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Metal/semiconductor interactions affect electron transfer rates and this is central to photocatalytic hydrogen ion reduction. While this interaction has been studied in great detail on metal oxide semiconductors, not much is known of Au particles on top of polymeric semiconductors. The effects of gold nanoparticle size and dispersion on top of g-C3N4 were studied by core and valence level spectroscopy and transmission electron microscopy in addition to catalytic tests. The as-prepared, non-calcined catalysts displayed Au particles with uniform dimension (mean particle size = 1.8 nm) and multiple electronic states: XPS Au 4f7/2 lines at 84.9 and 87.1 eV (each with a spin-orbit splitting of 3.6-3.7 eV). These particles, which did not show localized surface plasmon resonance (LSPR), before the reaction, doubled in size after the reaction giving a pronounced LSPR at about 550 nm. The effect of the heating environment on these particles (in air or in H2) was further investigated. While heating in H2 gave Au nanoparticles of different shapes, heating under O2 gave exclusively spherical particles. Similar activity towards photocatalytic hydrogen ion reduction under UV excitation was seen in both cases, however. XPS Au 4f analyses indicated that an increase in deposition time, during catalyst preparation, resulted in an increase in the initial fraction of oxidized gold particles, which were easily reduced under hydrogen. The valence band region for Au/gC3N4 was further studied in an effort to compare it to what is already known for Au/metal oxide semiconductors. A shift of over 2 eV for the Au 5d doublets was noticed between reduced and oxidized gold particles with mean particle sizes between 2 and 6 nm, which is consistent with the final state effect. A narrow range of gold loading for optimal catalytic performance was seen, where it seems that a density of one Au particle per 10 × 10 nm2 is the most suitable. Particle size and shape had a minor effect on performance, which may indicate the absence of a plasmonic effect on the reaction rate.
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INTRODUCTION: In pediatric neurourology, clean intermittent catheterization (CIC) setting, and then self catheterization learning are important steps for children with neurogenic bladder. There is no adherence and satisfaction evaluation scale for children who are using self or hetero CIC. The aim of this article is to study the feasibility of using InCaSaQ (Intermittent Catheterization Satisfaction Questionnaire) and ICAS (Intermittent Catheterization Adherence Scale) in children, and to validate the first steps. PATIENTS AND METHODS: Scale validation monocentric study. Inclusion criterias were patients with neurogenic bladder, under the age of 18, using CIC (auto or hetero). The questionnaires ICAS and InCaSaQ were sent twice between 2017 March and April, and then filled by the child or his parents depending on who was doing the CIC. The internal concistency (Cronbach's alpha) measured the construct validity. The reproductibility was measured by the intraclass correlation cÅfficient (ICC) and the Wilcoxon and McNemar tests. Filling facility was evaluated for each score (evaluation with a 0 to 10 scale from the person who filled the questionnaire). RESULTS: Twenty two patients were included, and 50 questionnaires filled (25 ICAS and 25 InCaSaQ), twice each. Internal consistency was good for InCaSaQ (Cronbach's alpha>0,7) and so was ICAS and InCaSaQ reproductibility (ICC>0,7 for most of the questions). Patients under hetero-CIC had a worse adherence than parents of children under hetero-CIC (ICAS 3,25 versus 0,7 for children under hetero-CIC). The lowest InCaSaQ item was the way of throwing away their catheter. CONCLUSIONS: ICAS and InCaSaQ are interesting tools which can be used for children under auto and hetero-CIC. Studies with more patients will be necessary for finalizing the validation of these scales in the pediatric population. LEVEL OF PROOF: 4.
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Cateterismo Uretral Intermitente/métodos , Cooperación del Paciente , Satisfacción del Paciente , Vejiga Urinaria Neurogénica/terapia , Adolescente , Niño , Preescolar , Estudios de Factibilidad , Femenino , Humanos , Lactante , Masculino , Reproducibilidad de los Resultados , Autocuidado/métodos , Encuestas y CuestionariosRESUMEN
Loading of a co-catalyst on the surface of a semiconductor photocatalyst is often carried out without considering the effect of the loading procedure on the final product. The present study looks in detail at the effect that the loading method has on the morphology and final composition of platinum-based nanoparticles by means of XPS and TEM analysis. Additionally, reduction pre-treatments are performed to investigate how the coverage, crystallinity and composition of the NPs affect the photocatalytic H2 evolution. The activity of Pt-g-C3N4 can significantly be enhanced by controlling the properties of the co-catalyst NPs.
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STATEMENT OF FINDINGS: An inception cohort of 238 patients having peripheral joint synovitis of less than 12 months duration was evaluated clinically and followed prospectively for 1 year to determine the clinical significance of a number of rheumatoid arthritis (RA) associated autoantibodies. Serum samples collected at the time of the initial evaluation were tested for rheumatoid factor (RF) and antibodies to Sa (anti-Sa), RA-33, (pro)filaggrin [antifilaggrin antibody (AFA)], cyclic citrullinated peptide (anti-CCP), calpastatin, and keratin [antikeratin antibody (AKA)]. RF had a sensitivity of 66% and a specificity of 87% for RA. Anti-Sa, AFA, and anti-CCP all had a specificity of more than 90%, but a sensitivity of less than 50% for this diagnosis. Overall, there was a high degree of correlation between AFA, AKA, anti-Sa or anti-CCP, this being highest between anti-Sa and anti-CCP (odds ratio, 13.3; P < 0.001). Of the 101 patients who were positive for at least one of these four autoantibodies, 57% were positive for only one. Finally, anti-SA identified a subset of predominantly male RA patients with severe, erosive disease. Anti-SA, AFA and anti-CCP are all specific for early RA but, overall, have little additional diagnostic value over RF alone. Although these antibodies may preferentially recognize citrullinated antigens, the modest degree of concordance between them in individual patient sera suggests that it is unlikely a single antigen is involved in generating these responses.
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Artritis Reumatoide/diagnóstico , Artritis Reumatoide/inmunología , Autoanticuerpos/sangre , Sinovitis/diagnóstico , Sinovitis/inmunología , Enfermedad Aguda , Adulto , Especificidad de Anticuerpos , Artritis Reumatoide/epidemiología , Proteínas de Unión al Calcio/inmunología , Citrulina/inmunología , Coenzima A Ligasas , Estudios de Cohortes , Epítopos/inmunología , Femenino , Proteínas Filagrina , Prueba de Histocompatibilidad , Humanos , Proteínas de Filamentos Intermediarios/inmunología , Queratinas/inmunología , Masculino , Persona de Mediana Edad , Péptidos Cíclicos/inmunología , Valor Predictivo de las Pruebas , Proteínas/inmunología , Factor Reumatoide/sangre , Estudios Seroepidemiológicos , Sinovitis/epidemiologíaRESUMEN
The Sa system is a recently described immune system that has a specificity and positive predictive value of nearly 100% for rheumatoid arthritis (RA) in Asia, Europe and the Americas. Its sensitivity of 30-40% suggests that it identifies a subset of RA patients. Anti-Sa antibodies are present from disease onset and are predictive of disease severity. The immune reactants are plentiful in the target tissue: antigen is present in the synovium, IgG antibody in the fluid. Immunologically, Sa is a hapten-carrier antigen in which vimentin is the carrier and citrulline is the hapten. The citrullination of vimentin is closely related to apoptosis, and citrullinated vimentin is extremely sensitive to digestion by the ubiquitous calpains. Nevertheless, Sa is found in only a few cell lines. Calpastatin, the natural specific inhibitor of calpains, is also a RA-associated, albeit non-specific, autoimmune system. Is it possible that calpain-related apoptotic pathways could be prominent in cells containing Sa? The task is to reconcile the specificity of Sa/citrullinated proteins in a multifactorial and polygenic disease such as RA.
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Antígenos/inmunología , Artritis Reumatoide/inmunología , Haptenos/inmunología , HumanosRESUMEN
In Wegener's granulomatosis (WG), when the endogenous Proteinase 3 (PR3) of myeloid cells is translocated to the cell surface, a pathologically consequent interaction is believed to occur with classic anti-neutrophil cytoplasmic antibody (cANCA). In contrast, the exact origin of surface PR3 on cells of nonmyeloid origin is still debated. By various methods, PR3 mRNA and protein are easily demonstrated in myeloid cells but not in nonmyeloid cells. Exceptionally, the endothelial ECV304 cell line spontaneously produced PR3 mRNA but no PR3 protein. In the other nonmyeloid cells, we could not show cell surface PR3 either spontaneously or after TNFalpha stimulation. On the other hand, under serum-free conditions and using [(3)H]DFP-labeled HL-60 extract, a rapid, dose-dependent, saturable binding was demonstrated to both myeloid and nonmyeloid cells. That was reproduced with purified [(3)H]DFP-PR3. While we could not demonstrate cell surface PR3 on nonmyeloid cells after incubation with serum-containing supernatants of HL-60 cell cultures, we could do so after an overnight coculture period with HL-60 cell suspensions under the usual serum-containing culture conditions. Overall, our data would suggest that in vivo, the surface PR3 found on nonmyeloid cells is not endogenous but results from adsorption of PR3 extruded in their microenvironment by neighboring myeloid cells coming in close contact with them.
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Endotelio Vascular/citología , Serina Endopeptidasas/genética , Animales , Anticuerpos Anticitoplasma de Neutrófilos/inmunología , Western Blotting , Endotelio Vascular/enzimología , Células HL-60 , Humanos , Mieloblastina , Células Mieloides/enzimología , ARN Mensajero/metabolismo , Serina Endopeptidasas/sangreRESUMEN
OBJECTIVE: When polymorphonuclear neutrophils (PMN) and peripheral blood monocytes (PBMC) are stimulated with tumor necrosis factor alpha (TNF-alpha), preexisting granule stored proteinase 3 (PR3) is translocated to the surface of their plasma membrane. We investigated whether PR3 gene reactivation and new PR3 protein production were also features of priming by cytokine. METHODS: Normal human PMN and PBMC were isolated and stimulated in vitro with TNF-alpha. They were harvested at different intervals and subjected to total RNA and protein analysis. PR3 mRNA was identified by reverse transcription polymerase chain reaction, Northern blot, and sequencing. De novo PR3 synthesis was evaluated by metabolic labeling with [35S] methionine followed by immunoprecipitation using anti-neutrophil cytoplasmic antibodies from serum of patients with active Wegener's granulomatosis and mouse monoclonal anti-native PR3 antibodies. RESULTS: Resting PMN and PBMC do not express PR3 mRNA. During priming, PR3 mRNA appears in PMN at 2 h, peaks at 6 h, and has disappeared at 12 h. By comparison, in primed PBMC, PR3 mRNA appears at 6 h, peaks at 12 h, and disappears at 24 h. Immunoprecipitation of metabolically labeled PR3 revealed new synthesis of PR3 by both cell types, a process that was inhibited by cycloheximide. CONCLUSION: Primed PMN and PBMC can express PR3 mRNA and synthesize new PR3 protein, providing an alternative source to membrane PR3. Whether that small amount of inducible PR3 has a primary structure, a localization, or a role different from those of preformed PR3 stored in granules remains to be clarified.
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Autoantígenos/biosíntesis , Granulomatosis con Poliangitis/sangre , Leucocitos Mononucleares/enzimología , Neutrófilos/enzimología , Serina Endopeptidasas/biosíntesis , Autoantígenos/genética , Northern Blotting , Células HL-60/enzimología , Células HeLa/enzimología , Humanos , Activación de Linfocitos , Mieloblastina , ARN/análisis , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Serina Endopeptidasas/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
The recent discovery of cyclooxygenase-2 (COX-2), an isoenzyme associated mainly with inflammation created the need to reevaluate cyclooxygenase inhibitors with reliable screening methods. In the present study we standardized a technique to determine the IC50S of cyclooxygenase inhibitors on recombinant human COX-1 and COX-2 expressed in mammalian cells and used it to study the compounds tenoxicam, aspirin and indomethacin. The IC50S of aspirin, indomethacin and tenoxicam for human COX-1 were 0.41 +/- 0.07 microgram/ml, 0.008 +/- 0.003 microgram/ml, and 7.94 +/- 3.28 micrograms/ml, respectively, and for human COX-20.64 +/- 0.16 microgram/ml, 0.09 +/- 0.05 microgram/ml, and 10.61 +/- 1.50 micrograms/ml, for aspirin, indomethacin, and tenoxicam. Tenoxicam had the lowest IC50hCOX-2/IC50hCOX-1 ratio (1.34), followed by aspirin (1.53) and indomethacin (10.82). The system described in the present study provides a simple and efficient way to determine the specificity of NSAID inhibition for each of the human cyclooxygenase isoenzymes separately.
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Aspirina/farmacología , Inhibidores de la Ciclooxigenasa/farmacología , Indometacina/farmacología , Isoenzimas/efectos de los fármacos , Piroxicam/análogos & derivados , Prostaglandina-Endoperóxido Sintasas/efectos de los fármacos , Animales , Células COS , Ciclooxigenasa 1 , Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa 2 , Relación Dosis-Respuesta a Droga , Humanos , Isoenzimas/genética , Proteínas de la Membrana , Piroxicam/farmacología , Prostaglandina-Endoperóxido Sintasas/genética , Proteínas Recombinantes/efectos de los fármacos , TransfecciónAsunto(s)
Southern Blotting/métodos , ADN/análisis , Bacteriófago lambda/química , ADN Viral/análisis , ADN Viral/metabolismo , Desoxirribonucleasa EcoRI/metabolismo , Desoxirribonucleasa HindIII/metabolismo , Electroforesis en Gel de Agar/métodos , Etanol/farmacología , Hibridación de Ácido NucleicoRESUMEN
Ro and La RNP complexes were reassembled from in vitro labelled hY5 RNA and HeLa cell extracts. These complexes were then visualized through retardation of migration of labelled hY5 RNA in non-denaturing polyacrylamide gels. Three major complexes (named A, B, and C) were formed when crude cellular extracts (S100 fraction) were used. Using monospecific anti-60-kD Ro (Ro60) and anti-La antibodies to retard RNPs containing these antigens during migration in the gels, the three major complexes were shown to contain Ro60 (C), La (B), or both proteins (A). The specificity of RNA-protein interactions in the reassembled complexes was further demonstrated using two 3'-shortened hY5 RNA transcripts lacking the La-binding site (hY5-Alu I RNA) and both the Ro60 and La-binding sites (hY5-Hha I RNA). hY5-Hha I RNA still formed a single, minor complex when incubated with S100 extract, suggesting interaction with a yet undefined protein. In addition, we used the capacity of specific antibodies to retard the migration of the reassembled complexes to design a detection assay for anti-Ro and anti-La autoantibodies. Using 84 human sera, our assay was shown to approximate the specificity and sensitivity of an immunoprecipitation assay where 32P-labelled cell extracts are used as source of antigens. Our assay may be used to detect low levels of antibodies to conformational determinants on Ro60 and La proteins in human sera and antibody preparations.
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Autoantígenos/metabolismo , ARN Citoplasmático Pequeño , Ribonucleoproteínas/metabolismo , Electroforesis en Gel de Poliacrilamida , Humanos , Microesferas , Unión Proteica/genética , ARN/análisis , Antígeno SS-BRESUMEN
We recently reported the identification in human anti-Ro serum of Abs specifically immunoprecipitating deproteinized hY5 RNA. In the present report, we characterized the epitopes recognized by anti-hY5 RNA Abs. Using deletion and site-directed mutagenesis of hY5 cDNA and in vitro transcribed RNAs with intact and 3'-shortened ends, we have defined two conformational antigenic determinants distinct from the regions known to bind Ro and La proteins. One of these epitopes (epitope A) is present in the middle portion of hY5 RNA and is dependent on the presence of a four-nucleotide sequence (AACC at position 58-61) that may form a single-stranded loop. Deleting these four nucleotides or modifying the stem structures proximal or distal to this loop abolishes recognition of the mutated RNAs by Abs. The second epitope (epitope B) requires the presence of another four-nucleotide sequence (CUUG at position 74-77) in between the Ro and La binding sites. Deleting this CUUG sequence or modifying nucleotides on the 5' side of the stem structure below the Ro60 binding site severely compromises the interaction with Abs. Since Abs to deproteinized hY RNAs are restricted to anti-hY5 RNA and target determinants not involved in interactions with known hY5 RNA-binding proteins, human RohY5 particles may play a direct role in the immunization process, leading to the production of anti-hY5 RNA autoantibodies.
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Autoantígenos/inmunología , Epítopos Inmunodominantes/inmunología , ARN Citoplasmático Pequeño , ARN/inmunología , Ribonucleoproteínas/inmunología , Especificidad de Anticuerpos , Autoanticuerpos/inmunología , Secuencia de Bases , Electroforesis en Gel de Poliacrilamida , Humanos , Epítopos Inmunodominantes/química , Datos de Secuencia Molecular , ARN/química , Antígeno SS-BRESUMEN
This study evaluated physicians' use of the occurrence of tinnitus as a tool to establish the optimal dosage of salsalate, a nonacetylated salicylate, in patients with arthritis treated in routine clinical practice. The use of printed educational materials to improve compliance was also studied prospectively. A total of 782 patients were enrolled in this 3-week study by 95 general practitioners in an office setting. Of the 771 assessable patients, 90.0% had osteoarthritis, 9.7% had rheumatoid arthritis, and 0.3% had both types of arthritis. Most patients experienced improvement of symptoms after 3 weeks of treatment. There were no differences in the rates of improvement at the first and third weeks of treatment between patients with osteoarthritis and patients with rheumatoid arthritis. In addition, duration of arthritis had no effect on rates of improvement. Rates of patient satisfaction tended to increase over the study period. Rates of patient satisfaction did not differ significantly at the first and third weeks between patients who did not receive printed educational materials and whose who did not. Treatment was discontinued in 234 patients (30.4%) because of side effects. The most frequent reasons for discontinuation were gastrointestinal symptoms (n = 102; 13.2%) and tinnitus (n = 52; 6.7%). The clinical effectiveness and safety of salsalate were confirmed in patients with arthritis in routine clinical practice settings.
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Artritis Reumatoide/tratamiento farmacológico , Osteoartritis/tratamiento farmacológico , Salicilatos/uso terapéutico , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Satisfacción del Paciente , Estudios ProspectivosRESUMEN
OBJECTIVE: To study the association of maternal antibodies to Ro(SSA) and/or La(SSB) with isolated complete congenital heart block (CCHB) in children according to the child's age at detection. METHODS: Sera from 17 mothers of 18 children with CCHB of unidentified cause were studied. Autoantibodies were measured by double immunodiffusion, enzyme linked immunosorbent assay (ELISA), Western blot, and immunoprecipitation from cell extracts. Statistical analysis used the chi 2 test with Yates' correction. RESULTS: CCHB was diagnosed in 12 children of 11 mothers before the age of 3 mo (Group A) and in 6 children of 6 mothers after the age of 17 mo (Group B). Seven Group A mothers and no Group B mother had connective tissue disorders; autoantibodies were found in 9/11 Group A and in 1/6 Group B mothers (p < 0.01). Eight Group A children needed a pacemaker and one other died of cardiac insufficiency, whereas only one of the 6 Group B children needed a pacemaker. Interestingly, this latter child was the only one from Group B whose mother's serum contained autoantibodies. Irrespective of their age at diagnosis, the children with CCHB who needed a pacemaker and the one who died were born to mothers with autoantibodies (p < 0.001). CONCLUSION: CCHB detected before the age of 3 mo is highly associated with the presence of anti-Ro(SSA)/La(SSB) in the mothers, while CCHB diagnosed later is generally not. For epidemiological studies, the former type should be considered early onset as opposed to late onset CCHB in the latter type. Establishing this clinicoserological distinction is also important for the children, since it alerts the clinician to a more severe prognosis (necessity of a pacemaker), even in the rare occurrence of late diagnosed CCHB.
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Autoanticuerpos/sangre , Autoantígenos/inmunología , Bloqueo Cardíaco/diagnóstico , ARN Citoplasmático Pequeño , Ribonucleoproteínas/inmunología , Adolescente , Western Blotting , Niño , Preescolar , Ensayo de Inmunoadsorción Enzimática , Femenino , Bloqueo Cardíaco/congénito , Bloqueo Cardíaco/inmunología , Humanos , Inmunodifusión , Lactante , Masculino , Pruebas de Precipitina , Pronóstico , Antígeno SS-BRESUMEN
Immunoprecipitation (IP) of radiolabeled PMN extracts was used as the gold standard for anti-proteinase 3 (PR-3) autoantibody detection to validate immunofluorescence (IF) and alpha granule (alpha) ELISA. A myeloperoxidase (MPO) ELISA was also used in parallel. We studied 48 patients with strictly defined vasculitic syndromes in the initial active phase of their disease. The 3 methods confirmed the high (> 90%) sensitivity and specificity of anti-PR-3 for patients with Wegener's granulomatosis (WG). Similarly, a high (86%) sensitivity of MPO-ELISA was found in microscopic polyarteritis as defined. Using alpha-ELISA, we could not improve the detection rate of anti-PR-3 obtained by IF-cANCA-pattern reading. Moreover, a small proportion (< 15%) of biopsy-proven WG patients had anti-MPO antibodies detected by IF, usually as pANCA but also, even if rarely, as bona fide cANCA (< 5%). Thus, IF would seem to be the most reliable screening method and alpha-ELISA should be used for confirmation. On the other hand, because MPO-ELISA detected twice as many anti-MPO positive sera as did pANCA pattern reading by IF, we suggest that in the clinical context of a vasculitis, MPO-ELISA should also be used as a screening test. Although IP is not designed for routine clinical use, it should be required when reporting the presence of anti-PR-3 in vasculitis-like diseases that are fertile grounds for false positive reactions.
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Autoanticuerpos/análisis , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Neutrófilos/inmunología , Pruebas de Precipitina , Vasculitis/diagnóstico , Adulto , Anciano , Anticuerpos Anticitoplasma de Neutrófilos , Autoanticuerpos/inmunología , Femenino , Granulomatosis con Poliangitis/diagnóstico , Granulomatosis con Poliangitis/inmunología , Humanos , Isoflurofato , Masculino , Persona de Mediana Edad , Poliarteritis Nudosa/diagnóstico , Poliarteritis Nudosa/inmunología , Sensibilidad y EspecificidadRESUMEN
Anti-Ro sera immunoprecipitate Ro ribonucleoproteins (RNPs) from human cell extracts. Ro RNPs are biochemically heterogeneous particles whose functions are unknown and whose exact composition remains controversial. In addition to 60-kD Ro and to La proteins, a 52-kD polypeptide (p52) has been proposed to be a stable component of the Ro RNPs. To confirm the immunological studies supporting this hypothesis, we have biochemically purified Ro RNPs from HeLa cells using non-denaturing conditions. Ro RNPs segregated into three distinct populations, one of which only contained hY5 RNA (RohY5 RNPs). No p52 co-purified with Ro RNPs. Despite the absence of p52, purified Ro RNPs had biochemical and immunological properties identical to those of unfractionated Ro RNPs. Many anti-Ro sera only recognize p52 in immunoblots, and are said to be monospecific anti-p52. Preincubation with purified RohY5 RNPs (free of p52) of all human anti-Ro (including so-called monospecific anti-p52) sera abolished their capacity to immunoprecipitate Ro RNPs from unfractionated HeLa cell extracts. Conversely, preincubation of anti-Ro sera with purified p52 protein specifically inhibited recognition of p52 in immunoblots, but did not interfere with immunoprecipitation of Ro RNPs. Our data demonstrate that anti-p52 antibodies do not target intact Ro RNPs, nor do they target the native 60-kD Ro protein. Contrary to previous reports, p52 protein is not a stable component of antigenically intact Ro RNPs.