gebhart/patrepeval huggingface dataset:
Single‐fluorophore imaging with an unmodified epifluorescence microscope and conventional video camera (1999)

Single fluorophores in aqueous solution were imaged in real time with a conventional silicon-intensified target video camera connected to an unmodified commercial microscope (IX70, Olympus) with epifluorescence excitation with a high-pressure mercury lamp. Neither a powerful laser nor an extremely sensitive video camera was required. Three experimental systems were used to demonstrate quantitatively that individual, moving or stationary Cy3 fluorophores could be imaged with the microscope: Cy3-gelsolin attached to an actin filament sliding over heavy meromyosin, sliding actin filaments sparsely labelled with Cy3, and heavy meromyosin labelled with one or two Cy3 fluorophores. The results should encourage many laboratories to attempt 'single-molecule physiology' in which the functions and mechanisms of molecular machines are studied at the single-molecule level in an environment where the biological machines are fully active.
Modified vaccinia virus Ankara undergoes limited replication in human cells and lacks several immunomodulatory proteins: implications for use as a human vaccine. (1998)

Modified virus Ankara (MVA) is a vaccinia virus (VV) strain that was attenuated by serial passage through chick embryo fibroblasts (CEFs) and contains six large genomic deletions compared with parental virus. MVA replicates well in CEFs, but poorly in most mammalian cells. Recombinant MVA is a promising human vaccine candidate due to its restricted host range, immunogenicity and avirulence in animal models, and excellent safety record as a smallpox vaccine. Here we present a further characterization of MVA and demonstrate that: (i) MVA can replicate, albeit poorly, in transformed human cell lines, but not in primary human fibroblasts although there is limited cell-to-cell spread; (ii) MVA is a potent inducer of type I interferon (IFN) from primary human cells, which may restrict virus spread in vivo; and (iii) unlike other VV strains, MVA does not express soluble receptors for IFN-gamma, IFN-alpha/beta, tumour necrosis factor and CC chemokines, but does express a soluble interleukin-1beta receptor. This provides a plausible and testable explanation for the good immunogenicity of MVA despite its poor replication in mammals. The implications of these findings for the use of MVA as a safe and immunogenic human vaccine candidate are discussed.
Exogenous and endogenous corticosteroids modulate blood-brain barrier development in the ovine fetus (2000)

We previously reported decreases in blood-brain barrier permeability in the ovine fetus at 80% of gestation after antenatal corticosteroids and shown that permeability is not reduced in newborn lambs after postnatal corticosteroids. We now test the hypotheses that exogenous antenatal corticosteroids decrease blood-brain barrier permeability at 60% but not 90% of gestation in ovine fetuses and that endogenous increases in plasma cortisol concentrations are associated with ontogenic decreases in barrier permeability during gestation. Chronically instrumented ovine fetuses were studied 12 h after the last of four 6-mg dexamethasone or placebo injections were given 12 h apart over 48 h to ewes. Fetuses at 80% of gestation from placebo-treated ewes studied under the same protocol were also included. Blood-brain barrier function was quantified with the blood-to-brain transfer constant (K(i)) to alpha-aminoisobutyric acid. K(i) values were lower in cerebral cortex, caudate nucleus, hippocampus, superior colliculus, thalamus, medulla, and cervical spinal cord in fetuses of dexamethasone- than placebo-treated ewes at 60% but not 90% of gestation. Regional brain K(i) values demonstrated inverse correlations with increases in gestation and plasma cortisol concentrations in most brain regions. We conclude that maternal treatment with exogenous corticosteroids was associated with decreases in blood-brain barrier permeability at 60% but not 90% of gestation and that increases in gestation and endogenous cortisol concentrations were associated with ontogenic decreases in barrier permeability during fetal development.
Structure and Ligand Binding Determinants of the Recombinant Kringle 5 Domain of Human Plasminogen, (1998)

The X-ray crystal structure of the recombinant (r) kringle 5 domain of human plasminogen (K5HPg) has been solved by molecular replacement methods using K1HPg as a model and refined at 1.7 A resolution to an R factor of 16.6%. The asymmetric unit of K5HPg is composed of two molecules related by a noncrystallographic 2-fold rotation axis approximately parallel to the z-direction. The lysine binding site (LBS) is defined by the regions His33-Thr37, Pro54-Val58, Pro61-Tyr64, and Leu71-Tyr74 and is occupied in the apo-form by water molecules. A unique feature of the LBS of apo-K5HPg is the substitution by Leu71 for the basic amino acid, arginine, that in other kringle polypeptides forms the donor cationic center for the carboxylate group of omega-amino acid ligands. While wild-type (wt) r-K5HPg interacted weakly with these types of ligands, replacement by site-directed mutagenesis of Leu71 by arginine led to substantially increased affinity of the ligands for the LBS of K5HPg. As a result, binding of omega-amino acids to this mutant kringle (r-K5HPg[L71R]) was restored to levels displayed by the companion much stronger affinity HPg kringles, K1HPg and K4HPg. Correspondingly, alkylamine binding to r-K5HPg[L71R] was considerably attenuated from that shown by wtr-K5HPg. Thus, employing a rational design strategy based on the crystal structure of K5HPg, successful remodeling of the LBS has been accomplished, and has resulted in the conversion of a weak ligand binding kringle to one that possesses an affinity for omega-amino acids that is similar to K1HPg and K4HPg.
Functional interactions between a glutamine synthetase promoter and MYB proteins (2004)

In Scots pine (Pinus sylvestris), ammonium assimilation is catalysed by glutamine synthetase (GS) [EC 6.3.1.2], which is encoded by two genes, PsGS1a and PsGS1b. PsGS1b is expressed in the vascular tissue throughout the plant body, where it is believed to play a role in recycling ammonium released by various facets of metabolism. The mechanisms that may underpin the transcriptional regulation of PsGS1b were explored. The PsGS1b promoter contains a region that is enriched in previously characterized cis-acting elements, known as AC elements. Pine nuclear proteins bound these AC element-rich regions in a tissue-specific manner. As previous experiments had shown that R2R3-MYB transcription factors could interact with AC elements, the capacity of the AC elements in the PsGS1b promoter to interact with MYB proteins was examined. Two MYB proteins from loblolly pine (Pinus taeda), PtMYB1 and PtMYB4, bound to the PsGS1b promoter were able to activate transcription from this promoter in yeast, arabidopsis and pine cells. Immunolocalization experiments revealed that the two MYB proteins were most abundant in cells previously shown to accumulate PsGS1b transcripts. Immunoprecipitation analysis and supershift electrophoretic mobility shift assays implicated these same two proteins in the formation of complexes between pine nuclear extracts and the PsGS1b promoter. Given that these MYB proteins were previously shown to have the capacity to activate gene expression related to lignin biosynthesis, we hypothesize that they may function to co-regulate lignification, a process that places significant demands on nitrogen recycling, and GS, the major enzyme involved in the nitrogen recycling pathway.
Efficient targeted integration at leu1-32 and ura4-294 in Schizosaccharomyces pombe. (1994)

Homologous integration into the fission yeast Schizosaccharomyces pombe has not been well characterized. In this study, we have examined integration of plasmids carrying the leu1+ and ura4+ genes into their chromosomal loci. Genomic DNA blot analysis demonstrated that the majority of transformants have one or more copies of the plasmid vector integrated via homologous recombination with a much smaller fraction of gene conversion to leu1+ or ura4+. Non-homologous recombination events were not observed for either gene. We describe the construction of generally useful leu1+ and ura4+ plasmids for targeted integration at the leu1-32 and ura4-294 loci of S. pombe.
Mitochondrial Regulation of Caspase Activation by Cytochrome Oxidase and Tetramethylphenylenediamine via Cytosolic Cytochrome c Redox State (2007)

Cytochrome c release from mitochondria induces caspase activation in cytosols; however, it is unclear whether the redox state of cytosolic cytochrome c can regulate caspase activation. By using cytosol isolated from mammalian cells, we find that oxidation of cytochrome c by added cytochrome oxidase stimulates caspase activation, whereas reduction of cytochrome c by added tetramethylphenylenediamine (TMPD) or yeast lactate dehydrogenase/cytochrome c reductase blocks caspase activation. Scrape-loading of cells with this reductase inhibited caspase activation induced by staurosporine. Similarly, incubating intact cells with ascorbate plus TMPD to reduce intracellular cytochrome c strongly inhibited staurosporine-induced cell death, apoptosis, and caspase activation but not cytochrome c release, indicating that cytochrome c redox state can regulate caspase activation. In homogenates from healthy cells cytochrome c was rapidly reduced, whereas in homogenates from apoptotic cells added cytochrome c was rapidly oxidized by some endogenous process. This oxidation was prevented if mitochondria were removed from the homogenate or if cytochrome oxidase was inhibited by azide. This suggests that permeabilization of the outer mitochondrial membrane during apoptosis functions not just to release cytochrome c but also to maintain it oxidized via cytochrome oxidase, thus maximizing caspase activation. However, this activation can be blocked by adding TMPD, which may have some therapeutic potential.
Accuracy of non-invasive prenatal testing using cell-free DNA for detection of Down, Edwards and Patau syndromes: a systematic review and meta-analysis (2016)

CRD42014014947.
The continuing debate on self-monitoring of blood glucose in diabetes (2005)

April 2004 saw the publication of a much-needed consensus statement (Owens et al, 2004) on the use of self-monitoring of blood glucose (SMBG) by people with diabetes. Since the publication of the original document, the authors have attempted to obtain a wider consultation by involving healthcare professionals across the UK. This article aims to describe the measures taken to widen the debate in an attempt to provide as clear statements as possible in support of appropriate use of SMBG in people with diabetes.
Electric-Field Dependent Conformations of Single DNA Molecules on a Model Biosensor Surface (2012)

Despite the variety of nucleic acid sensors developed, we still do not have definite answers to some questions that are important to the molecular binding and, ultimately, the sensitivity and reliability of the sensors. How do the DNA probes distribute on the surface at the nanoscale? As the functionalized surfaces are highly heterogeneous, how are the conformations affected when the probe molecules interact with defects? How do DNA molecules respond to electric fields on the surface, which are applied in a variety of detection methods? With in situ electrochemical atomic force microscopy and careful tailoring of nanoscale surface interactions, we are able to observe the nanoscale conformations of individual DNA molecules on a model biosensor surface: thiolated DNA on a gold surface passivated with a hydroxyl-terminated alkanethiol self-assembled monolayer. We find that under applied electric fields, the conformations are highly sensitive to the choice of the alkanethiol molecule. Depending on the monolayer and the nature of the defects, the DNA molecules may either adopt a highly linear or a highly curved conformation. These unusual structures are difficult to observe through existing "ensemble" characterizations of nucleic acid sensors. These findings provide a step toward correlating target-binding affinity, selectivity, and kinetics to the nanoscale chemical structure of and around the probe molecules in practical nucleic acid devices.
Intra-articular glucocorticoid, bupivacaine and morphine reduces pain, inflammatory response and convalescence after arthroscopic meniscectomy (1998)

Convalescence after arthroscopic meniscectomy is dependent on pain and the inflammatory response. The aim of the study was therefore to investigate the effect of intra-articular bupivacaine + morphine + methylprednisolone versus bupivacaine + morphine or saline on postmeniscectomy pain, mobilisation and convalescence. In a double-blind randomized study 60 patients undergoing arthroscopic meniscectomy were allocated to intra-articular saline, intra-articular bupivacaine 150 mg + morphine 4 mg or the same dose of bupivacaine + morphine + intra-articular methylprednisolone 40 mg. All patients were instructed to resume normal activities immediately after operation. Pain during movement and walking, leg muscle force and joint effusion, use of crutches and duration of sick leave were assessed. Combined bupivacaine and morphine significantly reduced pain, time of immobilisation and duration of convalescence. Addition of methylprednisolone further reduced pain, use of additional analgesics, joint swelling and convalescence, improved muscle function and prevented the inflammatory response (acute phase protein) (P < 0.05). A multimodal analgesic and anti-inflammatory treatment may enhance post-arthroscopic convalescence, which depends on the trauma induced inflammatory response and pain.
Superoxide mediates the toxicity of paraquat for cultured mammalian cells. (1988)

To establish some of the necessary steps in the toxic action of paraquat for cultured mammalian cells, we isolated paraquat-resistant HeLa cells after lethal increments in concentration of paraquat in the medium. The paraquat-resistant cells had increased the cellular content of both the Mn-containing and the CuZn-containing superoxide dismutases. The effect of paraquat on the consumption of oxygen by the wild-type and the resistance cells was similar; in both cases exposure to paraquat for 24 or more hours produced similar proportions of cyanide-resistant consumption of oxygen, suggesting that paraquat entered both cells, underwent reduction, and donated electrons to molecular oxygen. When cultivated for 5 months in the absence of paraquat the paraquat-resistant cells maintained the increased cellular content of superoxide dismutases and remained resistant to paraquat. This observation suggested that resistance to paraquat might be caused by enrichment for the two superoxide dismutases and, further, that the increased cellular content of the two enzymes was not a response to growth under stressful conditions. NIH/3T3 cells whose content of superoxide dismutase was increased by transcription of the transfected cDNA for the human CuZn superoxide dismutase were also resistant to paraquat, suggesting strongly that paraquat promotes the formation of O2- as a necessary part of its cytotoxic effects in two types of cultured mammalian cells.
Controlled Hierarchical Architecture in Surface‐initiated Zwitterionic Polymer Brushes with Structurally Regulated Functionalities (2012)

Hierarchical polymer films with structurally regulated functionalities are achieved by integrating 2D and 3D structures to enable ultralow nonspecific protein binding and high loading of molecular recognition elements, such as antibodies.
Experimental Evaluation of a Nested Microphone Array With Adaptive Noise Cancellers (2004)

This paper proposes a near-field broadband adaptive beamforming scheme for intelligent computer telephony and teleconferencing applications, namely the nested microphone array with adaptive noise canceller (NMA-ANC). The NMA-ANC scheme incorporates an harmonically nested array with a nonuniformly subbanded multirate filter bank. Each subband array employs several near-field delay-filter-and-sum beamformers and an adaptive noise canceller (ANC). The proposed NMA-ANC is evaluated via a noise rejection experiment and dereverberation experiment performed in an anechoic chamber and a real conference room, respectively. The experiment data are recorded by a multichannel digital recording system developed using commercial off-the-shelf (COTS) equipments. A perceptual analysis/measurement system (PAMS) test is also carried out using a COTS digital speech level analyzer. The results of the experimental evaluation and PAMS test show that the proposed NMA-ANC scheme is able to improve the sound quality by adaptively rejecting multiple interfering signals and attenuating the reverberant noises and avoiding the desired signal cancellation.
New synthesis of aryl-substituted sulfonium salts and their applications in photoinitiated cationic polymerization (1990)

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTNew synthesis of aryl-substituted sulfonium salts and their applications in photoinitiated cationic polymerizationS. R. Akhtar, J. V. Crivello, J. L. Lee, and M. L. SchmittCite this: Chem. Mater. 1990, 2, 6, 732–737Publication Date (Print):November 1, 1990Publication History Published online1 May 2002Published inissue 1 November 1990https://pubs.acs.org/doi/10.1021/cm00012a027https://doi.org/10.1021/cm00012a027research-articleACS PublicationsRequest reuse permissionsArticle Views585Altmetric-Citations43LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InRedditEmail Other access optionsGet e-Alertsclose Get e-Alerts
Effects of Glucagon-Like Peptide 1 on the Kinetics of Glycogen Synthase a in Hepatocytes from Normal and Diabetic Rats* (1998)

Glucagon-like peptide 1(7-36)amide (GLP-1) is currently under investigation as a possible tool in the treatment of non-insulin-dependent diabetes mellitus. In addition to enhancing nutrient-stimulated insulin release, the peptide also favors glycogen synthesis and glucose use in liver, muscle, and adipose tissue. GLP-1 also activates glycogen synthase a in hepatocytes from both normal and diabetic rats. In the present study, the kinetic aspects of such an activation were investigated in hepatocytes from normal rats and from animals rendered diabetic induced by injection of streptozotocin, either in the adult age (insulin-dependent diabetes mellitus model) or in days 1 or 5 after birth (non-insulin-dependent diabetes mellitus models). GLP-1 increased, in a dose-dependent manner, glycogen synthase a activity in the hepatocytes from all groups studied. The activation of the enzyme reached a steady state within 1 min exposure to GLP-1, which, at 10(-12) M, caused a half-maximal activation. When comparing fed vs. overnight-starved normal rats, a somewhat lower basal activity of glycogen synthase a in fasted animals (P < 0.05) coincided with a greater relative increment in reaction velocity in response to GLP-1. The basal activity of glycogen synthase a and the relative extent of its inhibition by glucagon or activation by insulin and GLP-1 were modulated by the extracellular concentration of D-glucose. The activation of glycogen synthase a by either insulin or GLP-1 resulted not solely in an increase in maximal velocity but also in a decrease in affinity of the enzyme for uridine diphosphate-glucose; in diabetic animals, the capacity of insulin or GLP-1 to increase the maximal velocity and Michaelis-Menten constant were less marked than in normal rats. In conclusion, this study indicates that the GLP-1-induced activation of glycogen synthase a displays attributes of rapidity, sensitivity, and nutritional dependency that are well suited for both participation in the physiological regulation of enzyme activity and therapeutic purpose.
Transplantation of human islets without immunosuppression (2013)

Transplantation of pancreatic islets is emerging as a successful treatment for type-1 diabetes. Its current stringent restriction to patients with critical metabolic lability is justified by the long-term need for immunosuppression and a persistent shortage of donor organs. We developed an oxygenated chamber system composed of immune-isolating alginate and polymembrane covers that allows for survival and function of islets without immunosuppression. A patient with type-1 diabetes received a transplanted chamber and was followed for 10 mo. Persistent graft function in this chamber system was demonstrated, with regulated insulin secretion and preservation of islet morphology and function without any immunosuppressive therapy. This approach may allow for future widespread application of cell-based therapies.
Sites of single-strand breaks in DNA treated with a diol-epoxide of benz[a]anthracene (1983)

Evidence presented here indicates that r-8,t-9-dihydroxy-t-10,11-oxy-8,9,10,11-tetrahydrobenz[a]anthracene can introduce single-strand breaks into DNA. The positions of the single-strand breaks were determined in experiments in which the diol-epoxide was incubated with a defined 85 base pair DNA molecule that was radiolabelled at one 3' end with 32P. Analysis of the radiolabelled DNA fragments produced in these experiments in DNA sequencing gels showed that the breaks occur at most but not all guanines and at some adenines.
[Study of transcription and cleavage in vitro of HDV with HBV-specific hammerhead ribozyme]. (1999)

Recombinant Delt virus could serve as a vector for the delivery of a ribozyme specific for hepatitis B virus cleavage. Our data demonstrate the value of recombinant ribozyme as potential therapeutic agents for treatment of HBV infection. Further study about cleavage in vitro and in vivo will continue.