bluuebunny/crossref_metadata_2025 huggingface dataset:
Reverse-phase chromatography of proteins
Authors: Corran P H
Published: November 2023

A glance through the literature shows that reverse-phase high-performance liquid chromatography (RP-HPLC) has now been assimilated into the canon of protein purification techniques: from the status of a novel, noteworthy technique when first applied it has now matured to that of being simply listed in the methods section along with high-performance size-exclusion, ion-exchange, or hydrophobic-interaction chromatography, no more noteworthy than any of the other techniques routinely employed or considered in any protein separation. The main way in which it differs from its companion techniques is in the necessary exposure of the sample proteins to high concentrations of organic solvents, and often low pH, both factors which can lead to the rapid denaturation of proteins, and which result in less denaturing methods being preferred, particularly where oligomeric proteins or enzymes are involved. Nevertheless, there are many circumstances where the risk of denaturation is not important; these include: most purely analytical work; separations for structural studies, or where structure and activity are retained or recovered despite the high concentrations of organic modifier; and where a rapid, simple, and powerful technique of unusual specificity like RP-HPLC is of great service.
Growth factors
Authors: Heath John K
Published: November 2023

Growth factors are extracellular polypeptide mediators of intercellular signalling. They exert their action by binding to specific receptors expressed by target cells, thereby initiating signal transduction processes inside the cell. Depending upon the nature of the target cell and the identity of the specific signalling mechanisms involved, changes in cell behaviour ensue. These can include the induction of cell differentiation, migration, death or survival, as well as activation or inhibition of cell multiplication (reviewed in ref. 1). The significance of growth factors in the context of on cogenesis is therefore that they have the property of evoking precisely those cellular responses that are subverted in the course of oncogenic transformation and tumorigenesis. Abnormal expression of growth factors, and consequential activation of cell signalling is therefore a potentially significant mechanism of oncogenic transformation.
RNA editing: posttranscriptional restructuring of genetic information
Authors: Hajduk Stephen L , Sabatini robert S
Published: November 2023

The formation of translatable mRNAs in the mitochondrion of kinetoplastid protozoa often requires the precise addition or deletion of uridines by RNA editing. The sites of uridine insertion and deletion are specified by small guide RNAs (gRNAs) which appear to function in directing the editing reactions. The extent of RNA editing can vary from the modest four uridines inserted into the cytochrome oxidase II mRNA, to the extensive editing seen in the cytochrome oxidase III mRNA of Trypanosoma brucei, in which hundreds of uridines are added and dozens are deleted. In most cases, the precursor mRNAs and the gRNAs needed for their editing are encoded on separate mitochondrial genomes. The gRNAs are encoded by numerous minicircles while the mRNAs are encoded by the less abundant maxicircles. Thus, genetic information from separate genomes is stitched together by the process of RNA editing. In this chapter, the possible molecular basis of RNA editing will be discussed. The biochemical mechanism and the ribonucleoprotein particles (RNPs) involved in these reactions are only now being resolved. The more complex issues of biological function and evolutionary origin of editing will also be considered. We have not attempted to review the editing literature exhaustively here (that has recently been done; see Refs. 1-7) but rather present the alternative views on mechanism, origins and function of what is a truly remarkable biological phenomenon.
History and overview
Authors: Baumberg Simon
Published: November 2023

This chapter sets the scene for the rest of the book by describing the early development of the subject and how the main themes were established. The choice of references reflects three not easily reconcilable aims: to enable the reader to follow how the topic developed and to find a way into recent literature (in conjunction with later chapters), and to give credit to at least some of the scientists principally involved. A brief mention, with selected references, is also given of systems which do not introduce novel mechanisms but which apply to prokaryotic properties of fundamental or applied importance.
Folding proteins
Authors: Rudolph Rainer , Böhm Gerald , Lilie Hauke , Jaenicke Rainer
Published: November 2023

The pioneering studies by Anson, Mirsky, Pauling, Anfinsen, Baldwin, and others paved the way for protein folding to become a major topic in biochemistry (1-3). In subsequent work, theoretical and mechanistic aspects of protein folding and, in recent years, applied and biological aspects were analysed in great detail (4-6).
Radiation hybrid mapping
Authors: Stewart Elizabeth a. , Cox david r.
Published: November 2023

Constructing a map of an area of interest has been one of the first endeavours of explorers throughout the ages. As the scientific community delves further into one of its newest areas of exploration, the human genome, fast and accurate methods of mapping are becoming increasingly important.
PCR3
Authors: Herrington C Simon , O’Leary John J
Published: November 2023

PCR in situ hybridization allows the detection of specific nucleic acid sequences and their distribution in the cell. It combines two powerful techniquesin situ hybridization (ISH), which allows cellular localization of DNA and RNA sequences in cells and tissues, and the polymerase chain reaction (PCR), which allows reproducible amplification of rare nucleic acid sequences. The combined technique and its variants greatly enhance the sensitivity of in situ hybridization and add morphological localization to the sensitivity of PCR. Such techniques have enormous potential for research and diagnosis but problems with reproducibility and reliability are often encountered. This book overcomes these problems by describing the key procedures in step-by-step detail and by providing the essential advice needed for success. Topics include: DNA in situ PCR and DNA PCR in situ hybridization (PCR ISH) for the detection of DNA targets in cells; reverse trancriptase in situ PCR (RT-PCR) and RT-PCR ISH for the detection of RNA targets; and PRINS (primed in situ synthesis) for chromosomal analysis in interphase nuclei and metaphase chromosome spreads. There are further chapters on fixation of tissues for PCR, selective ultraviolet radiation fractionation (SURF), application of in situ PCR to human tissues, applications and modifications of PCR-ISH, and automation of in situ amplification. PCR In Situ Hybridization is a unique and timely collection of well-tested protocols for the amplification of DNA and RNA in cells and tissues, drawing on the accumulated knowledge and experience of leading exponents of these techniques. For each topic covered, the authors provide detailed guidance on the key steps in the protocols, numerous hints and tips for success, and advice on trouble-shooting. PCR In Situ Hybridization will be invaluable to molecular biologists, pathologists, geneticists, and all those seeking to perform in situ analyses of nucleic acid molecules.
Protein detection methods
Authors: Merril Carl R , Washart Karen M
Published: November 2023

Advances in detection methods for proteins have paralleled the development of new separation techniques over the last three decades. Selection of the proper protein visualization procedure is as important as the separation technique employed in the analysis of proteins from complex mixtures. The ease of use, speed of detection, level of sensitivity, and integrity of the proteins following detection (i.e. to allow further processing of the proteins if necessary) are important criteria for determining the appropriate detection method. Suitable methods also need to maintain a linear and stoichiometric relationship between protein detection signal and protein concentration.
Video microscopy
Authors: Weiss Dieter G , Maile Willi , Wick Robert A , Steffen Walter
Published: November 2023

A new quality of microscopy, called video microscopy, emerges, if one observes the specimen, instead of with the human eye, with a video camera connected to video processing equipment working at real time. Video microscopy is, there fore, much more than just adding a camera and monitor to the microscope to share the images with a larger audience. More recently, electronic devices other than video cameras, such as high sensitivity charge coupled device (CCD) cameras and scanning light detector systems for confocal microscopy have been added to microscopes. The three fields (i) video-enhanced contrast microscopy for highest resolution work, (ii) video-intensified microscopy for low light applications, and (iii) electronic scanning microscopy for confocal microscopy and 3D imaging differ in the type of device generating the electronic image, but all three use basically the same types of analogue and digital image processors. While all these techniques are generally defined as electronic light microscopy, this chapter, video microscopy, deals with the first two techniques that involve CCD and video cameras as imaging devices. Video microscopy has produced a revolution in light microscopy of biological samples equivalent to that of the development of the immunofluorescence technique. It has once more made the traditional light microscope a powerful tool for those working on dynamic aspects of small biological systems, for example biochemists, molecular and cell biologists. It has given further resolving power to the light microscope enabling the observation of particles which bridge the size range between those normally studied by electron microscopy and those which are already well known to light microscopists as a whole, with the added advantage in that specimens can be examined alive. As well as allowing small particles to be resolved, the technique has the capacity to clean up the image, so allowing greater visibility. Also, changes of such parameters as amounts, concentrations, transport, or metabolism of specific molecules in both time and space can be quantitatively determined.
Reflection-contrast microscopy
Authors: Ploem J S , Prins F A , Velde I Cornelese-Ten
Published: November 2023

Fauré-Fremiet (1) used reflected light in cell biology studies half a century ago. Curtis (2) and Izzard and Lochner (3) performed the first fundamental studies of interference images of living cells on glass surfaces using reflected light microscopy. Reflection images were obtained with scanning reflecting microscopy (4, 5). Reflected-light microscopy for the study of living cells (2, 3, 6, 7) has been described as interference reflection, reflection-interference contrast, surface-contrast, and surface-:reflection interference microscopy (8). Ploem (9, 10) investigated further optical methods to improve the image contrast in reflected-light microscopy. In collaboration with Leica (11-13), an improved microscope system was developed for reflected-light microscopy. It used an aperture diaphragm with a central stop (creating an annular aperture) in the epi-illumination light path at an aperture plane conjugate with the back focal (aperture) plane of the objective (14). This was combined with epipolarization microscopy using immersion objectives equipped with a quarter lambda plate in their front lens (15). New high aperture objectives were developed for high resolution biological microscopy. This optically improved version of reflected-light microscopy was named reflection-contrast micro scopy (RCM).
Introduction of materials into living cells
Authors: Schöpke Christian , Fauquet Claude M
Published: November 2023

Since its inception a decade ago, particle bombardment, also known as the ‘Biolistic process’ (1), has developed into a method of considerable importance in many areas of biology (see Table 1). It has been especially successful in the genetic transformation of plants, where the presence of a cell wall poses a significant obstacle for the introduction of large molecules, including DNA. The purpose of the first part of this chapter is to illustrate the use of particle bombardment for the introduction of DNA into plant cells and the utilization of reporter genes to follow the fate of genetically transformed cells, using cassava (Manihot esculenta) as an example. The reader who is interested in more detailed information on particle gun types and on different applications of particle bombardment is referred to several recently published reviews (2-4).
Laboratory Protocols for Conditional Gene Targeting
Authors: Torres Raul M , Kühn Ralf
Published: November 2023

Conditional gene targeting is an emerging technique for introducing mutations into the genes of laboratory mice. This collection of protocols comes from one of the world's premier laboratories for gene targeting The Institute of Genetics in Cologne and is intended for newcomers to the field as well as more experienced researchers. Topics covered include: the Cre/loxP recombination system and gene targeting in ES cells; gene targeting strategies; Cre/loxP recombination system; Gene targeting vectors consideration and use of loxP; nonselectable modifications and removal of selection marker genes; large deletions; gene replacement; chromosomal translocations; targeted integrations; conditional gene modification; LoxP-containing transgenes; conditional gene targeting; working with ES cells; ES cell transfection overview; embryonic feeder cells; ES cells handling and use; ES cell culture and transfection; ES clone picking; identification of homologous recombinants; freezing ES clones; thawing ES clones; genomic DNA preparation; cre-mediated neomycin deletion in ES cells; generation of mice preparation; mouse breeding for ES cell injection or aggregation; injection needles and holders; blastocyst preparation/injection; embryo aggregation; blastocyst transfer;
The Xenopus oocyte expression system for the cDNA cloning and characterization of plasma membrane transport proteins
Authors: Yao* Sylvia Y M , Cassꝉ Carol E , Young* James D
Published: November 2023

Oocytes of Xenopus laevis, the South African clawed frog, represent a simple, but powerful system for the transient expression of heterologous proteins. These large and resilient cells have a high translational capacity and are readily microinjected with exogenous RNA or DNA. As a system for studying plasma membrane transport proteins, Xenopus oocytes are readily amenable to both radioisotope flux measurements and electrophysiological recordings. They also subject newly synthesized proteins to various forms of processing, including glycosylation and correct plasma membrane insertion. Unlike other whole-cell heterologous expression systems, Xenopus oocytes have a generally low background of endogenous transport activity, making them uniquely versatile with respect to the range of permeants and types of transporters that can be studied. The ability of Xenopus oocytes to translate foreign mRNA was first described in 1971 (1). Since then, the system has been applied with considerable success to the.study of receptors and channels and, following the landmark expressioncloning of the intestinal glucose transporter SGLTl (sodium-dependent glucose transporter 1) in 1987 (2), to transport proteins. In this chapter, we provide a laboratory guide to the Xenopus oocyte expression system for the cDNA cloning and functional and molecular characterization of plasma membrane transport proteins, illustrating some of these applications with our own studies of nucleoside transport processes in mammalian and other species. Other sources should be consulted for routine molecular biology techniques (3).
Human monoclonal antibodies to blood group antigens
Authors: Kumpel Belinda M
Published: November 2023

Many human alloantigens on red blood cells are not recognized by the mouse immune system, and therefore murine monoclonal antibodies to many desired blood group specificities cannot be produced using conventional murine hybrid oma technology. Among these is the Rh (rhesus) D antigen, which is the most immunogenic of the blood group alloantigens and clinically is the most import ant after the ABO blood groups. Most human monoclonal blood group antibodies have been made to the Rh D polypeptide. Monoclonal anti-D has been produced to replace or supplement plasma-derived anti-D for blood grouping (1) and for therapeutic Rh D prophylaxis for prevention of haemolytic disease of the new born (2). Also, there are more individuals with high titre anti-D than with anti bodies to other blood groups, who can donate source material. For this reason, the methods described in this chapter are those that have been used for the production of monoclonal anti-D.
Visualizing fluorescence in Drosophila–optimal detection in thick specimens
Authors: Davis Ilan
Published: November 2023

Drosophila has been a key model organism for studying the genetic control of development. Its success is largely due to its powerful genetics combined with the ability to visualize specific gene products in a beautiful multicellular cytology. This often involves detecting proteins and mRNA deep in whole-mount specimens such as embryos, egg chambers or imaginal discs. Such experiments can be technically challenging, since most objective lenses are designed to image sources of light located just below the coverslip. Imaging fluorescence at some distance from the coverslip creates a number of distortions which reduce the resolution and lower the signal-to-noise ratio.
Ras
Authors: Bos Johannes L
Published: November 2023

Ras is a molecular switch that controls the transduction of signals from cell-surface receptors to intracellular targets (Fig. la). Like all GTPases, the switch mechanism consists of activation by exchange of bound GDP for GTP, and inactivation by hydrolysis of GTP into GDP. Ras achieved notoriety in the 1980s with the discovery that 15% of all human tumours contain a point mutation in one of the three Ras genes. The resulting mutant protein is unable to hydrolyse bound GTP and therefore remains constitutively in the active conformation. As a consequence, cells experience a continuous growth-promoting signal in the absence of extracellular cues. Ras has been studied extensively and much is known about mechanisms of activation and of downstream signalling. In most cells, Ras is activated in response to a variety of extracellular signals, which stimulate tyrosine kinases either directly or indirectly. In its GTP-bound state, Ras associates with one of several effector molecules, most notably members of the Raf family, the RalGDS family, or PI3K. The biological consequences of these interactions depend greatly on the cell type and on the context of other signalling events. In addition, poorly understood parameters, such as the timing and intensity of the signal, may determine the ultimate response. Interestingly, Ras is highly conserved both in structure and function during evolution. In the yeast Saccharomyces cerevisiae, Ras is involved in nutrient response, in Schizosaccharomyces pombe Ras regulates the pheromone response in mating, and in the nematode Caenorhabditis elegans and the fruitfly Drosophila Ras is a key component of numerous developmental processes. This chapter will provide a background to Ras and present some current ideas about the molecular mechanisms of Ras signalling.
Synaptic vesicle endocytosis and recycling
Authors: Zhang Bing , Ramaswami Mani
Published: November 2023

‘The yang having reached its climax retreats in favor of the yin; the yin having reached its climax retreats in favor of the yang.’ -Wang Chu’ung (AD 80) on Tai Chi Tu Exocytosis and endocytosis, the yin and yang elements of the vesicle cycle, must be well balanced to ensure continuous synaptic transmission. At nerve terminals, the number of synaptic vesicles is insufficient to account for the number of quanta released during a short burst of intense nerve activity. Axonal transport of newly synthesized vesicle proteins occurs too slowly to refill the vesicle pool at the required rates. Under these circumstances, the only way to rapidly replenish synaptic vesicles is to recycle them locally through endocytosis. Thus, a major challenge is to understand how synaptic vesicles are faithfully and efficiently recovered.
Cyclin-dependent protein kinases
Authors: Gould Kathleen L
Published: November 2023

Cyclin-dependent protein kinases (Cdks) regulate progression through the eukaryotic cell cycle. They are also involved in the regulation of other processes such as gene expression and phosphate metabolism. Cdks are the catalytic subunits of hetero dimeric complexes and they approximate the size of a minimal protein-kinase catalytic domain (∼, 34 kDa). Alone, they lack protein kinase activity. As their name implies, By far the best understood of the Cdks are those that have important roles in cell cycle progression-Cdc2, Cdk2, and Cdk4. Cdks are activated transiently at particular stages of the cell cycle, and their activation or inactivation triggers the next events in the cycle. To ensure the coordination of these events, the activation of each Cdk is strictly regulated in a variety of ways including an elaborate cascade of protein phosphorylation events, interaction with subunits in addition to cyclins, and by regulated proteolysis of the cyclin and other subunits. Due to the wealth of knowledge concern ing Cdks and the limitations of space, it is impossible to include all pertinent information about Cdks in this chapter or to cite all the original references. Numerous recent reviews emphasize various aspects of Cdk function and regulation, and readers will be referred to these where appropriate.
Bacteriology of urine
Authors: Davies Eleri M , Lewis Deirdre A
Published: November 2023

The urinary tract above the level of the distal urethra is normally sterile but infection at any site within the tract is possible, from the kidney to the urethra, the commonest site of infection being the urinary bladder. In general microorganisms gain access to the urinary tract from neighbouring sites, most commonly the perianal region, resulting in ascending infection, although spread by the haematogenous route is usually responsible for infections by Salmonella spp., and Mycobacterium tuberculosis.
Antimicrobial assays
Authors: Reeves David S , Whitea Les O
Published: November 2023

An antibiotic was originally defined as a substance produced by a micro-organism which inhibited the growth of other micro-organisms. However many drugs loosely referred to as antibiotics are semi-synthetic derivatives of natural antibiotics (e.g. various penicillins and cephalosporins) or compounds produced solely by chemical means (e.g. sulfonamides, 4-quinolones). In addition there are true antibiotics which have useful anticancer rather than antimicrobial properties. For this reason it has become customary to refer to antimicrobial antibiotics and other antimicrobial drugs as antimicrobial agents, a term which is often reduced to antimicrobial. This terminology will be used in this chapter.
Early stages of protein folding
Authors: Roder Heinrich , ElÖve GÜlnur A , shastry m. C Ramachandra
Published: November 2023

Protein-folding reactions often appear deceptively simple. At equilibrium one can typically distinguish only two states, the precisely folded native state and a broad ensemble of largely featureless, unfolded conformations. Many small, globular proteins exhibit simple, first-order folding and unfolding kinetics that can be modelled as a reaction involving two states separated by a single free-energy barrier (reviewed in ref. 1). Any additional slow kinetic phases can often be attributed to heterogeneity of the unfolded state due to cis-trans isomerization of praline peptide bonds (2, 3) or to exchange of metal ligands (4-6).
Chromosome analysis and sorting by flow cytometry
Authors: Davies Derek C , Youngt Bryan D
Published: November 2023

Many types of cancer and genetic disease are characterized by chromosomal abnormalities. These can be detected by conventional cytogenetics, which involves photographing banded chromosomes on metaphase spreads. Although this is a widely used technique in haematology, oncology, and pre-natal diagnosis, it is a time-consuming process that relies heavily on the skill and experience of the cytogeneticist. In addition, in cancerous cells, the sometimes complex karyotypes encountered mean that such analysis is often extremely difficult. An alternative approach is to prepare a monodispersed suspension of chromosomes, stain with one or two fluorescent DNA dyes, and pass them through a flow cytometer. The intensity of the fluorescence signal from each chromosome is recorded, the values being dependent on their DNA content. In single-colour analysis, a non-base-specific dye such as ethidium bromide is used. Accumulating data from 50000 chromosomes and presenting this as a histo gram of fluorescence intensity produces a distinctive species-specific pattern of peaks. However, not all chromosomes appear as a single peak. An improvement is to use bivariate analysis that exploits the base-pair binding preferences of DNA specific dyes: usually Hoechst 33258 (which has an adenine/thymidine (AT) binding preference) in combination with chromomycin A3 (which has a guanine/cytosine (GC) binding preference). The intensity of staining with these fluorochromes is dependent not only on the DNA content but also on the base composition of each chromosome. The data accumulated can be presented as an isometric plot or, better, as a dot plot or contour map (see Figure 1). The bivariate plot resolves all the human chromosomes as separate peaks, with the exception of chromosomes 9, 10, 11, and 12.
Genetic analysis of basidiomycete fungi
Authors: Walser Piers J , Hollenstein Marcel , Klaus, Michaela J , Kües Ursula
Published: November 2023

The basidiomycetous fungi are divided into two main groups, the heterobasidiomycetes and the homobasidiomycetes. For the most part, genetic studies of heterobasidiomycetes have been restricted to Ustilago maydis, Microbotryum violaceae, and other smut fungi, while Coprinus cinereus and Schizophyllum commune have served as model organisms for the homobasidiomycetes. Other homobasidiomycetes are studied for industrial purposes, for example the white rot fungus Phanerochaete chrysosporium which produces exoenzymes such as laccases, and various edible mushrooms including Agaricus spp. These species can easily be cultured on relatively simple media, and they complete their entire life cycle within a few weeks under laboratory conditions. Since we are working with C. cinereus, here we present genetic techniques that are applied to this fungus. The approaches, however, are broadly applicable to the full range of basidiomycetes.
Methods in solid tumour cytogenetics
Authors: Mandahl N
Published: November 2023

It has been demonstrated beyond doubt that the acquired chromosome abnormalities, \vhich are a characteristic feature of human neoplastic cells, are non randomly distributed throughout the genome and that certain aberrations are associated with specific histological tumour types (1,2). These chromosomal changes, including both structural and numerical aberrations, affect all chromosomes, autosomes as well as gonosomes, and may be found as sole changes, but, particularly in solid tumours, more often in a complex array of karyotypic changes.
Plant histology
Authors: Spence Jackie
Published: November 2023

The microscope is one of the most important instruments used in biological research and it has contributed much to our knowledge of morphology and cell dynamics. Over the past ten years there have been many advances in light microscopical techniques including the development of the confocal microscope and improved image analysis software. However the need for simpler analytical techniques has certainly not decreased. Today molecular biologists and biochemists often require the light or fluorescence microscope to analyse gene expression patterns, mutant phenotypes, and structure-function relationships (1), and, although most laboratories have access to a simple light and fluorescence micro scope, not all laboratories have the expertise in microscopical techniques. The aim of this chapter is to introduce the non-microscopist to some of the more simple, but no less informative, histological techniques for the study of plant tissues.
Hsp70 chaperone networks: the role of regulatory co-chaperones in coordinating stress responses with cell growth and death
Authors: Song Jaewhan , Morimoto Richard I
Published: November 2023

Among the fascinating problems in biology is the process by which proteins acquire and maintain their folded state. Interest in this problem has intensified with the recognition that folding intermediates adopt alternative conformations rich in β-sheets that can self-associate to form protein aggregates, protofilaments, and fibrils as associated with a class of diseases known as protein conformational or aggregation diseases, including Alzheimer’s Disease, Parkinson’s Disease, ALS, and Huntington’s Disease (1, and see Chapter 11). As environmental and physiological stress and genetic mutations are known to negatively influence the stability of proteins and result in protein misfolding, it would not be surprising that the cellular stress response has an essential role in protein folding quality control.
High resolution FISH mapping using chromatin and DNA fibre
Authors: Heng Henry H Q
Published: November 2023

This chapter focuses on the method of high resolution fibre FISH. Fibre FISH collectively refers to all approaches that perform fluorescence in situ hybridization on released chromatin or DNA fibre. Since the degree of condensation is much less than metaphase chromosomes and even less than interphase chromatin, the resolution of fibre FISH is among the highest when compared to prophase chromosome FISH, interphase FISH, meiotic prophase FISH, or pronuclear FISH (1-5).
The American Lie
Authors: Ginsberg Benjamin
Published: November 2023

Going all the way back to the time of George Washington, much of what we see and hear in the political world consists of lies and deceptions. Despite assurances to the contrary, politics is not about truth, justice, and principle. It is about money, power, and status. As astute political commentator Ben Ginsberg convincingly demonstrates, politicians habitually lie, pretending to fight for principles, in order to conceal their true selfish motives. Citizens who need the frequent injunctions to participate in politics and abjure political cynicism are likely to be duped into contributing their tax dollars and even their lives for dubious purposes. Most individuals gain little from political participation. Participants are the foot soldiers of political warfare, but even if their side is victorious, they receive few of the spoils of war. Thus, in this new political season, Ginsberg encourages citizens to think outside the (ballot) box, finding new ways to act on behalf of their interests and the public good. But if they do vote, their motto should be when in doubt vote them out. The elections of 2008 are a good time to begin.
‘Drowned in a Sea of Inhumanity’: Natural Childbirth, Postnatal Depression and the National Childbirth Trust, 1956–80s
Authors: Marland Hilary
Published: November 2023

During the 1970s, the National Childbirth Trust (NCT) began to provide information and support to women experiencing postnatal mental illness, building on its promotion of natural childbirth and emphasis on the emotional wellbeing of women around birth, which had occupied the organisation since its establishment in 1956. This article argues that, alongside emotional, social and medical factors, the NCT attributed postnatal depression to the shift to hospital deliveries, involving high levels of intervention and frustrating women’s choice and agency. While sharing ambitions to improve care in childbirth and giving women a voice in describing their experiences, it is suggested that the NCT’s relationship with the feminist health movement remained ambiguous. The article also explores the NCT’s collaboration with a variety of experts and advisors, some of whom emphasised the risk of postnatal depression to the bonding process and infant’s development, potentially exacerbating the mental distress of new mothers.
Preliminary Validation of the Pain Relief Motivation Scales (PRMS)
Authors: Letzen Janelle E. , Hunt Carly A. , Webb Caroline , Vetter Maria , Finan Patrick H. , Karoly Paul , Mun Chung Jung
Published: November 2023

Objectives:

Pain typically prompts individuals to seek relief. This study aimed to develop and psychometrically validate the Pain Relief Motivation Scales (PRMS), applying revised Reinforcement Sensitivity Theory to measure the neuropsychological systems underlying motivation for pain relief. We hypothesized a 6-factor structure based on previous work, including one behavioral inhibition system (BIS) factor, one Fight-Flight-Freeze System (FFFS) factor, and four behavioral activation system (BAS) factors.

Methods:

Items were generated by adapting the Reinforcement Sensitivity Theory of Personality Questionnaire for relevance to pain relief. Adults with chronic pain were recruited internationally to participate in online survey batteries at baseline and one week later during 2021. We randomly split the sample to conduct exploratory factor analysis (n=253) and confirmatory factor analysis (n=253). Psychometric properties were estimated using the full sample (N=506).

Results:

Parallel analysis revealed that a 5-factor structure best fit the data (21 items): [1] hopelessness about pain relief (BIS), [2] hesitancy for engaging in pain treatments (BIS), [3] persistence in engaging in pain treatments (BAS), [4] relief reactivity (BAS), and [5] risky relief-seeking (BAS). Acceptable internal consistency (Cronbach’s alpha=.68-.80) and test-retest reliability (ICCs=.71-.88) were observed. Construct validity varied from weak to moderate (r’s=.02-.45).

Discussion:

As the first attempt to create an instrument measuring neuropsychological systems underlying motivation for pain relief, the findings show that additional work is needed to refine theory and psychometric rigor in this area. Cautiously, the results suggest that a BIS-BAS model, with minimal FFFS contributions, might be useful for understanding motivation for relief.
Longitudinal Pain Outcomes Following an Intensive Pediatric Pain Rehabilitation Program
Authors: Weiss Karen , Li Rui , Wang Xing , Vandeleur Daron , Havkins Sabina , Palermo Tonya M.
Published: November 2023

Objectives:

This study tested performance measures of physical functioning and pain-related psychosocial measures as potential mechanisms of improvements in outcomes following intensive outpatient interdisciplinary pain rehabilitation for adolescents. We hypothesized that improvements in performance measures of physical functioning, fear of pain, pain catastrophizing, and self-efficacy during treatment would be related to improvements in pain, functional disability, and depressive symptoms.

Methods:

Seventy-seven adolescents, 10-18 years old, completed self-report measures prior to treatment, after the first week, at discharge, and at 3-months follow-up. Participants completed physical performance measures of physical and exercise capacity and core strength on day one, after the first week, and at discharge. Linear mixed models were used to examine the association between changes in potential mechanisms and changes in outcomes, adjusting for age, sex, pain duration and baseline functional disability.

Results:

Statistically significant improvement in functional disability, pain, and depressive symptoms was demonstrated across the four time points. Improvements were also demonstrated in physical performance measures, fear of pain, pain catastrophizing and self-efficacy. Improvements in pain catastrophizing predicted improvements in functional disability, pain and depressive symptoms. Improvements in self-efficacy predicted improvements in all outcome variables except depression. Improvements in physical performance measures predicted improvements in some aspects of functional disability and pain, but not depressive symptoms.

Discussion:

This study contributes to the growing literature on the effectiveness of pediatric intensive interdisciplinary pain rehabilitation programs and mechanisms that influence improvements in outcomes, an area that is currently underexplored. Results are important to guide future research and inform clinical practice.
Sodium Bicarbonate and Calcium Chloride for the Treatment of Hyperkalemia-Induced Cardiac Arrest: A Randomized, Blinded, Placebo-Controlled Animal Study*
Authors: Eggertsen Mark Andreas , Munch Johannsen Cecilie , Kovacevic Alexander , Fink Vallentin Mikael , Mørk Vammen Lauge , Andersen Lars W. , Granfeldt Asger
Published: November 2023

OBJECTIVES:

Current international guidelines recommend administrating calcium chloride and sodium bicarbonate to patients with hyperkalemia-induced cardiac arrest, despite limited evidence. The aim of this study was to evaluate the efficacy of calcium chloride and sodium bicarbonate on return of spontaneous circulation (ROSC) in a pig model of hyperkalemia-induced cardiac arrest.

DESIGN:

A randomized, blinded, placebo-controlled experimental pig study. Hyperkalemia was induced by continuous infusion of potassium chloride over 45 minutes followed by a bolus. After a no flow period of 7 minutes, pigs first received 2 minutes of basic cardiopulmonary resuscitation and subsequently advanced life support. The first intervention dose was administered after the fifth rhythm analysis, followed by a defibrillation attempt at the sixth rhythm analysis. A second dose of the intervention was administered after the seventh rhythm analysis if ROSC was not achieved. In case of successful resuscitation, pigs received intensive care for 1 hour before termination of the study.

SETTING:

University hospital laboratory.

SUBJECTS:

Fifty-four female Landrace/Yorkshire/Duroc pigs (38–42 kg).

INTERVENTIONS:

The study used a 2 × 2 factorial design, with calcium chloride (0.1 mmol/kg) and sodium bicarbonate (1 mmol/kg) as the interventions.

MEASUREMENTS AND MAIN RESULTS:

Fifty-two pigs were included in the study. Sodium bicarbonate significantly increased the number of animals achieving ROSC (24/26 [92%] vs. 13/26 [50%]; odds ratio [OR], 12.0; 95% CI, 2.3–61.5; p = 0.003) and reduced time to ROSC (hazard ratio [HR] 3.6; 95% CI, 1.8–7.5; p < 0.001). There was no effect of calcium chloride on the number of animals achieving ROSC (19/26 [73%] vs. 18/26 [69%]; OR, 1.2; 95% CI, 0.4–4.0; p = 0.76) or time to ROSC (HR, 1.5; 95% CI, 0.8–2.9; p = 0.23).

CONCLUSIONS:

Administration of sodium bicarbonate significantly increased the number of animals achieving ROSC and decreased time to ROSC. There was no effect of calcium chloride on the number of animals achieving ROSC or time to ROSC.
Post-Cardiac Arrest Care in Adult Patients After Extracorporeal Cardiopulmonary Resuscitation
Authors: Kang Jin Kook , Darby Zachary , Bleck Thomas P. , Whitman Glenn J. R. , Kim Bo Soo , Cho Sung-Min
Published: November 2023

OBJECTIVES:

Extracorporeal cardiopulmonary resuscitation (ECPR) serves as a lifesaving intervention for patients experiencing refractory cardiac arrest. With its expanding usage, there is a burgeoning focus on improving patient outcomes through optimal management in the acute phase after cannulation. This review explores systematic post-cardiac arrest management strategies, associated complications, and prognostication in ECPR patients.

DATA SOURCES:

A PubMed search from inception to 2023 using search terms such as post-cardiac arrest care, ICU management, prognostication, and outcomes in adult ECPR patients was conducted.

STUDY SELECTION:

Selection includes original research, review articles, and guidelines.

DATA EXTRACTION:

Information from relevant publications was reviewed, consolidated, and formulated into a narrative review.

DATA SYNTHESIS:

We found limited data and no established clinical guidelines for post-cardiac arrest care after ECPR. In contrast to non-ECPR patients where systematic post-cardiac arrest care is shown to improve the outcomes, there is no high-quality data on this topic after ECPR. This review outlines a systematic approach, albeit limited, for ECPR care, focusing on airway/breathing and circulation as well as critical aspects of ICU care, including analgesia/sedation, mechanical ventilation, early oxygen/Co
2, and temperature goals, nutrition, fluid, imaging, and neuromonitoring strategy. We summarize common on-extracorporeal membrane oxygenation complications and the complex nature of prognostication and withdrawal of life-sustaining therapy in ECPR. Given conflicting outcomes in ECPR randomized controlled trials focused on pre-cannulation care, a better understanding of hemodynamic, neurologic, and metabolic abnormalities and early management goals may be necessary to improve their outcomes.

CONCLUSIONS:

Effective post-cardiac arrest care during the acute phase of ECPR is paramount in optimizing patient outcomes. However, a dearth of evidence to guide specific management strategies remains, indicating the necessity for future research in this field.
Tear Tacrolimus Levels and Clinical Response After Adjunct Therapy With Cutaneous Application of Tacrolimus 0.1% Over Upper Eyelid Skin in Chronic Vernal Keratoconjunctivitis
Authors: Bardoloi Priyadarshana , Vanathi Murugesan , Velpandian Thirumurthy , Laxmi Moksha , Gupta Noopur , Lomi Neiwete , Tandon Radhika
Published: November 2023

Purpose:

The aim of this study was to evaluate the role of cutaneous application of 0.1% tacrolimus eye ointment over the skin of the upper eyelid in chronic vernal keratoconjunctivitis (VKC).

Methods:

A prospective, longitudinal, noncomparative, open-label clinical study of moderate-to-severe grade steroid-dependent VKC was performed. Study participants were initiated on adjunct therapy of cutaneous application of 0.1% tacrolimus ointment twice daily on the upper eyelid skin. Ocular surface evaluation parameters, meibomian gland imaging, intraocular pressure, visual acuity, and clinical disease severity scoring were performed to assess clinical response at baseline and month 3 of therapy. Tear levels of tacrolimus were measured at month 3 using high-performance liquid chromatography tandem mass spectrometry and correlated with the clinical score.

Results:

Palpebral form of VKC was observed in 85% of the cases, with positive family history in 5%, atopy in 7.5%, and keratoconus in 11.25%. Clinical assessment revealed improvement in 97.5% patients with discontinuation of concomitant topical steroids in 64% of patients. There were no changes in visual acuity, intraocular pressure, or ocular surface evaluation after therapy. Tacrolimus was detected in the tears of all our study patients after cutaneous application over the upper eyelid skin, proving its bioavailability with mean tear tacrolimus levels of 6.55 ± 21.43 ng/mL. Correlation analysis revealed a moderate negative correlation between the clinical score and tacrolimus concentration (Spearman correlation coefficient: −0.34, P = 0.002).

Conclusions:

Cutaneous tacrolimus 0.1% ointment over the upper eyelid skin is an efficacious alternative method of application in treatment of VKC, with no resultant ocular irritation.
Understanding racial disparities of glaucoma
Authors: Barquet-Pizá Viviana , Siegfried Carla J.
Published: November 2023

Purpose of review

Increased prevalence, earlier onset, and more rapid progression to vision loss from glaucoma has demonstrated racial disparity in numerous studies over decades. Precise etiologies of these important differences among patients of African and Hispanic ancestral background have not been elucidated. This review focuses on currently available epidemiologic/population, genetic, socioeconomic and physiologic studies of racial disparities in this blinding disease.

Recent findings

In depth reviews of several landmark studies of glaucoma prevalence in various racial groups have highlighted potential challenges of lack of recruitment of diverse populations in genetic studies and clinical trials, challenges of racial stratification of subjects, and the impact of socioeconomic variables.

Through a more comprehensive analysis of racial disparities of glaucoma, both clinicians and researchers may provide more effective population screening and management with a holistic approach for individualized patient care to provide improved outcomes. Future studies of interventions in sociodemographic factors and genetic/physiologic variables that influence the prevalence, access, and consequential vision loss from glaucoma will be crucial to minimize/eliminate racial disparities and improve outcomes for all.
Prediction of Coronary Artery Lesions in Patients With Recurrent Kawasaki Disease
Authors: Yang Penghui , Zhang Jing , Liu Yihao , Feng Siqi , Yi Qijian
Published: November 2023

Background:

A subset of patients with Kawasaki disease (KD) will suffer recurrence. However, there is still a lack of accurate prediction models for coronary artery lesions (CAL) in recurrent KD patients. It is necessary to establish a new nomogram model for predicting CAL in patients with recurrent KD.

Methods:

Data from patients with recurrent KD between 2015 and 2021 were retrospectively reviewed. After splitting the patients into training and validation cohorts, the least absolute shrinkage and selection operator was used to select the predictors of CAL and multivariate logistic regression was used to construct a nomogram based on the selected predictors. The application of area under the receiver operating characteristic curve (AUC), calibration curves, Hosmer–Lemeshow test, Brier score and decision curve analysis were used to assess the model performance.

Results:

A total of 159 recurrent KD patients were enrolled, 66 (41.5%) of whom had CAL. Hemoglobin levels, CAL at the first episode, and intravenous immunoglobulin resistance at recurrence were identified by the least absolute shrinkage and selection operator regression analysis as significant predictors. The model incorporating these predictors showed good discrimination (AUC, 0.777) and calibration capacities (Hosmer–Lemeshow P value, 0.418; Brier score, 0.190) in the training cohort. Application of the model to the validation cohort yielded an AUC of 0.741, a Hosmer–Lemeshow P value of 0.623 and a Brier score of 0.190. The decision curve analysis demonstrated that the nomogram model was clinically useful.

Conclusions:

The proposed nomogram model could help clinicians assess the risk of CAL in patients with recurrent KD.
Differences in Clinical Presentation of COVID-19 in Children Hospitalized During Domination of Early (BA.1, BA.2) and Late (BA.5, BA.2.75, BQ.1 and XBB.1.5) SARS-CoV-2 Omicron Subvariants
Authors: Pokorska-Śpiewak Maria , Pawłowska Małgorzata , Ciechanowski Przemysław , Peregrym Michał , Dobrzeniecka Anna , Sobolewska-Pilarczyk Małgorzata , Franczak Justyna , Majda-Stanisławska Ewa , Szczepańska Barbara , Zaleska Izabela , Flisiak Robert
Published: November 2023

This study analyzed data for 1098 children: 575 diagnosed with COVID-19 between January and June 2022 (early Omicron) and 523 hospitalized from July 2022 to April 2023 (late Omicron). New Omicron subvariants lead to similar recovery rates without deaths and acute respiratory distress syndrome in children as BA.1 and BA.2, however, they more often cause fever and croup. Children suffering from comorbidities, presenting with pulmonary lesions and older, may be prone to a more severe consequences of COVID-19 in terms of the currently dominating Omicron subvariants.