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21.
Comparison of two ELISA methods for measuring levels of the phosphorylated neurofilament heavy chain
BACKGROUND: Recent studies suggest that the quantification of neurofilament subunits in cerebrospinal fluid (CSF), blood and amniotic fluid may reflect neuroaxonal damage and be of high clinical value. The present study aims to cross-validate two different independently developed ELISA techniques for the quantification of the phosphorylated axonal forms of the neurofilament heavy chain (pNfH). METHODS: The London ELISA method is based on barbitone buffer and the commercially available SMI35 capture antibody. The Gainesville method uses Tris-buffered saline (TBS) and an affinity purified chicken polyclonal capture antibody (C-pNfH). Coded CSF from 50 patients with neurological diseases were analyzed in duplicate by both laboratories, each using both ELISA methods, but with each lab using their own detection antibody, tertiary antibody and chromogen. Methods were compared using Bland-Altman plots. Correlation and regression analyses were used to allow for transformation of values between both methods. RESULTS: The Bland-Altman plots demonstrated that 96% of all samples fell into the narrow 95% limits of agreement (0.04 units of OD). There was a high correlation (Spearman R=0.92, p<0.0001 and Pearson R=0.98, p<0.0001) between the Gainesville (Y) and the London (X) method with Y=0.132+1.104(*)X. The previously determined upper reference limit of 0.73 mg/l (London method) corresponds to 0.94 mg/l for the Gainesville method. CSF pNfH levels above the reference limit were observed in patients with encephalitis, encephalomyelitis, hydrocephalus, subarachnoid haemorrhage, spino-muscular atrophy, stroke and cancer with both methods agreeing in all cases. CONCLUSION: The two assays are in excellent agreement, suggesting that pNfH, which has a number of unusual protein chemical features, may be the biomarker of choice for the routine and robust detection of axonal injury and degeneration in both research and clinical contexts. 相似文献
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Prof. Dr. T. Schmitz-Rixen F. Adili R. Bauersachs I. Eichler J. Hoffmann J. Klocker J. Kobba T. Petzold H. Wenk R.T. Grundmann 《Gef?sschirurgie》2014,19(7):667-674
Background
The direct thrombin inhibitor argatroban is available for parenteral antithrombotic treatment of patients with heparin-induced thrombocytopenia type II (HIT II).Method
Experiences with argatroban were exchanged in a workshop based on case reports and a survey of experts. The aim was to define the indications and modalities for administration of argatroban in vascular surgery.Results
The recommended body weight-related therapeutic intravenous (i.v.) dosage of argatroban is 0.5–2 µg/kg/min. The therapy can be monitored by the activated partial thromboplastin time (aPTT) with a target value of a 1.5-fold to 3-fold prolongation of the aPTT. A dose reduction is required in patients with compromised liver function. If anticoagulation is only to be carried out intraoperatively, the administration of a bolus immediately before clamping of the vessels (e.g. carotid artery or aorta) is possible (bolus of 5–15 mg depending on body weight, preferred 10 mg). The treatment can then be continued with an infusion of 0.5–2 µg/kg/min, whereby the daily dosage has to be considered (for a 70 kg patient approximately 50–200 mg).Conclusion
Uncertainties exist in the administration of argatroban. The dosages recommended here have a substantial range depending on the cardiac function of the patient, liver function and whether the patient needs intensive care unit (ICU) treatment. The dosages should then be rechecked. The Research Committee of the German Society for Vascular Surgery therefore proposes that in the future HIT II patients should be recorded in a register. Due to the rare incidence of the disease all surgical ICUs are invited to participate. 相似文献23.
Phenotype of mice with inducible ablation of GluA1 AMPA receptors during late adolescence: Relevance for mental disorders
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Tillmann Weber Juan M. Lima‐Ojeda Miriam Schneider Alessia Luoni Marco A. Riva Karen Gertz Julian Hellmann‐Regen Golo Kronenberg Andreas Meyer‐Lindenberg Rolf Sprengel Peter Gass 《Hippocampus》2014,24(4):424-435
Adolescence is characterized by important molecular and anatomical changes with relevance for the maturation of brain circuitry and cognitive function. This time period is of critical importance in the emergence of several neuropsychiatric disorders accompanied by cognitive impairment, such as affective disorders and schizophrenia. The molecular mechanisms underlying these changes at neuronal level during this specific developmental stage remains however poorly understood. GluA1‐containing AMPA receptors, which are located predominantly on hippocampal neurons, are the primary molecular determinants of synaptic plasticity. We investigated here the consequences of the inducible deletion of GluA1 AMPA receptors in glutamatergic neurons during late adolescence. We generated mutant mice with a tamoxifen‐inducible deletion of GluA1 under the control of the CamKII promoter for temporally and spatially restricted gene manipulation. GluA1 ablation during late adolescence induced cognitive impairments, but also marked hyperlocomotion and sensorimotor gating deficits. Unlike the global genetic deletion of GluA1, inducible GluA1 ablation during late adolescence resulted in normal sociability. Deletion of GluA1 induced redistribution of GluA2 subunits, suggesting AMPA receptor trafficking deficits. Mutant animals showed increased hippocampal NMDA receptor expression and no change in striatal dopamine concentration. Our data provide new insight into the role of deficient AMPA receptors specifically during late adolescence in inducing several cognitive and behavioral alterations with possible relevance for neuropsychiatric disorders. © 2013 Wiley Periodicals, Inc. 相似文献
24.
Lefetamine‐derived designer drugs N‐ethyl‐1,2‐diphenylethylamine (NEDPA) and N‐iso‐propyl‐1,2‐diphenylethylamine (NPDPA): Metabolism and detectability in rat urine using GC‐MS,LC‐MSn and LC‐HR‐MS/MS
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Carina S. D. Wink Golo M. J. Meyer Dirk K. Wissenbach Andrea Jacobsen‐Bauer Markus R. Meyer Hans H. Maurer 《Drug testing and analysis》2014,6(10):1038-1048
N‐Ethyl‐1,2‐diphenylethylamine (NEDPA) and N‐iso‐propyl‐1,2‐diphenylethylamine (NPDPA) are two designer drugs, which were confiscated in Germany in 2008. Lefetamine (N,N‐dimethyl‐1,2‐diphenylethylamine, also named L‐SPA), the pharmaceutical lead of these designer drugs, is a controlled substance in many countries. The aim of the present work was to study the phase I and phase II metabolism of these drugs in rats and to check for their detectability in urine using the authors’ standard urine screening approaches (SUSA). For the elucidation of the metabolism, rat urine samples were worked up with and without enzymatic cleavage, separated and analyzed by gas chromatography‐mass spectrometry (GC‐MS) and liquid chromatography‐high resolution‐tandem mass spectrometry (LC‐HR‐MS/MS). According to the identified metabolites, the following metabolic pathways for NEDPA and NPDPA could be proposed: N‐dealkylation, mono‐ and bis‐hydroxylation of the benzyl ring followed by methylation of one of the two hydroxy groups, combinations of these steps, hydroxylation of the phenyl ring after N‐dealkylation, glucuronidation and sulfation of all hydroxylated metabolites. Application of a 0.3 mg/kg BW dose of NEDPA or NPDPA, corresponding to a common lefetamine single dose, could be monitored in rat urine using the authors’ GC‐MS and LC‐MSn SUSA. However, only the metabolites could be detected, namely N‐deethyl‐NEDPA, N‐deethyl‐hydroxy‐NEDPA, hydroxy‐NEDPA, and hydroxy‐methoxy‐NEDPA or N‐de‐iso‐propyl‐NPDPA, N‐de‐iso‐propyl‐hydroxy‐NPDPA, and hydroxy‐NPDPA. Assuming similar kinetics, an intake of these drugs should also be detectable in human urine. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
25.
Bergman E Kieler H Petzold M Sonesson C Axelsson O 《Acta obstetricia et gynecologica Scandinavica》2007,86(6):671-677
BACKGROUND: Antenatal identification of infants small for gestational age (SGA) improves their perinatal outcome. Repeated measurement of symphysis-fundus (SF) heights performed by midwives is the most widespread screening method for detection of SGA. However, the inefficiency of this method necessitates improved practices. Earlier start and more frequent SF measurements, which could be accomplished by self-administered measurements, might improve the ability to detect deviant growth. The present study was set up to evaluate whether pregnant women can reliably perform SF measurements by themselves. METHOD: Forty healthy women with singleton and ultrasound-dated pregnancies from 2 antenatal clinics in Uppsala, Sweden, were asked to perform 4 consecutive SF measurements once every week, from 20 to 25 weeks of gestation until delivery. The self-administered SF measurements were recorded and systematically compared with midwives' SF measurements. RESULTS: Thirty-three pregnant women performed self-administered SF measurements over a 14-week period (range: 1-21). The SF curves constructed from self-administered SF measurements had the same shape as previously constructed population-based reference curves. The variance for self-administered SF measurements was higher than that of the midwives. CONCLUSIONS: Pregnant women are capable of measuring SF heights by themselves, but with higher individual variance than midwives. Repeated measurements at each occasion can compensate for the higher variance. The main advantage of self-administered SF measurements is the opportunity to follow fetal growth earlier and more frequently. 相似文献
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Christoph Wagener Petra Petzold Wolfgang Khler Vladimir Totovi 《International journal of cancer. Journal international du cancer》1984,33(4):469-475
Five monoclonal antibodies that recognize different epitopes on carcinoembryonic antigen (CEA) were reacted with tissue sections of various carcinoma specimens. The vital, non-necrotic tissues of those carcinomas of the oesophagus, pancreas, colon and rectum, medullary thyroid, ovary and cervix in which CEA related epitopes were detectable bound all five antibodies to a similar degree. In 3/5 lung carcinomas, 2/10 mammary carcinomas and 1/5 gastric carcinomas, a significantly different binding of the monoclonal antibodies by vital tumour tissue was present as determined by three independent investigators. In necrotic tissue areas, heterogeneity of antibody binding was more common. In a previous investigation, it was shown that normal granulocytes and liver tissue differentially bind the monoclonal anti-CEA antibodies, indicating the presence of crossreacting antigens. The equivalent binding of the five monoclonal anti-CEA antibodies to most of the carcinomas tested suggests that this binding is due to the presence of complete CEA molecules and not only of cross-reacting antigens. 相似文献
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