bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cell synchronization as a basis for investigating control of proliferation in mammalian cells
Authors: Stein Gary S , Stein Janet L , Lian Jane B , Last T J , Owen Thomas , Mccabe Laura 
Published: November 2023

Abstract
From a historical perspective, synchronized cells have provided the basis for defining regulatory mechanisms operative during specific periods of the cell cycle; for example, those at key transition points that separate Gi, S-phase, G2 and mitosis as well as those operative when quiescent cells are induced to proliferate. Cell synchrony studies have also contributed to establishing modifications in gene expression which are functionally related to initiation and execution of DNA replication during S-phase and to the onset and completion of mitotic division. More recently this experimental approach has increased our understanding of key modifications in gene expression and the interrelated cellular signalling pathways that transduce, integrate and amplify information that support each stage of the cascade of regulatory events requisite for progression through the cell cycle. This chapter is restricted to methods that have proven to be effective for synchronizing continuously dividing normal diploid and tumour cells as well as for induction of quiescent cells to embark upon a progression of events that leads to cell division. Protocols that can be applied for monitoring cell synchrony will be addressed. In addition, we will evaluate the extent to which cell synchrony can be achieved in a manner that is compatible with physiological regulation that is operative under in vivo conditions.
Analysis of the effects of purified growth factors on normal human fibroblasts
Authors: Phillips Paul D , Cristofalo Vincent J 
Published: November 2023

Abstract
Normal human diploid fibroblast-like cells are widely used for studies of in vitro cell ageing and in studies of growth factor action and cell proliferation. Because of their special growth characteristics the manipulation of these cells requires special attention and understanding to ensure the vigorous and optimum growth of the cultures.
Cells from normal human tissues, explanted and grown in tissue culture, have the following characteristics: (i) the cultures possess a finite and predictable proliferative capacity most accurately measured as cumulative number of population doublings (chapter 12); (ii) the replicative life span of a cell line is inversely proportional to the age of the donor; (iii) normal human fibroblast-like cells have never been shown to transform spontaneously in culture, i.e. they have never been shown spontaneously to acquire the indefinite proliferative capacity characteristic of tumour derived or otherwise transformed cells; and (iv) normal human cells can be transformed into cultures with an indefinite proliferative capacity by infection with DNA tumour viruses such as simian virus 40 (SV40) (1, 2, and Chapter 12).
Epithelial cell culture
Authors: Shaw Andrew J 
Published: November 2023

Abstract
Our understanding of the functions of epithelia, and their development and maintenance, has been rapidly advanced by the use of cell cultures which faithfully display the morphological, biochemical, and functional characteristics of these tissues. This book gives detailed step-by-step descriptions of the methods used to mimic epithelial and endothelial tissue iin vitror, and the techniques used to characterize these cell culture models, together with hints and tips for success, troubleshooting guides, and key background information. Topics covered include: modelling epithelial tissues in vitro; identification and localization of cytokeratins; assessment of cell polarity; xenobiotic metabolism; culture and characterization of human endothelial cells; studying transport processes in absorptive epithelia; a model of glandular epithelium for studying secretion; a model of the blood-testis barrier for studying testicular toxicity; reconstruction of human skin epidermis in vitro. Epithelial Cell Culture: A Practical Approach is a timely and comprehensive practical guide for all researchers who are studying, or intend to study, epithelia-related tissues using modern cell culture techniques. As such, this book will have a very wide appeal to cell biologists, developmental biologists, biochemists, pharmaceutical scientists, pharmacologists, and toxicologists.
Assessment of cell polarity
Authors: Hughson Elaine J , Hirt Robert P 
Published: November 2023

Abstract
The epithelial sheets of metazoa, such as those lining the cavities of the lung, the kidney, or the digestive tract, can be compared to the plasma membrane of unicellular organisms (1). They are the site of complex exchanges between the outside world and the internal milieu which are essential to maintaining a constant body fluid composition. Epithelial cells, therefore, in addition to forming a simple barrier, are the site of numerous vectorial transport processes which are the bases of functions such as water and ion homeostasis, metabolite uptake and secretion (2, 3), and ultimately, protection against pathogens (4).
Transcription on chromatin templates
Authors: Luse Donal , Felsenfeld Gary 
Published: November 2023

Abstract
As it has become clear that the coding regions of transcriptionally active chromatin are packaged in nucleosomes (1, 2, 3), it has also become important to determine how such a histone-covered template can accommodate to the passage of RNA polymerase. For this purpose, it has proved useful to employ ‘model’ systems, in which templates reconstituted from specific DNA sequences and histone octamers are transcribed by either prokaryotic or eukaryotic polymerases. Experiments with prokaryotic enzymes such as the T7 or SP6 polymerases give us information about the intrinsic behaviour of histone octamers during transcription; they obviously cannot reveal possible contributions to the reaction that might be made by a eukaryotic polymerase, or by specific factors capable of modifying nucleosome stability that might be present in eukaryotic cells. Experiments with eukaryotic polymerases have the limitation that in using purified enzymes these specific factors may have been excluded. Despite these limitations, much valuable information about the properties of chromatin templates has been obtained with both kinds of polymerases.
Epigenetic regulation in mammals
Authors: Tilghman Shirley M , Willard Huntington F 
Published: November 2023

Abstract
The term ‘epigenetic’ when applied to mammalian genes refers to different stable states of phenotypic expression usually thought to be due to differential effects of chromosome or chromatin packaging, and determined or influenced by control mechanisms which can be manifested over exceptionally long distances. Both the recognition of epigenetic states and the design of tractable experimental approaches to analyse them require an appreciation for mammalian gene expression in its dynamic setting. One must consider those aspects of chromatin assembly, structure, and replication (see Chapters 1-3) that determine whether, when, and at what level genes are expressed, the series of dynamic events that occur at each cell division, when and under what circumstances epigenetic states are stably transmitted through meiosis or mitosis, and when and under what circumstances epigenetic states are reversed in the course of normal development or as a result of mutation or chromosome rearrangement.
Isolation and growth of hyperthermophiles
Authors: Sharp Rirhard J , Raven Neild. H 
Published: November 2023

Abstract
This chapter discusses the problems encountered in the isolation and cultivation of hyperthermophilic microorganisms and presents procedures and guidance for working with the more common hyperthermophilic. We define hyperthermophilcs as organisms with growth optima of&gt; 75°C. This excludes thermophilic Bacillus and Thcrmus species which have recently been discussed in other reviews (1, 2). The chapter will describe procedures which have been applied to the growth of both hypcrthcrmophilic Archaea and Bacteria.
Fermenter Vent-Gas Analysis
Authors: Salmon Peter M , Buckland Barry C 
Published: November 2023

Abstract
Almost any microbial process involves the consumption or evolution of gases, and vent-gas analysis therefore provides a convenient means to monitor microbial activity. In contrast to other monitoring techniques for which the culture medium must be placed in direct contact with a sampling device or probe, vent-gas analysis poses no risk to sterile operation and has consequently become indispensable in both research and manufacturing (1).
Preparation of single cell suspensions
Authors: Cheetham K M , Shukla N , Fuller B J 
Published: November 2023

Abstract
The cell, as the basic functional unit within a tissue or organism, holds the potential for allowing us to probe the numerous interrelated processes which comprise overall function, or to manipulate specific pathways for scienlif1c or commercial gain. As such, the rapid expansion over the last two decades in cell applications in fields such as biochemistry, molecular biology, and biotechnol ogy has led to an equivalent effort in devising methods for cell isolation and handling. Although many important studies have been performed on continu ously growing cell lines (which may often he commercially available), many other investigations can only be performed on cells from selected intact tis sues, expressing particular differentiated functions. Thus the ability to liberate high yields of viable cells from the structural constraints of their native surroundings is often the first hurdle to overcome in experimental biology. Section 2 describes these methods of dispersal. These are then complemented by the need to ensure that the process has yielded individual cells which have retained metabolic and functional capabilities. Section 3 describes methods of viability assessment. Section 4 provides methods for separating viable and non-viable cells from the isolate, which invariably is a mixture of populations containing the desired cell type. Methods of cell characterization arc described in Section 5. In the current chapter, we have addressed these particular prob lems, giving a variety of examples from currently used techniques.
Applications of Antisense Technology in Plants
Authors: Kuipers Anja G J , Jacobsen, Evert , Visser Richard G F 
Published: November 2023

Abstract
In 1984, Izant and Weintraub (1) published the first experiments on the antisense effect of gene constructs in eukaryotic cells. Since then, the inhibition of gene expression via antisense RNA has developed into a technique that is widely applied in molecular biology. In the field of plant biotechnology and plant molecular biology Ecker and Davis (2) were the first to describe a transient assay system in which carrot protoplasts were simultaneously electro-porated with sense and antisense CAT genes. This resulted in the inhibition of CAT activity of up to 95% as compared to the control which only contained a sense CAT gene. These initial studies were followed by experiments with various plant species, in which the expression of a wide variety of genes has been effectively inhibited, either transiently with in vitro transcribed antisense RNAs, or by the introduction of antisense genes via transformation (reviewed in refs 3-5).
Antisense Molecules and Ribozymes: Medical Applications
Authors: Strauss Michael 
Published: November 2023

Abstract
The application of antisense oligodeoxynucleotides as tools for specific inhibition of gene expression was an extremely important step in the development of molecular medicine (reviewed in ref. 1, see also Chapters 3 and 11). From the first moment of experimental investigation of the power of these molecules it was obvious that, at some stage, this class of molecules would find its way to clinical application as therapeutic drugs (2). After several years of successful use in cell culture experiments, antisense molecules including synthetic oligos and expressed RNA are increasingly applied in animal experiments. Most of the studies carried out in cdl culture and animals so far have been aimed at the investigation of the consequences of the loss of a particular function. The antisense strategy has the advantage over the powerful method of gene knock-outs of allowing transient suppression of gene expression. Thus. it allows one to inactivate, for a short period of time, those functions which would be lethal if inactivated at the gene level (see also Chapter 12 for another approach to the problem of lethality). Since the power of the antisense approach first became obvious, studies focusing on the treatment of diseases have been initiated over the last few years. It seems only a matter of time before this strategy can be applied clinically to the treatment of particular diseases, e.g. infectious diseases like hepatitis B and C, or HIV infection, and even some forms of cancer. This chapter will give a brief overview of the different fields of medical application of antisense molecules, and will provide the reader with protocols for the experimental use of these molecules in cell culture and animal studies.
Measuring antibody affinity in solution
Authors: vadi-ohaniance Lisa dja , Goldberg michele. , friguet bertrand 
Published: November 2023

Abstract
Determination of the affinity of a monoclonal antibody (mAb) for its antigen (Ag) is of considerable importance. It gives a quantitative indication of how strong the interaction is between the Ab and the Ag. It is the basic experi mental parameter in a variety of studies, such as the analysis of mAb/Ag binding mechanism (1, 2) or the use of mAbs as conformational probes (3, 4). Hence the need for convenient and rigorous methods to determine the affin where Kd is the equilibrium dissociation constant, [complex] is the concentra tion of saturated antigen or mAb sites, [Ag] is the concentration of free anti genic sites on the antigen, and [Ab] is the concentration of free binding sites on the antibody. These concentrations are binding site concentrations in the solution at equilibrium. This has three important consequences, as described in Sections 1.1-1.3.
Isolation and fractionation of chromatin and linker histones
Authors: Thomas Jean O 
Published: November 2023

Abstract
Nuclei from different sources usually have a common pattern of core histones (components of the histone octamer) as revealed by SDS/polyacrylamide gel electrophoresis, but in some cases they have distinctive linker histone variants that may correlate with particular states of cell differentiation and transcriptional activity (more subtle Hl subtypes present in many cell types are not resolved in SDS gels). Nuclei from different sources may also differ in nucleosome repeat length. Procedures are described for the isolation of nuclei from rat liver, and from chicken erythrocytes and sea-urchin sperm which contain extreme HI variants and have repeat lengths longer than the canonical 200 base pairs of rat liver and many other tissues. A method is also described for the isolation of nuclear populations from various mammalian brain cell types; the distinctive feature here is that neurones from the cerebral cortex have a shorter repeat length than the glial cells from the same tissue or cerebellar neurones, and only half the Hl content.
Electron microscopy of chromatin
Authors: Thoma Fritz 
Published: November 2023

Abstract
Electron microscopy (EM) has been used to investigate how DNA is folded in nucleosomes, chromatin fibres. and higher-order structures. In contrast to biochemical and biophysical techniques, which produce averaged information of a whole population of structures, EM allows one to visualize and analyse individual molecules or chromatin complexes; averaged information must then come from the analysis of a significant number of complexes.
Cytokinesis
Authors: Goldberg Michael L , Gunsalus Kristin C , Karess Roger E , Chang Fred 
Published: November 2023

Abstract
Cytokinesis is a multi-step process that proceeds linearly with time according to the following general framework. It begins with a signal from the mitotic apparatus that must be transmitted to the cellular cortex at the cleavage site. Receipt of the signal at the cortex sets off a complex set of reactions that transduce the signal into changes in the actin cytoskeleton and lead to formation of an actomyosin-based contractile ring. Mechanical processes in the ring and its interaction with the cortex result in cytokinesis. The distinctions between these steps are probably largely artificial. It is not clear, for example, whether some of the molecules we describe as transducing a cytokinetic signal are in fact part of the signal itself. Even this seemingly innocuous (though admittedly simplified) view of cytokinesis is not free of controversy, and some kinds of cytokinesis almost certainly do not conform to this pattern. Many questions remain unanswered and are addressed in the following sections.
Prediction of barley which causes premature yeast flocculation
Authors: Nakamura Takashi , Chiba Kazuhiro , Asahara Yumiko , Tada1 Setsuzo 
Published: November 2023

Abstract
During fermentation of wort produced from certain malts, premature yeast flocculation occurs leaving fermentable sugars in solution. Premature yeast flocculation results in a low attenuation. We have developed a novel fermentation method to predict barley which causes premature yeast flocculation. A barley extract obtained by hot-water extraction using an enzyme mixture of BÂ¬ glucanase, protease, a-amylase and B-amylase was added to a synthetic medium. The premature yeast flocculation activity of this extract was evaluated by a fermentation test. Analysis could be carried out within four days and was applicable to either dormant or water sensitive barley.
Production of aromatic hop varieties starting from ‘‘in vitro”
Authors: Vanhame! S , Putman R , Timmermans P , Everaert E , Cooman L De , Beeumen J Van , Oord R Van Den , Debergh P 
Published: November 2023

Abstract
The objective was to increase the return of the aroma hop Saaz starting from in vitro cultures. Saaz hops were cultivated on a hop garden in our brewery, in cold greenhouses, hothouses and plastic tunnels. Fingerprinting of various crops showed that they are identical with the original variety; chemical and sensorial analysis of pilot brews showed less differences. The influence of nutritive ingredients, length of the day and temperature on flower growth were evaluated. The formation of bitter acids and essential oils were studied in function of the developmental stage of the hop cones and cultivation method.
Enzymes from the biosynthesis of hop α- and β-acids
Authors: Fung S Y , Zuurbier K W M , Paniego N B , Scheffer J J C , Verpoorte R 
Published: November 2023

Abstract
The hop enzyme catalysing the first step has been purified; it is a new enzyme closely related to the enzyme chaicone synthase from plant flavonoid biosynthesis. Work on hop enzymes of the later stages is still in progress. Knowledge from the study of hop acid biosynthesis could lead to interesting applications in hop breeding.
Differential behaviour of cis- and trans-iso-α-acids
Authors: Hughes P S , Menneer I D , Walters M T , Marinova G 
Published: November 2023

Abstract
Recent analytical developments have allowed the routine separation of iso-a-acids. By applying such methodology, it has become apparent that there are differences in the behaviour of c/s- and trans-iso-a-acids. Thus, trans-iso-a-acids are lost to a greater extent during beer production and on subsequent storage, and are also partitioned into beer foam to a greater extent than their cis- counterparts. Whilst it is difficult to explain these differences on hydrophobicity grounds, it is possible that there is a thermodynamic rationale. These differences in behaviour are of practical significance, particularly in terms of optimizing hop acid utilization, bitterness delivery and foam quality.
The effect of different cereal coloured malts on reducing power during the brewing process
Authors: Griffiths D J , Maule A P 
Published: November 2023

Abstract
This project was designed to investigate the relative reducing power of barley, wheat, rye and oat crystal malts, the relationship with degree of colour formation and the reducing power levels during the brewing process when these malts were used. A specially designed 3 kg capacity mini-roaster was used to produce the crystal malts. Reducing power was measured using the Chapon rapid determination method (iron II dipyridyl complex) on a Tannometer. Indications are that measurement of reducing power during the brewing process will provide useful data towards maximising beer stability by selection of raw materials.
Modernization, rationalization and automation of the brewhouse operation at the Carlsberg Breweries in Copenhagen
Authors: Munksgaard Jannik 
Published: November 2023

Abstract
The aims of the modernization-project were several. It was intended to improve wort quality, improve the brewhouse yield, improve the efficiency and reduce the consumption of utilities. The project was undertaken under timerestrain, as full production had to be maintained throughout the rebuilding phase, which should be finalized before taking over production in connection with the general rationalization of Carlsberg’s Danish operation. The main items of the project were the installation of new mash-filters and a control system covering the whole brewhouse operation. However, the brewhouse was almost entirely renewed. The project succeeded on all expectations, though not without problems during the implementation.
Microbiological contaminations in immobilized yeast bioreactors for primary fermentation
Authors: Haikara A , Virkajarvi I , Kronlof J , Pajunen E 
Published: November 2023

Abstract
Microbiological purity in beer fermentation is a prerequisite, particularly in continuous fermentation. In contrast to secondary fermentation a wide range of micro-organisms was able to attach and grow in immobilized yeast bioreactors for primary fermentation. Wort bacteria with high specific growth rates were the most hazardous ones, producing off-flavours such as dimethyl sulphide. A Saccharomyces wild yeast caused both superattenuation and formation of phenolic off-flavours. Lactic acid bacteria were neither able to multiply nor to produce off-flavours. Aseptic treatment of wort and yeast, efficient monitoring also by means of chemical analysis and appropriate cleaning procedures are suggested for hindering microbiological contaminations.
Purification and characterization of yeast 3-methyl butanal reductases involved in the removal of wort carbonyls during fermentation
Authors: Ncdcrvelde L Van , Verlinden V , Philipp D , Debourg A 
Published: November 2023

Abstract
Aldehydes are known to impact negatively on beer flavour and flavour stability and yeast reductase activity is essential in the removal of the worty off-flavours during fermentation. In order to characterize the 3-methyl butanal reductases recently identified in yeast, a 3 steps purification procedure has been developed and the pure enzyme is identified by a specific detection after electrophoresis on acrylamide gel. The obtention of this purified enzyme has allowed a detailed study of the enzyme affinity and the determination of the N-terminal amino acid sequence is underway and will lead to the cloning of the gene encoding this enzyme.