological activities. Indeed, it is hard to overestimate the impact that the development of immunoassays has had on the life sciences.
Federal Regulations on Use of Radionuclides in the United States of America
Authors: Nelson Neal S 
Published: November 2023

Abstract
In 1967, the World Health Organization reported that regulation of devices and materials emitting ionizing radiation was divided among fifty state governments, several city governments, six Federal agencies, a Federal Council and a Federal Commission (1). In a 1978 review, five Federal agencies and twenty-five states were reported to be regulating some aspccb of radiation protection and control (2). Unfortunately the situation in the 1980s is not much improved.
Dopamine receptors: isolation and Molecular Characterization
Authors: Strange Philip G , Williamson richard A 
Published: November 2023

Abstract
The actions of the catecholamine neurotransmitter dopamine can be accounted for by the binding of dopamine to D1 and D2 dopamine receptors. The properties of the two receptor classes have been discussed in detail elsewhere (1,2) and they appear to be distinct proteins with different pharmacological properties linked to different biochemical and physiological events. The isolation and purification of the receptor proteins is essential in understanding how the two receptor systems function. These methods will further enable the composition of the two proteins to be determined, will enable studies on receptor reconstitution to be performed, and will assist in the cloning of the genes for the receptors. Even when the genes have been cloned and the primary structure of the receptors has been obtained, studies on the isolated proteins will be required in order to locate key regions of the receptor, e.g. the ligand-binding site. This underlines the need for methods of purification.
Measurement and control of intracellular calcium
Authors: Gurney Alison M 
Published: November 2023

Abstract
Calcium is an important intracellular messenger in transducing responses to activation of many receptor types. Its concentration is tightly regulated in cells, with most of it bound up in intracellular stores and only a small fraction freely available in the ionized form. In order to understand how Ca 2+ concentration is regulated in cells, as well as its role in transducing receptor signals, it is clearly important to know how the concentration of the free ion changes in response to various stimuli, and how cells respond when intracellular Ca 2+ is changed.
Phosphate-labelling studies of receptor tyrosine kinases
Authors: Mahadev Louis C , Bell john C 
Published: November 2023

Abstract
The extensive literature on receptor tyrosine kinases and the intense research effort it currently receives reflects its central role in the regulation of mitosis, aberrations of which result in uncontrolled proliferation of cells and the formation of tumours. This literature is comprehensively summarized and reviewed in several recent articles and is not to be considered here (1 3). This chapter deals with the practical aspects of demonstrating that a particular protein or protein complex is a receptor tyrosine kinase (RTK) which is defined as one in which the binding of a specific ligand results in the direct activation of tyrosine kinase activity intrinsic to the receptor. It is obvious immediately that there are two criteria that must be satisfied. First, the putative receptor must be shown to have the appropriate ligand-binding characteristics. Approaches to demonstrat ing respectable ligand binding (summarized in Section 2.1) are considered in detail in a companion book in this series (25) and are not described here, although a brief outline is given in Appendix 1.
A reconciled view of the non-MHC requiring cytotoxicity phenomenon
Authors: Triebel F , Hercend T 
Published: November 2023

Abstract
Non-MHC requiring cell-mediated cytotox1c1ty is now recognized as an important effector function generated during cellular immune responses. Many studies over the years have contributed to delineate both the different cell subpopulations and the membrane molecules involved in these cytotoxic reactions. Among human peripheral blood lymphocytes (PBL), two types of effector cells are now well defined: the natural killer (NK) cells and the activated cytotoxic T lymphocytes (CTL), wither TCR α/ β+or γ/ δ+, which mediate some non-MHC requiring killing.
Specificities involved in recognition of tumour target cells by IL-2 activated NK cells and T cells; ‘one receptor’ versus ‘multiple choice’ models
Authors: Kärre K 
Published: November 2023

Abstract
The specificity of cytotoxic activities associated with in vivo or in vitro administered IL-2 is usually discussed in terms of LAK activity. Without further specification, the concept of LAK cells (or LAK activity) may be misleading. Studies in recent years indicate that IL-2 administration not only can activate different effector cells, but that some of these can also use alternative recognition strategies. The discussion in this paper will therefore take into account different principles for cell mediated recognition that have emerged in the literature in recent years. This will include a multiple choice model for NK cells as well as the paradigm of peptide presentation by MHC molecules to CTL. The basis for the discussion of different effector cells and the effect of low dose versus high dose IL-2 will be introduced by a brief overview of the historical development in interpretations of lymphokine activated killing of tumour cells.
Involvement of cytokines in combined modality preclinical approaches to cancer treatment
Authors: Wiltrout R H , Hornung R L , Futami H , Sayers T J , Young H A 
Published: November 2023

Abstract
About 15000 to 20 000 new cases of renal cell cancer are diagnosed each year in the United States. Renal cell cancer is relatively resistant to most chemotherapeutic drugs, which makes it a candidate for alternative treatment approaches such as immunotherapy in combination with chemotherapy. The rationale for believing that such an approach may prove useful is based on the fact that primary renal cancers are often fairly vascularized and responses to biological response modifiers (BRM) such as interferon-0( (IFN-0() and interleukin-2 (IL-2) have been documented.1−4 These observations suggest that preclinical animal models for renal cancer could be useful for testing the antitumour activity of BRMs, either alone, or in combination with chemotherapeutic drugs or other BRMs. The model chosen for preclinical study in our laboratory is the Renea adenocarcinoma of BALB/c mice. The Renea tumour arose spontaneously and its growth and progression in syngeneic mice has been previously characterized. The tumour spontaneously metastasizes from an intrarenal (IR) implant to the regional lymph nodes, lungs, liver, and spleen, as well as some other organs.5,6 The immunogenicity of Renea has been determined to be relatively low.
Chromatin structure and position effects
Authors: Kipling David 
Published: November 2023

Abstract
In Saccharomyces cerevisiae telomeric DNA sequences can influence the expression of genes located close to them, and may also influence the timing of replication of adjacent genomic regions. Genetic analysis has revealed that this position effect on gene expression is closely related to the transcriptional repression that occurs at the silent mating-type loci. Because the latter has been characterized in great detail it will be described first, thus enabling telomere position effects to be reviewed in context.
Ribosomes and polysomes
Authors: Bommer Ulrich A , Burkhardt Nils , Junemann Ralf , Spahn Christian M T , Triana-Alonso, Francisco J , Nierhaus Knud H 
Published: November 2023

Abstract
Isolation of ribosomes and assay systems for testing the translational apparatus have been part of the experimental routine of many laboratories for more than two decades, and excellent collections of the methods have been published previously, including two books in this series. However, a number of methods have been gradually and continuously improved and optimized. Major developments have included the improvement of ribosome isolation procedures, the establishment of highly efficient assay systems, and the application of heteropolymeric mRNA. The protocols in this chapter are concerned with the isolation of polysomes, ribosomes, and ribosomal subunits from prokaryotic and eukaryotic sources, as well as test systems for both total protein synthesis and single ribosomal functions, taking into account the recent developments.
RNA-protein interactions in Rnase P
Authors: Gopalan Venkat , Talbot, Simon J , Altman Sidney 
Published: November 2023

Abstract
The endoribonuclease RNase Pis a ubiquitous and essential ribonucleoprotein. It cleaves the 5’ terminal leader sequences of precursor tRNAs (ptRNAs) to generate mature tRNAs (Figure 1). Much of our current understanding of this enzyme is derived from studies of RNase P from Escherichia coli and Bacillus subtilis (1, 2). In E. coli, RNase P consists of an RNA subunit (Ml RNA, 377 nucleotides (nt)) and a protein subunit (CS protein, 119 amino acid residues). Ml RNA and CS protein are encoded by the rnpB and rnpA genes, respectively. Apart from ptRNAs, the precursors to 4.5S RNA (p4.5S RNA) (Figure 1) and lOSa RNA (H. Inokuchi, personal communication) are also processed by RNase P from E. coli in vitro and in vivo. The observation that Ml RNA can catalyse the hydrolysis of ptRNAs in vitro even in the absence of CS protein demonstrated that RNase P owed its catalytic potential to its RNA subunit and that the protein subunit is a co-factor (3). How ever, both Ml RNA and CS protein are essential for the activity of RNase P in vivo. This latter observation emphasizes the importance of auxiliary role(s) played by the protein subunit in vivo (4, 5).
Study of RNA-protein recognition by in vitro selection
Authors: Bartel David P , Szostak Jack W 
Published: November 2023

Abstract
A major focus of biochemistry and genetics has been to deduce the determinants of molecular recognition and catalysis. In vitro selection of nucleic acids is a method that represents a synthesis of these two disciplines - a method that retains both the control of in vitro biochemical manipulation and the power of genetic selection. This new tool has been used successfully in many laboratories for applications ranging from defining DNA-binding sites of transcription factors to isolating new ribozymes. The basic features and applications of in vitro selection of nucleic acids have been reviewed extensively (1-6). After a brief overview of these features and applications, this chapter focuses on practical considerations for designing, implementing, and interpreting in vitro selection schemes for the characterization of RNA-protein interactions.
PFGE analysis of yeast artificial chromosomes
Authors: Ragoussis J 
Published: November 2023

Abstract
There are two basic pulsed field gel electrophoresis (PFGE)-related techniques for mapping yeast artificial chromosomes (YACs). One is to digest the DNA with restriction endonucleases, detect specific fragments with defined probes, and construct a restriction map. The other technique is to generate YAC variants—(so called fragmentation products)—by homologous recombination with vectors containing a repeated (e.g. Alu), or specific, sequence. Using the recombination potential of yeast allows the construction of YAC derivatives for mapping of sequences within a YAC.
Analysis of the genomes of protozoan parasites using PFGE
Authors: Kemp David J , Cappai Roberto 
Published: November 2023

Abstract
Pulsed field gel electrophoresis (PFGE) has played a major role in developing our current knowledge of the genomes of a number of protozoan parasites as the genomes are small and the chromosomes usually fall within a usable size range. The studies on malaria and trypanosomatids described here demonstrate the applicability of the techniques to other protozoa.
Desalting inositol polyphosphates by dialysis
Authors: van der Kaay Jeroen , van Haastert Peter J M 
Published: November 2023

Abstract
In the study of the large spectrum of inositol phosphates and the enzymes involved in their metabolism, anion-exchange chromatography is commonly used for the separation, identification and quantitation of inositol phosphates. Desalting inositol phosphates after chromatography is often desired. The most commonly used method for desalting involves chromatography using a small column packed with mixed ion-exchange resin from which the inositol phosphates are washed with a volatile eluent (e.g. ammonium formate, HCI or Freon/octylamine) which is subsequently removed by vacuum concentration. Other methods used for desalting are gel-filtration or HPLC using a volatile eluent. The large volumes which have to be dried down make this procedure time-consuming.
Applications of chemically synthesized RNA
Authors: Naryshkin Nikolai A , Smith terence v. , Gait Michael J , Earnshaw David J , Farrow Mark A , Fogg Jan H , Grenfell Richard L 
Published: November 2023

Abstract
Many structural and biochemical projects on RNA and RNA:protein interactions have benefited from recent significant improvements in methods of chemical and enzymatic synthesis of oligoribonucleotides(1-6). These methods can also now be used to introduce site-specifically into RNA a range of nucleotide analogues containing modified functional groups or attachment sites for cross-linking (7-10). lt is not unusual in a new project involving RNA for all the three oligoribonucleotide synthesis techniques described in this book to be used together, namely chemical synthesis (this chapter), in vitro transcription (Chapter 2), and ligation (Chapter 3).
The electrophoretic mobility shift assay for RNA binding proteins
Authors: Black Douglas L , Chan Raymond , Min Hosung , Wang Jiwu , Bell Leslie 
Published: November 2023

Abstract
This chapter will discuss the identification and characterization of RNA binding proteins using the electrophoretic mobility shift assay (EMSA). The basis of EMSA is the change in the electrophoretic mobility of a nucleic acid molecule upon binding to a protein or other molecule. Initially a labelled RNA, containing the binding sequence of interest, is incubated with a sample containing RNA binding proteins and then analysed on an electrophorctic gel. The unbound RNA will have a characteristic electrophoretic mobility on a non-denaturing or native gel. If the RNA is bound by one or more proteins or other molecules, the movement of the RNA through the gel is retarded. This gives rise to a characteristic shift in the position of the RNA hand on the gel. The mobility shift method is a common technique for identifying DNA binding proteins (1).
Assays for virus infection
Authors: Gray J J 
Published: November 2023

Abstract
The laboratory diagnosis of virus infections utilizes a wide range of techniques, including virus isolation (Chapter 2), the detection of virus antigens and specific antiviral antibodies and the amplification and detection of virus genomes. This chapter will outline the principal methods used for the diagnosis of virus infections, and describe in detail those which are used most commonly. This section will outline methods used to identify and quantify viruses grown in cell culture, since detailed methods for the isolation of viruses are described in Chapter 2. Many of the cytopathic effects (CPE) seen in virus-infected cell cultures are characteristic of specific viruses or virus groups (Table 1), and the ability to recognize a specific CPE is the important first step in virus identification. Not all viruses growing in cell culture produce a CPE, and other methods have been devised for their identification (Section 2.7). The object of these assays is to measure the number of infectious particles in a suspension of viruses. This is done either by plaguing viruses on monolayer cell cultures in order to reveal and enumerate plaques which originate from a single infectious virus particle (pfu) (Protocol I), or by determining the highest dilution of the virus suspension which produces a CPE in 50% of the cell cultures inoculated (TCID50) (Protocol 2). The TCID50 value is calculated by use of the Reed-Muench formula or the Karber method (1). Calculation of the TCID50 is more commonly used to determine the dose of virus required in a neutralization test, although plaque assays give a more precise measurement of virus infectivity.
Virus vaccines
Authors: Jennings R , Potter C W 
Published: November 2023

Abstract
Any list of significant developments in the history of medicine would include the introduction of vaccines against infectious disease. History records epidemics and pandemics in earlier centuries which ravaged society and influenced or precipitated major events. The eighteenth century saw the dawning of the age of immunization, the nineteenth the wider introduction of vaccines against a variety of human and animal infections, through the work of many microbiologists. The present time has seen the eradication of smallpox, the predicted eradication of poliomyelitis and measles, and the control of many other infectious diseases. Vaccine development has been targeted to all virus infections which have a significant impact on human and animal health in terms of morbidity and mortality. The vaccine era is considered to have begun with the use of vaccinia virus to immunize against smallpox in the eighteenth century, and continued and expanded with the development of inactivated vaccines and live attenuated vaccines. Novel, modern vaccination strategies of the late-twentieth century are based on genetic engineering and the application of molecular biology, including the development of recombinant virus vaccines, replication defective and DNA vaccines, together with the use of natural or fabricated accessory molecules and formulations for enhancing and directing the immune response. Table 1 lists many of those virus vaccines presently commercially available for the immunization of man and animals, their form or type, and also indicates some virus vaccines currently under development and which may be generally available in the future.
Development of RNA virus vectors for gene delivery
Authors: Usmani B A , Fassati A , Dickson G 
Published: November 2023

Abstract
Experimental gene transfer and clinical gene therapy approaches rely on the efficient delivery of genes to desired target cells. The first approved human gene transfer study began almost a decade ago using an RNA virus vector derived from a murine oncoretrovirus to deliver a bacterial marker gene to patients’ cells (1). This was rapidly followed by protocols to treat severe combined immunodeficiency disease (SCID) and advanced melanoma. There are now hundreds of human gene therapy trials ongoing to treat many different classes of illness including inherited genetic disorders, cancer, virus infections, and cardiovascular disease, the majority of which use modified retrovirus vectors as gene delivery vehicles.
G proteins and their identification
Authors: Mullaney Ian 
Published: November 2023

Abstract
The demonstration that G proteins mediate many intracellular signalling processes has led to the development of specific techniques that can be used to identify which of these polypeptides is involved upon receptor activation by ligand. This chapter deals with the methodology involved to produce specific immunological tools, and the use of these reagents to probe functionally the specificity of receptor-G protein interaction.
Adenylyl cyclases and cyclic AMP
Authors: Ho Maurice K C , Wong Yung H 
Published: November 2023

Abstract
Adenylyl cyclase (AC; adenylate cyclase or ATP pyrophosphate lyase; E.C. 4.6.1.1) catalyses the formation of cyclic AMP (cAMP), using ATP as substrate. The theoretical topology of all ACs is similar, with both amino- and carboxyl-termini on the cytosolic side, twelve transmembrane domains divided into two groups, and linked with an extended cytosolic loop (1). The catalytic activity and the regulatory elements of AC appear to be concentrated on the cytosolic loop (C1 domain) and the carboxyl-terminal tail (C domain). To date, nine subtypes of mammalian adenylyl cyclases (A Cs) have been successfully cloned (1-15), and a partial cDNA fragment corresponding to a novel AC subtype (type X) has also been identified (16). All ACs are subjected to extensive regulatory signals, including Ca2+, G proteins, protein phosphorylation, and small molecules such as forskolin and adenosine analogues (also known as P-site analogues), as reviewed in refs. 17-19, and summarized in Table 1.
Phosphoinositide 3-kinases
Authors: Anderson K E , Stephens, L R , Hawkins P T 
Published: November 2023

Abstract
Phosphoinositide 3OH-kinases (PI3Ks) are enzymes which can phosphorylate one or more membrane inositol lipids in the 3-position of the inositol ring; in vitro they can make phosphoinositide(3)phosphate (PtdIns(3)P), phospho- inositide(3,4)bisphosphate (PtdIns(3,4)P2), and phosphoinositide(3,4,5)triphos- phate (PtdIns(3,4,5)P3) from phosphoinositide (Ptdlns), phosphoinositide(4) phosphate (PtdIns(4)P), and phosphoinositide(4,5)bisphosphate (Ptdlns(4,5) P2), respectively. The substrate specificity of PI3Ks in the intact cell is a matter of current debate, but most evidence suggests that the preferred substrate of the sub-family of PI3Ks that are agonist sensitive (type 1 PI3Ks) is Ptdlns (4,5)P2, and also possibly PtdIns(4)P. Hence the rapid rise in the levels of PtdIns(3,4,5)P3 and PtdIns(3,4)P2 seen on stimulation is widely regarded as the major signal generated by this pathway.
Signal transduction by sphingosine kinase
Authors: Heringdorf Dagmar Meyer Zu , Koppen Chris J Van , Jakobs Karl H 
Published: November 2023

Abstract
Several new findings in the last few years have drastically challenged our view of the role of sphingolipids in cellular function. It is now evident that not only the complex sphingolipids of the plasma membrane, but also small molecules of sphingolipid metabolism, play an essential role in cellular regulation. Analogous to the breakdown products of glycerophospholipids, certain small sphingolipids are messenger molecules which transduce information both between cells and within cells. Of these, ceramide and sphingosine-1- phosphate (SPP) have gained major attention (1), although others may also have a signalling function. Ceramide is cleaved from sphingomyelin by sphingomyelinase(s) (see Figure 1), and apparently plays an important role in apoptosis. SPP was originally considered as an intermediate product in the breakdown of sphingosine, but now it is recognized that it controls a variety of cellular functions.