bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sciatic nerve palsy secondary to postoperative haematoma in primary total hip replacement
Authors: Butt A. J. , McCarthy T. , Kelly I. P. , Glynn T. , McCoy G. 
Published: October 2005

Sciatic nerve palsy is a recognised complication of primary total hip replacement. In our unit this complication was rare with an incidence of < 0.2% in the past ten years. We describe six cases of sciatic nerve palsy occurring in 355 consecutive primary total hip replacements (incidence 1.69%). Each of these palsies was caused by post-operative haematoma in the region of the sciatic nerve. Cases, which were recognised early and surgically-evacuated promptly, showed earlier and more complete recovery. Those patients for whom the diagnosis was delayed, and who were therefore managed expectantly, showed little or no recovery. Unexpected pain and significant swelling in the buttock, as well as signs of sciatic nerve irritation, suggest the presence of haematoma in the region of the sciatic nerve. It is, therefore, of prime importance to be vigilant for the features of a sciatic nerve palsy in the early post-operative period as, when recognised and treated early, the injury to the sciatic nerve may be reversed.
Expression of vascular endothelial growth factor (VEGF) and its receptors in thyroid carcinomas of follicular origin: a potential autocrine loop
Authors: Vieira Joaquim Miguel , Santos Susana C Rosa , Espadinha Carla , Correia Isabel , Vag Tibor , Casalou Cristina , Cavaco Branca Maria , Catarino Ana Luiísa , Dias Sérgio , Leite Valeriano 
Published: October 2005

Objective: The aim of this study was to clarify the role of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) pathways in thyroid tumourigenesis.
Methods: We examined VEGF, VEGFR-1 and VEGFR-2 expression on 34 papillary thyroid carcinomas (PTCs), 18 follicular thyroid carcinomas (FTCs), eight poorly differentiated thyroid carcinomas (PDTCs) and on a thyroid tumour-derived cell line (NPA′87) by immunohistochemistry, reverse transcriptase PCR, immunofluorescence and Western blotting.
Results: We have demonstrated that VEGF expression was significantly (P < 0.05) more prevalent in PTCs (79%) than in FTCs (50%) or PDTCs (37%). Similarly, 76% of PTCs, 83% of FTCs and 25% of PDTCs expressed VEGFR-1, whereas 68% of PTCs, 56% of FTCs and 37% of PDTCs expressed VEGFR-2. Coexpression of VEGF and its receptors was observed in 50% of PTCs, 39% of FTCs and 12% of PDTCs, raising the possibility that VEGF may signal in an autocrine loop in these neoplasias, as observed previously for other types of cancer. In agreement with the idea that autocrine VEGF signalling plays an important role in thyroid carcinogenesis, the blockade of either VEGF or its receptors with neutralizing antibodies significantly reduced cell viability and increased apoptosis levels of the VEGFR-positive thyroid tumour cell line NPA′87.
Conclusions: Our results highlight a previously undefined VEGF autocrine action in thyroid carcinomas which could play a crucial role in tumour cell survival and could represent a useful therapeutic target for thyroid tumours.
Thyroid hormone increases mannan-binding lectin levels
Authors: Riis Anne Lene Dalkjær , Hansen Troels Krarup , Thiel Steffen , Gravholt Claus Højbjerg , Gjedde Signe , Gormsen Lars Christian , Jørgensen Jens Otto Lunde , Weeke Jørgen , Møller Niels 
Published: October 2005

Background: Recent studies have indicated the existence of causal links between the endocrine and immune systems and cardiovascular disease. Mannan-binding lectin (MBL), a protein of the innate immune system, may constitute a connection between these fields.
Methods: To test whether thyroid hormone regulates MBL levels, we studied eight patients with Graves’ hyperthyroidism before and after methimazole therapy, eight healthy subjects before and after short-term experimental hyperthyroidism, and eight hypothyroid patients with chronic auto-immune thyroiditis before and after L-thyroxine substitution.
Results: In all hyperthyroid patients, MBL levels were increased – median (range), 1886 ng/ml (1478–7344) – before treatment and decreased to 954 ng/ml (312–3222) after treatment (P = 0.01, paired comparison: Wilcoxon’s signed ranks test). The healthy subjects had MBL levels of 1081 ng/ml (312–1578). Administration of thyroid hormones to these persons induced mild hyperthyroidism and increased MBL levels significantly to 1714 ng/ml (356–2488) (P = 0.01). Two of the eight hypothyroid patients had undetectably low levels of MBL both before and after L-thyroxine substitution. The other six hypothyroid patients had decreased levels of MBL of 145 ng/ml (20–457) compared with 979 ng/ml (214–1533) after L-thyroxine substitution (P = 0.03, paired comparison: Wilcoxon’s signed ranks test).
Conclusion: Our data show that thyroid hormone increases levels of MBL. MBL is part of the inflammatory complement system, and this modulation of complement activation may play a role in the pathogenesis of a number of key components of thyroid diseases.
The expression of wild-type pendrin (SLC26A4) in human embryonic kidney (HEK 293 Phoenix) cells leads to the activation of cationic currents
Authors: Dossena Silvia , Maccagni Antonella , Vezzoli Valeria , Bazzini Claudia , Garavaglia Maria Lisa , Meyer Giuliano , Fürst Johannes , Ritter Markus , Fugazzola Laura , Persani Luca , Zorowka Patrick , Storelli Carlo , Beck-Peccoz Paolo , Bottà Guido , Paulmichl Markus 
Published: October 2005

Objective: The SLC26A4 protein (pendrin) seems to be involved in the exchange of chloride with other anions, therefore being responsible for iodide organification in the thyroid gland and the conditioning of the endolymphatic fluid in the inner ear. Malfunction of SLC26A4 leads to Pendred syndrome, characterized by mild thyroid dysfunction often associated with goiter and/or prelingual deafness. The precise function of the SLC26A4 protein, however, is still elusive. An open question is still whether the SLC26A4-induced ion exchange mechanism is electrogenic or electroneutral. Recently, it has been shown that human pendrin expressed in monkey cells leads to chloride currents.
Methods: We overexpressed the human SLC26A4 isoform in HEK293 Phoenix cells and measured cationic and anionic currents by the patch-clamp technique in whole cell configuration.
Results: Here we show that human pendrin expressed in human cells does not lead to the activation of chloride currents, but, in contrast, leads to an increase of cationic currents.
Conclusion: Our experiments suggest that the SLC26A4-induced chloride transport is electroneutral when expressed in human cellular systems.
Do You Hear Voices? A Study of the Symbolic Reading Inventory
Authors: Baer Allison L. 
Published: October 2005

In an effort to make reading more visible and find out what goes on in the minds of adolescent readers, the author led a group of seventh‐grade students in creating Symbolic Reading Inventories. The students created “snapshot” scenes from the books they were reading and placed themselves, as readers, in or around the picture.Reading is a complex puzzle made up of myriad pieces, some that fall easily into place and others that struggle to find some kind of fit. To make matters even more interesting, reading is an invisible process, deeply personal between the reader and the text. In interviewing chosen students, the author found that, in general, they were able to suspend reality and actively place themselves in the created scenes. In particular, the students engaged their imaginations, entering the text as they heard, saw, and felt aspects from chosen scenes. Not only did they bring life to the text, but they also brought their own lives to the text as they were actively engaged in creating meaning. Based on this information, the author encourages educators to provide situations that move our students from the commonplace to the extraordinary.
A Homogeneous Assay to Assess GABA Transporter Activity
Authors: Kopec Karla K. , McKenna Beth Ann , Pauletti Daniel 
Published: October 2005

AbstractThis unit describes a convenient functional uptake assay for GABA transport into cell lines transiently transfected with GABA transporter‐1 (GAT‐1) and other GAT isoforms. This facile, homogeneous assay allows for the determination of Km, Vmax, and Ki values. The assay utilizes commercially available microtiter plates that contain scintillant embedded in the bottom of the wells. Whereas a signal is generated as the cell accumulates the labeled neurotransmitter, label in the medium is undetected. While GABA uptake is observed in several cell lines transfected with GAT‐1, Km values for GABA uptake may vary with the cell line. This indicates that the choice of cell line is an important consideration when conducting uptake assays.
Adenylyl and Guanylyl Cyclase Assays
Authors: Kelly John J. , Stevens Troy , Thompson W. Joseph , Seifert Roland 
Published: October 2005

AbstractThis unit presents two basic protocols to determine adenylyl cyclase and guanylyl cyclase activity in tissue and cell homogenates, permeabilized cells, or subcellular fractions. Each method is divided into two parts: the enzyme reaction that causes the formation of the labeled cyclic nucleotide, and the separation of cyclic nucleotide products from unreacted nucleotide triphosphates and metabolites using Dowex 50 resin and aluminum oxide chromatographies. In the case of guanylyl cyclase, alternative separation protocols are also provided. Additionally, protocols are provided that describe preparation of both the columns used in the assays and the tissue or cells to be assayed.
Assay of Receptor‐Stimulated Phosphoinositide Turnover
Authors: Kendall David A. , Alexander Stephen P.H. 
Published: October 2005

AbstractThe stimulation of phosphoinositide turnover is one of the key means by which receptors evoke responses in target cells and tissues. This is true for both G protein‐coupled receptors and receptors that couple via tyrosine kinase activity. The protocols in this unit allow for pharmacological analysis of receptors coupled to phosphoinositide turnover. In general, the [3H]myo‐inositol prelabeling methodology (described for both tissue slices and cultured cells) is the more widely applicable, since it requires fewer experimental steps and typically gives rise to a better signal‐to‐noise ratio. Individual inositol phosphates can also be determined as described by chromatographic separation on ion‐exchange columns. In some circumstances (for example, when rapid responses to receptor stimulation are to be investigated or when the absolute levels of the active inositol phosphate are to be examined), it is preferable to use the mass assay described here for inositol (1,4,5)‐trisphosphate from either tissue slices and cultured cells. This unit also provides support protocols for the preparation of [3H]myo‐inositol, chromatography columns, tissue slices, and the IP3‐binding protein.
Overview of Pharmacokinetics
Authors: Aimone Lisa D. 
Published: October 2005

AbstractIn addition to having potency against the molecular target, a compound must be able to reach its intended site of action in vivo in sufficient quantity and for the appropriate duration to exert a biological effect. The fate of a compound after in vivo administration depends upon the absorption, distribution, metabolism, and excretion (ADME) of the compound, and the concentration of the compound in the blood, plasma, or other tissues represents the sum of all of these processes. Described in this unit are protocols for administering a compound by a variety of routes and for collecting the appropriate samples to determine the pharmacokinetic profile. The basic terms used in pharmacokinetic studies are defined, and representative examples are given to illustrate important variables to consider.
Large‐Scale Preparation of Conjugated Oligonucleoside Phosphorothioates by the High‐Efficiency Liquid‐Phase (HELP) Method
Authors: Bonora Gian Maria 
Published: October 2005

AbstractA new process for the preparation of large amounts of thioated oligonucleotides in a quasi‐classical solution condition is described. This method takes advantage of the use of polyethylene glycol as a soluble, inert support during synthesis. The easy purification of intermediates from a moderate excess of reagents allows very high coupling yields and, consequently, efficient production of the long oligonucleotide sequences required for pharmacological applications. The reaction of properly protected and activated oligonucleotides with high‐molecular‐weight polyethylene glycols allows a convenient procedure for the postsynthetic conjugation of those biopolymers in solution. The oligonucleotides are modified at the 5′ terminus using a liquid‐phase procedure with a linker carrying a terminal primary amino group to enhance its nucleophilic reactivity. The use of N, N′‐disuccinimidyl carbonate for the activation of the terminal OH groups of the PEG was preferred.
Synthesis of Covalent Oligonucleotide‐Streptavidin Conjugates and Their Application in DNA‐Directed Immobilization (DDI) of Proteins
Authors: Wacker Ron , Niemeyer Christof M. 
Published: October 2005

AbstractSemisynthetic DNA‐streptavidin conjugates are synthesized by covalent coupling of thiol‐modified DNA oligonucleotides and streptavidin (STV). The resulting conjugates have binding capacities for four equivalents of biotin and a complementary nucleic acid sequence. The conjugates are purified to homogeneity by ultrafiltration and chromatography, and are characterized by spectrophotometry and gel electrophoresis. Subsequently, the conjugates are applied as molecular connectors in the DNA‐directed immobilization (DDI) of biotinylated antibodies using DNA microarrays as immobilization matrices. The results are protein microarrays that can be used for the multiplexed detection of various antigens.
Synthesis of Acyclic Analogs of Adenosine
Authors: Holý Antonin 
Published: October 2005

AbstractThis unit describes preparation of two groups of chiral acyclic nucleosides that are N‐alkyl derivatives of adenine with hydroxyl group(s) on the alkyl chain. As an example of the neutral group, the synthesis is presented for (S)‐(2,3,dihydroxypropyl)adenine (DHPA), a compound with multiple biological effects in proliferating systems. Its synthesis consists of alkylation of adenine with a suitable chiral synthon based on 1,2‐O‐isopropylideneglycerol. Another group of structurally related biologically active compounds are the 9‐alkyladenines bearing both hydroxyl and carboxyl functions. Large‐scale cyanohydrin reaction of the adenine‐containing acetaldehyde intermediate yields 3‐(adenin‐9‐yl)‐2‐hydroxypropanoic acid (AHPA). Lastly, one of the alternative syntheses of its homolog, the nucleoside antibiotic eritadenine (with a d‐erythro‐2,3‐dihydroxybutanoic acid side‐chain) is described starting from d‐ribose. Procedures for preparation of all intermediates as well as the purification of the resulting crude material are described in detail. All these adenine derivatives inhibit SAM‐dependent methylation reactions via inhibition of SAH hydrolase.
Synthesis of Acyclic Nucleoside Phosphonates
Authors: Holý Antonin 
Published: October 2005

AbstractAcyclic nucleoside phosphonates (ANPs) are important biologically active nucleotide analogs. They contain an isopolar phosphonomethyl function linked to the hydroxyl group of an acyclic side‐chain via an undegradable ether group. Though their most important activity is antiviral, some exhibit cytostatic or antiprotozoic effects. The three most important groups of ANP are presented here as synthetic procedures for a large laboratory scale. Synthesis follows three principles: (1) introduction of a protected phosphonomethyl group to the hydroxyl on an appropriate alkyl side‐chain of an acyclic nucleoside, (2) alkylation of the heterocyclic base by a synthon with all characteristic features of the future phoshonate‐bearing side‐chain, or (3) transformation of a reactive group at the heterocyclic base. The last step in all these cases is removal of the phosphonate esters. Preparation methods are described in detail for PMEA, PMEG, PMEDAP and its N6‐cyclopropyl derivative, (R)‐PMPA, and (S)‐HPMPA, as well as all intermediates and synthons.
In Vivo Marking of Single Cells in Chick Embryos Using Photoactivation of GFP
Authors: Stark D. A. , Kulesa P. M. 
Published: October 2005

AbstractSelective marking of a single cell within a living embryo is often difficult due to the inaccuracy and invasiveness of standard techniques. This unit describes a minimally invasive optical protocol that uses 405‐nm laser light to photoactivate a variant of green fluorescent protein (PAGFP). This method takes advantage of the accessibility of the chick embryo to inject PAGFP into a region of interest and uses electroporation to deliver the construct into cells. This unit describes in detail how single and small groups of cells (n<10) that express PAGFP can be made visually distinguishable from the host population using the photoactivation process. Included is a means to maximize the fluorescence increase due to photoactivated GFP signal and to reduce photobleaching. Briefly outlined are previously developed chick culture and time‐lapse imaging techniques to allow for the subsequent monitoring of photoactivated cell migratory behaviors. The technique has the potential to be a less‐invasive, accurate tool for in vivo studies that involve following cell lineage and cell migration.
The Detection of MAPK Signaling
Authors: Shaul Yoav , Seger Rony 
Published: October 2005

AbstractMitogen‐activated protein kinase (MAPK) cascades are central pathways that participate in the intracellular transmission of extracellular signals. Each of the MAPK signaling cascades seems to consist of three to five tiers of protein kinases that sequentially activate each other by phosphorylation. Since the majority of MAPK cascade components are kinases, the methods used to detect their activation involve determining phosphorylation state and protein kinase activities. The Basic Protocol describes the use of immunoblotting with specific anti‐phospho antibody to detect activation of MAPK components. Alternative methods described are immunoprecipitation of desired protein kinases followed by phosphorylation of specific substrates and the use of an in‐gel kinase assay. These methods have proven useful in the study of the MAPK signaling cascades.
Stem Cells: An Overview
Authors: Denham Mark , Conley Brock , Olsson Fredrik , Cole Timothy J. , Mollard Richard 
Published: October 2005

AbstractStem cells are specialized cells that possess a capacity to undergo self‐renewal while at the same time having the ability to give rise to at least one or more differentiated or mature cell type. They therefore represent a fundamental cornerstone during the life of all vertebrates, playing central roles in the production of new and replacement cells for tissues during development and homeostasis, including repair following disease or injury. This unit is a review of stem cells, their roles in development, and their potentials as therapeutic agents.
Mouse Embryonic Stem Cell Derivation, and Mouse and Human Embryonic Stem Cell Culture and Differentiation as Embryoid Bodies
Authors: Conley Brock J. , Denham Mark , Gulluyan Lerna , Olsson Fredrik , Cole Timothy J. , Mollard Richard 
Published: October 2005

AbstractEmbryonic stem (ES) cells are pluripotent cells derived from developing mouse blastocysts in vitro that maintain long‐term self renewal and the capacity to give rise to all cell types in the adult body (including some extraembryonic cell types) when subjected to the appropriate conditions. It is envisaged that the development of methods enabling controlled differentiation of mouse ES cell counterparts from human blastocysts would enable the provision of an unlimited supply of tissue for cell and tissue transplantation therapies for the repair and replacement of diseased, injured, and senescent tissue. Furthermore, derivation of mouse ES cells has allowed for the generation of thousands of gene‐targeted mouse mutants. Culture of mouse ES cells as embryoid bodies (EBs) has provided a convenient system for studying early mouse developmental processes, including several aspects of extraembryonic lineage and axis formation associated with the pre‐ and peri‐gastrulating mouse embryo. Relatively little is known regarding the corresponding development of the early human embryo due to limitations associated with the acquisition of relevant tissue material for study. The transfer of methods such as EB formation to human systems should, by association, facilitate a more advanced understanding of similar processes associated with early human development. This unit describes protocols for isolating mouse embryonic stem cells and methods for propagating, freezing, and producing EBs from both mouse and human embryonic stem cells.
Using the KEGG Database Resource
Authors: Aoki Kiyoko F. , Kanehisa Minoru 
Published: October 2005

AbstractKEGG (Kyoto Encyclopedia of Genes and Genomes) is a bioinformatics resource for understanding the functions and utilities of cells and organisms from both high‐level and genomic perspectives. It is a self‐sufficient, integrated resource consisting of genomic, chemical, and network information, with cross‐references to numerous outside databases, containing a complete set of building blocks (genes and molecules) and wiring diagrams (interaction networks) to represent cellular functions. This unit describes protocols for using KEGG, KEGG PATHWAY, KEGG GENES, KEGG SSDB, KEGG EXPRESSION, and KEGG LIGAND. Protocols are also described for how to color maps, compare chemical compounds and glycan chains, analyze Ortholog Clusters, and visualize and analyze microarray data, among other procedures. All of these many protocols enable the user to take advantage of the full breadth of the functionality provided by KEGG.
Representing Structural Information with RasMol
Authors: Goodsell David S. 
Published: October 2005

AbstractThis unit describes the use of RasMol, a popular interactive molecular graphics program, for the display of biomolecular structures. The unit begins with a general guide to obtaining molecular coordinates from the Protein Data Bank and displaying them with RasMol, and then describes several molecular representations that are useful for different applications, along with methods for ensuring that one is viewing the appropriate biological unit and some ideas for customizing one's molecular graphics session.
Generating a Genome Assembly with PCAP
Authors: Huang Xiaoqiu , Yang Shiaw‐Pyng 
Published: October 2005

AbstractThis unit describes how to use the Parallel Contig Assembly Program (PCAP) to assemble the data produced by a whole‐genome shotgun sequencing project. We present a basic protocol for using PCAP on a multiprocessor computer in a 300‐Mb genome assembly project. A support protocol to prepare input files for PCAP is also described. Another basic protocol for using PCAP on a distributed cluster of computers in a 3‐Gb genome assembly project is presented, in addition to suggestions for understanding results from PCAP.
Laboratory Maintenance of Brucella abortus
Authors: Sun Yao‐Hui , den Hartigh Andreas B. , Tsolis Renée M. , Ficht Thomas A. 
Published: October 2005

AbstractThis unit provides protocols for growth of Brucella abortus on solid or in liquid media and for long‐term storage of laboratory stocks. Two issues affecting the culture and storage of isolates of this slow‐growing bacterium are emphasized: contamination of cultures and outgrowth of attenuated variants lacking a complete lipopolysaccharide. Laboratories planning to work with B. abortus should be aware that Biosafety Level 3 facilities are required. Furthermore, this organism is classified in the U.S. as a Select Agent, which restricts its use to laboratories registered with the U.S. government's Select Agent programs.