RESUMEN
Nitrous oxide (N2O) is a widely used anesthetic agent. We report two patients with sickle cell disease (SCD) who presented with complications following the use of N2O. Patient 1, a 15-year-old girl, presented severe hyperhomocysteinemia, pancytopenia, vitamin B12 deficiency, and peripheral polyneuropathy after massive use of N2O for pain management. At the 1-year follow-up, hyperhomocysteinemia and B12 deficiency had resolved, but she had persisting mild symptoms of polyneuropathy. Patient 2, a 17-year-old boy, presented only severe hyperhomocysteinemia, only partially corrected by initial B12 supplementation. Careful monitoring of N2O use, especially in patients with SCD, is mandatory to prevent complications.
Asunto(s)
Anemia de Células Falciformes/tratamiento farmacológico , Anestésicos por Inhalación/efectos adversos , Hiperhomocisteinemia/inducido químicamente , Óxido Nitroso/efectos adversos , Enfermedades del Sistema Nervioso Periférico/inducido químicamente , Adolescente , Anestésicos por Inhalación/uso terapéutico , Femenino , Humanos , Hiperhomocisteinemia/diagnóstico , Masculino , Nervio Mediano/efectos de los fármacos , Nervio Mediano/fisiopatología , Óxido Nitroso/uso terapéutico , Enfermedades del Sistema Nervioso Periférico/diagnóstico , Nervio Peroneo/efectos de los fármacos , Nervio Peroneo/fisiopatología , Índice de Severidad de la Enfermedad , Nervio Tibial/efectos de los fármacos , Nervio Tibial/fisiopatologíaRESUMEN
Botulism is an uncommon severe neuromuscular disorder. We report two recent cases of confirmed infant botulism diagnosed in an 11-week and a 5-month-old infant along with electroneuromyogram (ENMG) findings. Then, we discuss the EMG features of infant botulism. In severe forms of infant botulism, presence of these features might help decide to use botulinum immune globulin. To our knowledge, case 1 is the first case reported in France based on confirmed dust contamination.
RESUMEN
UNLABELLED: We demonstrate the clinical interest of bone texture analysis with a new high resolution X-ray device. We have found that the combination of BMD and texture parameter values provided a better assessment of the fracture risk than that obtainable solely by BMD measurement. INTRODUCTION: Osteoporosis is characterized by BMD and trabecular bone microarchitecture. We have developed a new high-resolution X-ray device with direct digitization. The aim of this study was to demonstrate in a multicenter case control study the clinical interest of bone texture analysis with this new device. METHODS: In this cross-sectional multicenter case-control population study in post-menopausal women, 159 osteoporotic fractures were compared with 219 control cases. Images were obtained on calcaneus with a direct digital X-ray device (BMA, D3A Medical Systems). Co-occurrence, run-length matrices and the fractal parameter Hmean were evaluated. BMD was measured at the lumbar spine (LS), femoral neck (FN) and total hip (TH) by DXA. RESULTS: The three texture parameters were significantly lower in osteoporotic fracture cases than in control cases. These differences persisted after adjustment for TH BMD. Receiver operating characteristic curves were used to compare the discriminant capacity of texture parameters and BMD measurements for fracture. The highest areas under curve (AUC) were 0.721 for TH BMD and 0.706 for Hmean (AUC THBMD vs. AUC Hmean, p = NS). We determined the threshold between high and low Hmean parameter values and then the odds ratios (OR) of fracture for low Hmean, for BMD < or =2.5 SD in the T-score and for combinations of both parameters. The OR of fracture for low H was 2.72 (95% CI, 1.36-5.4). For a FN BMD < or = -2.5 SD, the OR of 4.78 (2.19-10.43) shifted to 14.06 (4.41-44.85) adding H. CONCLUSIONS: These data confirmed the clinical interest of the combination of BMD and texture parameters to improve the assessment of the risk of fracture other that obtainable by the sole BMD measurement.
Asunto(s)
Densidad Ósea , Huesos/diagnóstico por imagen , Osteoporosis Posmenopáusica/complicaciones , Absorciometría de Fotón , Anciano , Anciano de 80 o más Años , Calcáneo/diagnóstico por imagen , Estudios de Casos y Controles , Estudios Transversales , Femenino , Cuello Femoral/diagnóstico por imagen , Fracturas Espontáneas/diagnóstico por imagen , Fracturas Espontáneas/patología , Cadera/diagnóstico por imagen , Humanos , Vértebras Lumbares/diagnóstico por imagen , Persona de Mediana Edad , Oportunidad Relativa , Osteoporosis Posmenopáusica/diagnóstico por imagen , Riesgo , UltrasonografíaRESUMEN
Identification of modified amino acids can be a challenging part for Edman degradation sequence analysis, largely because they are not included among the commonly used phenylthiohydantion amino acid standards. Yet many can have unique retention times and can be assigned by an experienced researcher or through the use of a guide showing their typical chromatography characteristics. The Edman Sequencing Research Group (ESRG) 2005 study is a continuation of the 2004 study, in which the participating laboratories were provided a synthetic peptide and asked to identify the modified amino acids present in the sequence. The study sample provided an opportunity to sequence a peptide containing a variety of modified amino acids and note their retention times relative to the common amino acids. It also allowed the ESRG to compile the chromatographic properties and intensities from multiple instruments and tabulate an average elution position for these modified amino acids on commonly used instruments. Participating laboratories were given 2000 pmoles of a synthetic peptide, 18 amino acids long, containing the following modified amino acids: dimethyl- and trimethyl-lysine, 3-methyl-histidine, N-carbamyl-lysine, cystine, N-methyl-alanine, and isoaspartic acid. The modified amino acids were interspersed with standard amino acids to help in the assessment of initial and repetitive yields. In addition to filling in an assignment sheet, which included retention times and peak areas, participants were asked to provide specific details about the parameters used for the sequencing run. References for some of the modified amino acid elution characteristics were provided and the participants had the option of viewing a list of the modified amino acids present in the peptide at the ESRG Web site. The ABRF ESRG 2005 sample is the seventeenth in a series of studies designed to aid laboratories in evaluating their abilities to obtain and interpret amino acid sequence data.
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Aminoácidos/análisis , Aminoácidos/química , Análisis de Secuencia de Proteína , Secuencia de Aminoácidos , Datos de Secuencia Molecular , Compuestos Organofosforados , Péptidos/química , Feniltiohidantoína/químicaRESUMEN
The two conventional tests to detect pyrogen contaminants in injectable pharmaceutical drugs are the Rabbit Model and the Limulus amoebocyte lysate (LAL) test. To replace these models, a new system on human whole blood is developed, using the release of Interleukin 1 beta (IL1beta) after cell stimulation with gram-positive and gram negative pyrogens. The purpose of this study was to validate the ENDOSAFE-IPT kit using the quantitative ELISA enzyme immunoassay. The assay is divided into two parts: blood cell stimulation with Lipopolysaccharides (LPS) and Lipoteichoic acid (LTA) and quantitation of IL1beta using the ELISA method. In each assay, blood from a particular donor were stimulated with the Endotoxin Standard, and with a sample of a commercial antibiotic preparation (Clavulanic acid/Ticarcillin) spiked with the Endotoxin Standard. LTA from Bacillus subtilis and a sample of diphtheria toxoid were also used. At least, six assays were tested. A polynomial regression of the Endotoxin Standard series showed a correlation coefficient greater than 0.99. The spiked antibiotic sample recoveries were 50-121%. The LTA quantitation limit was 0.1 microg/ml and the range of detection of pyrogens from Gram positive diphtheria toxoid was 0.77 to 2.5 EEU/ml. The IL1beta production varied markedly between donors. However the coefficient of variation was less than 20 % intra-assay. In conclusion, the ENDOSAFE-IPT kit can be used for the quantitative and qualitative detection of pyrogens from Gram negative and Gram positive bacteria.
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Contaminación de Medicamentos , Interleucina-1beta/metabolismo , Preparaciones Farmacéuticas/análisis , Pirógenos/análisis , Bacillus subtilis/patogenicidad , Biomarcadores/análisis , Biomarcadores/metabolismo , Sangre , Ácido Clavulánico , Toxoide Diftérico , Ensayo de Inmunoadsorción Enzimática , Bacterias Gramnegativas/patogenicidad , Bacterias Grampositivas/patogenicidad , Humanos , Interleucina-1beta/análisis , Lipopolisacáridos , Control de Calidad , Reproducibilidad de los Resultados , Ácidos Teicoicos , TicarcilinaRESUMEN
Peptide fragments of self-proteins bound to major histocompatibility complex molecules within the thymus are important for positively selecting T cell receptor (TCR)-bearing CD4(+)CD8(+) double positive (DP) thymocytes for further maturation. The relationship between naturally processed thymic self-peptides and TCR-specific cognate peptides is unknown. Here we employ HPLC purification of peptides released from H-2K(b) molecules of the C57BL/6 thymus in conjunction with mass spectrometry (MS) and functional profiling to identify a naturally processed K(b)-bound peptide positively selecting the N15 TCR specific for the vesicular stomatitis virus octapeptide (VSV8) bound to K(b). The selecting peptide was identified in 1 of 80 HPLC fractions and shown by tandem MS (MS/MS) sequencing to correspond to residues 68-75 of the MLRQ subunit of the widely expressed mitochondrial NADH ubiquinone oxidoreductase (NUbO(68-75)). Of note, the peptide differs at six of its eight residues from the cognate peptide VSV8 and functions as a weak agonist for mature CD8 single positive (SP) N15 T cells, with activity 10,000-fold less than VSV8. In N15 transgenic (tg) recombinase activating gene 2(-/)- transporter associated with antigen processing 1(-/)- fetal thymic organ culture, NUbO(68-75) induces phenotypic and functional differentiation of N15 TCR bearing CD8 SP thymocytes. Failure of NUbO(68-75) to support differentiation of a second K(b)-restricted TCR indicates that its inductive effects are not general.
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Presentación de Antígeno , Antígenos H-2/inmunología , Mitocondrias/inmunología , Oligopéptidos/inmunología , Receptores de Antígenos de Linfocitos T/inmunología , Timo/inmunología , Virus de la Estomatitis Vesicular Indiana/inmunología , Animales , Complejo I de Transporte de Electrón , Ratones , Ratones Endogámicos C57BL , Mitocondrias/enzimología , NADH NADPH Oxidorreductasas/inmunología , Oligopéptidos/aislamiento & purificación , Fragmentos de Péptidos/inmunología , Receptores de Antígenos de Linfocitos T/genética , Selección Genética , Timo/citologíaRESUMEN
In the course of characterizing proteins present in a preparation of vesicles from rat adipocytes containing glucose transporters, we examined a protein that migrated at 115 kDa upon SDS gel electrophoresis (designated vp115). Sequences of tryptic peptides were obtained, and from this information the cDNA for rat vp115 was cloned. The cDNA encodes an open reading frame for a protein of 121 kDa. Computer-aided sequence analysis predicted that vp115 has a potential membrane-inserted or membrane-spanning domain toward its amino terminus, followed by five C2 domains. Immunoblotting revealed that vp115 was not actually a component of the glucose transporter-containing vesicles, was most abundant in the plasma membranes and high density microsome fractions of rat adipocytes, and was expressed in all the major rat tissues.
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Adipocitos/metabolismo , Proteínas de la Membrana/genética , Adipocitos/química , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Immunoblotting , Proteínas de la Membrana/análisis , Proteínas de la Membrana/química , Datos de Secuencia Molecular , Ratas , Homología de Secuencia , Fracciones Subcelulares/metabolismoRESUMEN
In the course of our examination of proteins associated with the GLUT4-containing vesicles of rat adipocytes we have identified a new 22 kDa member of the family of endoplasmic reticulum (ER) proteins known as reticulons. The protein, which we refer to as vp20, was purified from a preparation of GLUT4-containing vesicles of rat adipocytes, and tryptic peptides were micro-sequenced. From this information a cDNA encoding a single open reading frame for a protein of 22 kDa was cloned. This protein is homologous to known members of the reticulon protein family. vp20 has two hydrophobic stretches of about 35 amino acids that could be membrane spanning domains and an ER retention motif at its carboxy-terminus. vp20 was most abundant in the high density microsome fraction of adipocytes, which is the fraction most enriched in ER. Only a small fraction of vp20 was present in the GLUT4 vesicle population, and that fraction appears to be due to ER vesicles that were non-specifically bound to the adsorbent. Analysis of tissue distribution of vp20 in rats revealed that it is concentrated in muscle, fat and the brain.
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Adipocitos/metabolismo , Proteínas de la Membrana/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Encéfalo/metabolismo , Clonación Molecular , ADN Complementario/química , Proteínas de la Membrana/química , Proteínas de la Membrana/aislamiento & purificación , Microsomas/metabolismo , Datos de Secuencia Molecular , Músculos/metabolismo , Proteínas de la Mielina , Proteínas Nogo , RatasRESUMEN
Copolymer 1 (Cop 1, poly (Y, E, A, K)) is a random synthetic amino acid copolymer effective in the treatment of relapsing forms of multiple sclerosis (MS). Cop 1 binds promiscuously, with high affinity and in a peptide-specific manner to purified MS-associated HLA-DR2 (DRB1*1501) and rheumatoid arthritis-associated HLA-DR1 (DRB1*0101) or HLA-DR4 (DRB1*0401) molecules. In the present work at least 95% of added Cop 1 could be bound to recombinant "empty" HLA-DR1 and -DR4, and 80% could be bound to HLA-DR2 proteins. Amino acid composition, HPLC profiles, and sequencing patterns of Cop 1 eluted by acid extraction from HLA-DR molecules were similar to those of the unseparated Cop 1. Protruding N-terminal ends of Cop 1 bound to HLA-DR1, -DR2, or -DR4 molecules were then treated with aminopeptidase I, followed by elution, HPLC, and pool sequencing. In contrast to untreated or unbound Cop 1, this material exhibited distinct motifs at some positions with increases in levels of E at the first and second cycles, of K at the second and third cycles, and of Y (presumably at P1 of the bound peptide) at the third to fifth cycles, regardless of the HLA-DR molecule employed. No preference was seen at the following cycles that were mainly A. These first pooled HLA-DR binding epitopes provide clues to the components of Cop 1 that are biologically active in suppressing MS and possibly rheumatoid arthritis.
Asunto(s)
Artritis Reumatoide/metabolismo , Antígenos HLA-DR/metabolismo , Esclerosis Múltiple/metabolismo , Fragmentos de Péptidos/inmunología , Fragmentos de Péptidos/metabolismo , Péptidos/inmunología , Péptidos/metabolismo , Polímeros/metabolismo , Proteínas de Saccharomyces cerevisiae , Secuencia de Aminoácidos , Aminoácidos/análisis , Aminopeptidasas/química , Anticuerpos/metabolismo , Artritis Reumatoide/inmunología , Sitios de Unión/inmunología , Sitios de Unión de Anticuerpos , Cromatografía Líquida de Alta Presión , Acetato de Glatiramer , Antígenos HLA-DR/inmunología , Antígeno HLA-DR1/metabolismo , Antígeno HLA-DR2/metabolismo , Antígeno HLA-DR4/metabolismo , Humanos , Esclerosis Múltiple/inmunología , Fragmentos de Péptidos/aislamiento & purificación , Péptidos/aislamiento & purificación , Polímeros/aislamiento & purificaciónRESUMEN
VP39 is a bifunctional mRNA-modifying protein that acts as both an mRNA cap-specific 2'-O-methyltransferase and a processivity factor for VP55, the vaccinia poly(A) polymerase catalytic subunit. Although regions of the protein surface required for methyltransferase function are well defined, it has been unclear whether the protein polyadenylylation function requires direct RNA contact and, if so, where the contact site(s) might be located on the protein surface. Here, we show that the VP55-VP39 heterodimer forms a stable complex with a 50mer oligonucleotide bearing a U2-N25-U motif, as opposed to the U2-N15-U motif that is optimal for stable complex formation with VP55 alone. An oligonucleotide bearing a U2-N25-U motif in which the downstream U residue is replaced with 4thioU can be efficiently photocrosslinked to VP39, but only in the context of the VP55-VP39 heterodimer. By partial proteolysis of end-labeled VP39, the site of oligonucleotide photocrosslinking was localized to the region of VP39 between residues Lys90 and Arg122. Peptide microsequencing and confirmatory mutagenesis identified the side-chain of Arg107 as the photocrosslinking site. Substitution of this residue with lysine abolished photocrosslinking entirely, consistent with the established RNA binding role of arginine in other RNA-binding proteins. This study provides clear evidence for a polyadenylylation-specific RNA-contact site on the surface of VP39, which is distinct from the RNA-binding methyltransferase "cleft" characterized in recent crystallographic and biochemical studies.
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ARN Mensajero/metabolismo , ARN Viral/metabolismo , Virus Vaccinia/metabolismo , Proteínas Virales/química , Proteínas Virales/metabolismo , Secuencia de Bases , Sitios de Unión , Reactivos de Enlaces Cruzados , Dimerización , Metiltransferasas/química , Metiltransferasas/metabolismo , Modelos Moleculares , Mutagénesis Sitio-Dirigida , Oligonucleótidos/química , Oligonucleótidos/genética , Polinucleotido Adenililtransferasa/química , Polinucleotido Adenililtransferasa/metabolismo , Unión Proteica , Conformación Proteica , Caperuzas de ARN/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Virus Vaccinia/genética , Proteínas Virales/genéticaAsunto(s)
Continuidad de la Atención al Paciente/organización & administración , Prestación Integrada de Atención de Salud/organización & administración , Reforma de la Atención de Salud/organización & administración , Atención Perinatal/organización & administración , Medicina Preventiva/organización & administración , Femenino , Humanos , EmbarazoRESUMEN
Steady-state captopril renography (SSCR) is an original technique for assessing the captopril-induced increase in technetium-99m mercaptoacetyltriglycine (99mTc-MAG3) mean parenchymal transit time (MPTT) in kidneys affected with functional renal artery stenosis (RAS). The steady-state parenchymal activity achieved by constant infusion of 99mTc-MAG3 is directly linked to the MPTT of the radiopharmaceutical. This steady-state parenchymal activity was continuously monitored from 15 min before to 60 min after a single dose of captopril in order to detect possible disruption of the steady state. SSCR was performed in 11 hypertensive patients with unilateral RAS and in two with RAS of a solitary kidney before renal revascularization (RR). In four of these patients, an additional SSCR was performed after RR. Of the ten patients whose hypertension was cured or improved by RR, one presented an uninterpretable SSCR and six presented a positive SSCR on the affected side. Control SSCR performed in four of these six patients was bilaterally negative. SSCR was also bilaterally negative in the three patients who showed no blood pressure response to RR. These preliminary results tend to indicate that, in spite of the stability of pre- and post-captopril hydration and data acquisition conditions allowed by this steady-state technique, the sensitivity is lower than expected. However, the reason for the false-negative results does not seem to be inherent to SSCR.
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Antihipertensivos , Captopril , Hipertensión Renovascular/diagnóstico por imagen , Renografía por Radioisótopo/métodos , Radiofármacos , Tecnecio Tc 99m Mertiatida , Adulto , Animales , Antihipertensivos/farmacología , Captopril/farmacología , Humanos , Hipertensión Renovascular/terapia , Persona de Mediana Edad , Monitoreo Fisiológico , Radiofármacos/farmacocinética , Sensibilidad y Especificidad , Tecnecio Tc 99m Mertiatida/farmacocinética , Factores de TiempoRESUMEN
The Tat protein is a transcriptional activator which is required for efficient human immunodeficiency virus 1 (HIV-1) gene expression Tat stimulates HIV-1 transcriptional elongation by increasing the processivity of RNA polymerase II. To address whether Tat-mediated effects on HIV-1 gene expression are due to modulation in the phosphorylation of the RNA polymerase II C-terminal domain (CTD), we developed a purification protocol to identify cellular kinases that are capable of binding to Tat and hyperphosphorylating the RNA polymerase II CTD. A 600 kDa protein complex with these properties was isolated, and specific components were identified using peptide microsequence analysis. This analysis indicated that proteins comprising the multi-subunit TFIIH complex, in addition to several novel factors, were associated with Tat using both in vitro and in vivo analysis. The Tat-associated kinase bound to the activation domain of Tat, and its ability to hyperphosphorylate RNA polymerase II was markedly stimulated by Tat. Furthermore, the addition of the Tat-associated kinase to in vitro transcription assays stimulated the ability of Tat to activate HIV-1 transcription. These results define a cellular kinase complex whose activity is modulated by Tat to result in activation of HIV-1 trancription.
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Quinasas Ciclina-Dependientes , Productos del Gen tat/metabolismo , VIH-1/genética , Proteínas Serina-Treonina Quinasas/metabolismo , ARN Polimerasa II/metabolismo , Factores de Transcripción TFII , Factores de Transcripción/metabolismo , Transcripción Genética , Diclororribofuranosil Benzoimidazol/farmacología , Regulación Viral de la Expresión Génica , Proteínas de Neoplasias/metabolismo , Inhibidores de la Síntesis del Ácido Nucleico/farmacología , Fosforilación , Factor B de Elongación Transcripcional Positiva , Unión Proteica , Proteínas Serina-Treonina Quinasas/aislamiento & purificación , Análisis de Secuencia , Factor de Transcripción TFIIH , Factores de Transcripción/aislamiento & purificación , Productos del Gen tat del Virus de la Inmunodeficiencia Humana , Quinasa Activadora de Quinasas Ciclina-DependientesRESUMEN
The ability of a human cytomegalovirus-encoded homolog of MHC class I molecules to serve as a peptide receptor was investigated. Sequencing of peptide material eluted from the purified viral protein revealed a mixture of endogenous peptides with characteristics similar to those eluted from conventional class I molecules, that is, anchor residues, and a predominance of short peptides derived from cytoplasmic proteins. The possible function(s) of this viral MHC homolog are discussed in light of the finding that it binds endogenous peptides.
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Citomegalovirus/química , Antígenos de Histocompatibilidad Clase I/química , Antígenos de Histocompatibilidad Clase I/fisiología , Proteínas Virales/química , Proteínas Virales/fisiología , Secuencia de Aminoácidos , Animales , Células CHO , Células Cultivadas , Cricetinae , Humanos , Modelos Moleculares , Datos de Secuencia Molecular , Unión Proteica/fisiología , TransfecciónRESUMEN
Five patients underwent pulmonary embolectomy without cardiopulmonary bypass. All had recent embolism completely obstructing the right branch of the artery but usually leaving the left branch open or almost totally free. In all patients fibrinolytic drugs were formally contra-indicated. Thoracotomy was performed through the right lateral approach. The embolus was completely removed; there were no post-operative complications and the results on follow-up are excellent.
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Arteria Pulmonar/cirugía , Embolia Pulmonar/cirugía , Adulto , Humanos , Persona de Mediana Edad , Embolia Pulmonar/diagnóstico por imagen , RadiografíaRESUMEN
The authors report two cases of traumatic rupture of the corpus cavernosum and have insisted on the early repair of the tunica albuginea best guarantee of adequate cure without erection impairement.
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Pene/lesiones , Adulto , Humanos , Masculino , Métodos , Pene/fisiología , Pene/cirugía , Rotura , Factores de TiempoRESUMEN
The authors report a case of bilharzia localised to one of the ureters in a case of double renal pelvis and ureter. The parasitic infection was identified only at the time of exploratory surgery (laboratory investigations and preoperative endoscopy negative). The "providential" double ureter made it possible to combine excision of the pathological ureter with a termino-lateral anastomosis between the lower renal pelvis and the ureter of the upper renal pelvis. The result remains very satisfactory with a follow-up of 13 months.