bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lipoprotein turnover and metabolism
Authors: Gaw Allan , Packard Christopher J , Shepherd James 
Published: November 2023

Abstract
The study of lipoprotein kinetics has added significantly to our understanding of the regulation of the plasma lipid transport system and the aberrations that give rise to hyperlipidaemia. Lipoprotein metabolism can be divided into three inter-related pathways (Figure 1). The first, associated with the synthesis and catabolism of chylomicrons, is responsible for the absorption of dietary fat and delivery of triglyceride to peripheral tissues (such as adipose tissue and skeletal muscle) and cholesterol to the liver. The second involves the production of very low density lipoproteins (VLDL) from liver and their remodelling to intermediate and low density lipoproteins (IDL, LDL). By this pathway triglyceride and cholesterol are transported in the fasting state. The third is concerned primarily with the return of cholesterol from peripheral sites to the liver and is mediated by the high density lipoproteins (HDL). The kinetics of these processes have been followed using both lipid and apolipoprotein markers. The former were used in early experiments and provided information on total body turnover rates of cholesterol and triglyceride. The latter have yielded data on the behaviour of individual lipoprotein fractions. Chylomicrons, VLDL, IDL, and LDL contain apolipo protein B as their major structural protein. This high molecular weight polypeptide exhibits an extended secondary structure that intercalates into the lipoprotein surface and remains there throughout the lifetime of the particle in the plasma. Hence it acts as a marker of a particle ‘s metabolic fate, whereas all other lipoprotein constituents including the A, C, and E apolipoproteins exchange freely between particles, complicating inter pretation of their metabolic properties.
PCR with highly degenerate primers
Authors: Mcpherson Michael J , Jones Kerrie M , Gurr Sarah Jane 
Published: November 2023

Abstract
Primers for use in the polymerase chain reaction are usually unique oligo-nucleotides designed from a known DNA sequence. However, for an uncloned gene, where obviously no DNA sequence data are available, the PCR may be used in cloning strategies by designing highly degenerate primers from protein sequence data (1–3). Such degenerated primers have been used successfully to isolate members of multigene families and genes from diverse species. The amplified DNA represents part of the gene of interest and can therefore be used as a homologous hybridization probe, allowing high stringency screening of gene libraries to isolate corresponding cDNA or genomic clones. The PCR amplified DNA may also be used to probe Southern or Northern blots.
The fidelity of DNA polymerases used in the polymerase chain reactions
Authors: Eckert Kristin A , Kunkel Thomas A 
Published: November 2023

Abstract
Purified DNA polymerases are one of the primary tools for molecular biology. Yet choosing the most appropriate DNA polymerase for any particular application requires an understanding of the substantial biochemical differences between the available enzymes. One aspect of interest for PCR is the fidelity of DNA polymerization, i.e. the number of errors produced per nucleotide synthesized. We begin this chapter by discussing the enzymology of error discrimination by DNA polymerases during DNA synthesis in vitro. We then describe the fidelity of several prokaryotic DNA polymerases using a simple assay for scoring DNA polymerase errors. Next we examine the parameters that affect the fidelity of the polymerase most often used for PCR, the Taq DNA polymerase isolated from Thermus aquaticus.
Intracellular trafficking in fission yeast
Authors: Armstrong John , Fawell Erica , Pidoux Alison 
Published: November 2023

Abstract
The analysis of protein targeting and membrane traffic in eukaryotic cells has benefited greatly from the use of genetic methods in the budding yeast Saccharomyces cerevisiae. The approach was introduced by Novick and Schekman, whose sec mutants define a collection of genes which function at different stages of the secretory pathway. These gene products are now the object of intense study (reviewed in reference 1); already it is clear that the general characteristics of membrane traffic are conserved between yeast and higher eukaryotes, and that several of the sec genes have homologues in mammalian cells. Genetic screens in yeast have since been used to identify mutants in other processes of membrane traffic, such as protein targeting to the vacuole (2, 3) and retention of proteins in the endoplasmic reticulum (4). Given the accumulation of biochemical and genetic tools now available, it is likely that S. cerevisiae will continue to be popular for studies of different aspects of protein targeting. Several excellent collections of protocols for genetics, cell biology, and biochemistry in budding yeast are now available (e.g. 5, 6).
In vitro reconstitution of endocytic vesicle fusion
Authors: Gruenberg Jean , Gorvel Jean-Pierre 
Published: November 2023

Abstract
It is now well established that, after internalization by clathrin-coated pits, solutes and receptors first appear in early endosomes at the cell periphery, then in late endosomes in the perinuclear region, and eventually in lysosomes (reviews in references 1-3). We have observed that tracers leaving the early endosomes appear in large spherical vesicles before they reach late endosomes (4). These large vesicles resemble one of the subpopulations of multivesicular endosomes described by Dunn et al. (5). Tracers internalized after microtubule depolymerization pass through early endosomes and reach these large vesicles but they do not appear in the late endosomes. We thus proposed that these vesicles are involved in the microtubule-dependent transport between early and late endosomes and may correspond to the vesicles observed to move in vivo between the cell periphery and the perinuclear region. In this chapter we will refer to these vesicles as endosomal carrier vesicles.
The extracellular matrix—an overview
Authors: Haralson Michael A , Hassell John R 
Published: November 2023

Abstract
The extracellular matrix (ECM) has been traditionally thought of as the structurally stable material that provides support for cells and tissues. However, a number of discoveries over the past two decades have changed this perspective and established the importance of the ECM in modern biology. First, both biochemical and molecular biological investigations have documented that four major classes of macromolecules—the collagens, proteoglycans, structural glycoproteins, and elastin—collectively comprise the ECM of animal cells (1—6). Furthermore, with the exception of elastin, each class of matrix macromolecules has been found to contain families of related proteins with each member being a unique gene product. Second, individual members of each class and family of ECM molecules were found to exhibit a degree of tissue-specific distribution implicating the matrix in development and tissue function (7—9). Third, specific cell surface receptors for ECM components were identified, which provided a rational basis for linking the ECM with the biology of the cell (10—16). From these discoveries we now appreciate that the extracellular matrix is a complex ordered aggregate composed of a number of different macromolecules whose structural integrity and functional composition are important in maintaining normal tissue architecture, in development, and in tissue specific function (2, 4, 6, 8, 17). Finally, in conjunction with these studies has come the recognition of the importance that dysfunctional matrix components and abnormalities in ECM biosynthesis and catabolism have in both inherited and acquired diseases and in normal wound healing (2, 3, 5, 6, 9, 18).
Elastin
Authors: Davidson Jeffrey M , Giro Maria Gabriella , Mecham Robert P 
Published: November 2023

Abstract
Elastin, when mature, is the most insoluble protein in the vertebrate body. It is synthesized as an Mr= 70 000 precursor from a 3.5 kb mRNA encoded by a single gene into a protein termed tropoelastin, which is cross-linked sub-sequent to secretion by the copper-dependent enzyme, Iysyl oxidase (1). Therefore, for most purposes elastin purification simply requires the solubil ization and removal of other protein components from connective tissue to leave elastin as the remaining residue. The elastic.fibre, however, consists of insoluble elastin plus other structural macromolecules, collectively referred to as microfibrillar components (2). Since these molecules are also relatively insoluble, their isolation is also accomplished by selective extraction of the elastic fibre under denaturing and reducing conditions. This chapter will not detail the isolation of these other components.
Use of extracellular matrix and its components in culture
Authors: Kleinman Hynda K , Kibbey Maura C , Cannon Frances B , Weeks Benjamin S , Grant Derricks 
Published: November 2023

Abstract
The interaction of cells with the extracellular matrix is an important determinant of cell behaviour (1). Extracellular matrices not only provide structural support but also elicit biological responses which are mediated by specific components of the matrix through interactions with cell surface receptors. Matrices contain collagens, proteoglycans, and glycoproteins. The type and amount of each component depend on the cell type with which it is associated. Our understanding of the biological activity of the extracellular matrix and its components has been enhanced by the ability to purify the components and to examine their effects on cells in culture. From these studies, it is clear that components such as collagen types I and IV, laminin, and fibronectin influence cell behaviour in a cell specific manner. Cells adhere, grow, migrate, and differentiate on matrix components, and, in many cases, do so better than on plastic or glass surfaces. Specific sites on matrix molecules are being defined for these processes with distinct domains, and synthetic peptides having been shown to possess biological activity (2). Specific cellular receptors for each matrix macromolecule have also been defined. This chapter focuses on the use of basement membrane preparations and the components laminin and collagen in cell culture (3, 4). Many of the techniques described herein can be directly adapted to the other matrix glycoproteins including fibronectin, entactin, vitronectin, thrombospondin, tenascin, osteopontin, and osteonectin.
Isolation and analysis of RNA from connective tissues and cells
Authors: Sandell Linda J 
Published: November 2023

Abstract
Extraction of intact functional RNA from connective tissues presents special challenges. Most extracellular matrix (ECM) molecules are very large and consequently their mRNAs are among the longest known. The difficulty in isolating these large mRNAs is compounded by the large amount of ECM surrounding the cells. In addition, the number of ECM mRNAs per cell may be quite low due to the low turnover rate of the protein. In some connective tissues, however, where the ratio of cells to matrix is high, RNA can be isolated from the tissue directly. In other cases where the matrix is resistant to denaturation or where the matrix is particularly abundant, better results are achieved when the matrix is removed from the cells before isolation of RNA. In this chapter methods will be described that have been used successfully to isolate RNA from cartilage and a variety of other connective tissues, such as bone, tendon, and smooth muscle, and from connective tissue cells in culture.
Generation of cosmid DNA libraries and screening for extracellular matrix genes
Authors: Toman David , Crombrugghe Benoit De 
Published: November 2023

Abstract
The application of molecular biological techniques to study the structure, function, and transcriptional regulation of extracellular matrix genes has provided a wealth of knowledge at the DNA level. A nine base pair repeat encoding Gly-X-Y at the amino acid level has been implicated as the ancestor of the collagen molecule. This repeat has been assembled into a 54 base pair exon which has been duplicated and combined with other functional groups to generate at least part of the family of collagen genes. Individual members of the collagen family show tissue specificity and provide different structural roles in many multicellular organisms. Analyses of the role of collagens in human connective tissue diseases have shown a wide spectrum of phenotypic effects which are currently studied in order to more fully understand their functional role in an organism. Mouse transgenic models are probing structure/function relationships with other components of the extracellular matrix. Identification and characterization of specific transcription factors and cis-acting elements are unravelling the regulatory pathways which determine the tissue- and developmental-regulation of expression of these genes.
Stable Isotope Labelling and Resonance Assignments In Larger Proteins
Authors: Markley John L , Kainosho Masatsune 
Published: November 2023

Abstract
This chapter introduces the general principles of isotope-assisted NMR spectroscopy of proteins and presents descriptions of several specific procedures. The rationale and methodology of protein labelling are discussed, and NMR experiments that exploit various patterns of labelling are surveyed.
Insulin synthesis
Authors: Yes Elaine M Bail , Guest, Paul C , Hutton John C 
Published: November 2023

Abstract
The structure of insulin has been highly conserved throughout higher vertebrate evolution. The hormone consists of an A-chain (21 amino acids) and a B-chain (30 amino acids), connected by two intermolecular disulphide bonds (A7-B7, A20-Bl9), with an intramolecular bond between A6 and All. Invariant features include the positions of the three disulphide bonds, the N- and C-terminal regions of the A-chain, and the hydrophobic residues in the C-terminal region of the B-chain. In the presence of zinc, at acid pH, most insulins form crystals composed of insulin hexamers. The hexamer consists of three dimers arranged roughly in a plane. The central axis perpendicular to the plane passes through two zinc ions; each is co-ordinated with the imidazole groups of three BIO histidine residues with one located above and one below the plane of the hexamer. The insulin dimers are held together in the crystal by hydrogen bonds between amino acid B24 and amino acid B26, forming an antiparallel 13-pleated sheet structure (Steiner et al. 1989).
The insulin receptor
Authors: Siddle Kenneth 
Published: November 2023

Abstract
It has been widely accepted for many years that the metabolic effects of insulin are initiated by the binding of insulin to cell-surface receptors. It has become clear that there is only one type of insulin receptor, encoded by a single gene. The structure and activity of this receptor is understood in considerable detail, although many important aspects remain to be eludicated, from the mechanism of receptor activation at the molecular level to its involvement in pathophysiological processes in vivo.
Introduction to diabetes
Authors: Turner Robert , Neil Andrew 
Published: November 2023

Abstract
Diabetes is the name given to the clinical description of patients with a number of symptoms arising from raised glucose levels. The syndrome can occur from many specific ‘secondary causes’, including pancreatectomy, iron overload of the !3-cells resulting from haemochromatosis, excess cortisol production in Cushing’s syndrome, and excess growth hormone secretion in acromegaly and insulin-resistant syndromes, including insulin-receptor defects. The majority of patients do not, however, have any of these identified defects and are said to have ‘idiopathic’ diabetes. The metabolism of glucose involves many organs, in each of which several important metabolic steps occur. Key regulatory points include hepatic regulation of glucose uptake and release, muscle utilization of fuels, control by pancreatic production of insulin and glucagon, and neurogenic controls. Diabetes may arise from abnormalities at one or several sites in the complex feedback loops in this system. Two main types of diabetes can be distinguished. Different nomenclatures for these types have arisen because overlap between the types makes a strict, simple classification covering all patients impossible.
Aetiology of type II diabetes
Authors: Cerasi Erol 
Published: November 2023

Abstract
Type II diabetes is certainly the most common form of the disease, even in Scandinavia where the prevalence of type I diabetes is remarkably high. The past decades have seen an increase in type II diabetes in Western societies, which cannot be accounted for solely by improved diagnostic procedures or increased longevity (Everhart et al. 1985). In addition, the traditionally low prevalence of type II diabetes in the Far East and in developing countries is rapidly becoming a feature of the past, due to the ongoing improvement in standards of living. It seems certain, therefore, that humanity will enter the twenty-first century with at least twice as many type II diabetics as today, unless curative measures can be established during this decade.
Development, technology, and utilization of capillary columns for gas chromatography
Authors: Dawes Peter A 
Published: November 2023

Abstract
The concept of using hollow tubes as chromatography separation columns stemmed from a theoretical mathematical investigation and later experimental work by Marcel Golay in 1957. The more correct terminology for this invention is open tubular capillary column but the shortened and more general terminology of ‘capillary column’ has found its way into common use. The most obvious benefit of capillary columns over packed columns was the increased resolving capacity. An examination of the van Deemter equation will yield several reasons why a capillary column can have higher resolving power than a packed column. A large contributing factor is, however, simply the increased length.
Gas chromatography in analytical toxicology: principles and practice
Authors: Flanagan Robert j. 
Published: November 2023

Abstract
Analytical toxicology is the detection, identification and, if appropriate, measurement of drugs and other poisons in biological and other relevant specimens to aid the diagnosis, treatment, prognosis, and in some cases the prevention of poisoning. The specimens encountered may range from the relatively simple, such as expired air, to amongst the most complex of samples, the residue from a decomposed body. This being said, most work is performed on commonly available samples such as blood and urine. In analytical toxicology, gas chromatography (GC) has three principal advantages over other widely-used techniques such as high-performance liquid chromatography (HPLC) and immunoassay. First, GC has a range of sensitive detectors [‘universal’: flame ionization detector (FID); selective: nitrogen/phosphorus detector (NPD), electron capture detector (ECD)] which can be used in parallel. Second, high efficiency (capillary) GC columns are now widely available. Third, GC is easy to interface with techniques giving direct information about compound identity such as mass spectrometry (MS) and Fourier-transform infrared spectrometry (FTIR). With GC, as with HPLC, qualitative and quantitative information can often be obtained simultaneously.
Gene promoter analysis of malting enzymes
Authors: Skriver K , Olsen F L , Leah R , Mundy J 
Published: November 2023

Abstract
Malt hydrolase production is primarily determined by the rate of hydrolase gene expression, which is controlled by the antagonistic hormones gibberellin (GA) and abscicic acid (ABA). We have studied this mechanism in aleurone cells transformed with gene constructs which are responsive to GA (a-amylase), ABA (rab), or which are insensitive to the hormones (chitinase & lipid transfer protein). We have thereby defined gene regulatory regions determining hormone sensitivity, and those which confer constitutive, high level expression in aleurone cells. These DNA elements may be used to design malting varieties with novel patterns of hydrolase production by genetic engineering.
Non conventional methods for rapid production and malting/brewing quality evaluation of new barley varieties
Authors: Orive M , Batalla G , Cerveró J A , Torrent J 
Published: November 2023

Abstract
Increased efficiency in barley breeding programs would offer maltsters and brewers a wider, high quality range of brewing varieties. Thus ANTHER CULTURE is optimized for routine application in commercial breeding providing pure line field material 1^ years after crossing. For quality screening of this scarce seed, rapid and highly discriminative barley/malt analyses (NIR, HPLC, SMALL SCALE METHODS) are developed and validated. RFLP’s, ISOZYMES and HORDEINS would allow selection 1 year after crossing. Therefore isozyme and protein capillary electrophoretic, HPLC patterns are obtained. Since they maximize discriminatory power of hordein electrophoresis, rapid, objective identification of individual varieties is achieved.
Advances in the fermentation of high gravity wort
Authors: D’Amore Tony , Celotto Guy , Stewart Graham G 
Published: November 2023

Abstract
The fermentation of high gravity wort (16°P to 34°P) by a lager yeast strain has been investigated. As the wort gravity is increased, stress effects such as high osmotic pressure and elevated ethanol levels become more apparent, resulting in reduced yeast viability and incomplete fermentation. The fermentation of 25°P wort on a pilot plant scale (30 1 to 16 hl) was successfully performed by altering such parameters as the fermentation temperature, wort oxygen level and yeast pitching rate. Differences in the production of acetaldehyde, esters, higher alcohols and intracellular trehalose were observed. The resulting beers were found to be comparable to commercial products.
Present and future legislation of food hydrocolloids
Authors: Gray P S , Penning W 
Published: November 2023

Abstract
The development of the EC legislative system and programme of food law harmonisation is described and the roles of the various institutions the Commission, the Council, the Parliament and the Court of Justice is explained. The numerous judgements of the Court of Justice on food are briefly dealt with giving special attention to their effect on national food law and the way in which the are complementary to the harmonisation programme.
Gel profile star diagrams as a technique for the visualisation of gel texture
Authors: Sworn Graham 
Published: November 2023

Abstract
Nine key characteristics of alginate gels have been measured using Texture Profile Analysis. These are; Modulus, Hardness and Brittleness at acid and neutral pH, Hardness with high and low methoxyl pectin, and Gel Strength Build-up. Syneresis has also been measured. From this data 8 point Gel Profile Star Diagrams have been constructed which illustrate the effects of mannuronic/guluronic acid ratio, degree of calcium conversion, pH, and type of pectin.