64 research outputs found
Silver(i) Perfluoroalcoholates: Synthesis, Structure, and their Use as Transfer Reagents
Herein we report a general access to silver(i) perfluoroalcoholates, their structure in the solid state and in solution, and their use as transfer reagents. The silver(i) perfluoroalcoholates are prepared by the reaction of AgF with the corresponding perfluorinated carbonyl compounds in acetonitrile and are stable for a prolonged time at –18 °C. X-Ray analysis of single crystals of perfluoroalcoholate species showed that two Ag(i) centers are bridged by the alcoholate ligands. In acetonitrile solution, Ag[OCF3] forms different structures as indicated by IR spectroscopy. Furthermore, the silver(i) perfluoroalcoholates can be used as easy-to-handle transfer reagents for the synthesis of Cu[OCF3], Cu[OC2F5], [PPh4][Au(CF3)3(OCF3)], and fluorinated alkyl ethers
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Physiologic artifacts in resting state oscillations in young children: methodological considerations for noisy data
We quantified the potential effects of physiologic artifact on the estimation of EEG band power in a cohort of typically developing children in order to guide artifact rejection methods in quantitative EEG data analysis in developmental populations. High density EEG was recorded for 2 min while children, ages 2–6, watched a video of bubbles. Segments of data were categorized as blinks, saccades, EMG or artifact-free, and both absolute and relative power in the theta (4–7 Hz), alpha (8–12 Hz), beta (13–30 Hz) and gamma (35–45 Hz) bands were calculated in 9 regions for each category. Using a linear mixed model approach with artifact type, region and their interaction as predictors, we compared mean band power between clean data and each type of artifact. We found significant differences in mean relative and absolute power between artifacts and artifact-free segments in all frequency bands. The magnitude and direction of the differences varied based on power type, region, and frequency band. The most significant differences in mean band power were found in the gamma band for EMG artifact and the theta band for ocular artifacts. Artifact detection strategies need to be sensitive to the oscillations of interest for a given analysis, with the most conservative approach being the removal of all EMG and ocular artifact from EEG data. Quantitative EEG holds considerable promise as a clinical biomarker of both typical and atypical development. However, there needs to be transparency in the choice of power type, regions of interest, and frequency band, as each of these variables are differentially vulnerable to noise, and therefore, their interpretation depends on the methods used to identify and remove artifacts
Generating Antibodies Against Secreted Proteins Using Vascular Smooth Muscle Cells Transduced with Replication-Defective Retrovirus
The traditional method of antibody (Ab) generation requires repeated injections of antigen (Ag). We have developed an alternative method that allows an investigator to generate a polyclonal antiserum with only a cDNA in hand. We cloned a cDNA encoding the coding frame for baboon tissue inhibitor of matrix metalloproteinase-1 (TIMP-1). Fischer rat arterial smooth muscle cells (SMC) transduced with the baboon TIMP-1 using a replication-defective retrovirus were propagated in culture. TIMP-1 overexpressing rat SMC were seeded into deendothelialized rat carotid arteries. Three weeks after cell seeding in the rat, the presence of Ab to the baboon TIMP-1 was detected by dot blot and enzyme-linked immunosorbent assay in 5 of 6 of the animals. The major portion of the Ab generated against baboon TIMP-1 during a 12-month monitoring period afer the cell seeding was identified as belonging to the IgGl subtype. More interestingly, the titer of the Ab kept rising throughout an 8-month monitoring period. Among the salient features of this Ab are its capacily to block TIMP-1 activity and its utility for detecting TIMP-1 by immunohistochemistry. These results demonstrate that Ab against a secreted protein can be obtained in response to continuous expression of the cDNA by vascular SMC. Purified Ag is not required
Prevention of Aneurysm Development and Rupture by Local Overexpression of Plasminogen Activator Inhibitor-1
Background
—Arterial aneurysms exhibit a loss of elastin and an increase in the plasminogen activators urokinase plasminogen activator (u-PA) and tissue plasminogen activator (t-PA). Because u-PA, t-PA, and plasmin have a limited proteolytic activity against elastin, the role of plasminogen activators in the aneurysmal disease is unclear. To investigate this question, we overexpressed plasminogen activator inhibitor-1 (PAI-1), an inhibitor of t-PA and u-PA, in a rat model of aortic aneurysm.
Methods and Results
—Guinea pig–to-rat aortic xenografts were seeded with syngeneic Fischer 344 rat smooth muscle cells retrovirally transduced with the rat PAI-1 gene (LPSN group) or the vector alone (LXSN group). Some grafts were not seeded with cells (NO group). Western blots showed increased PAI-1 in grafts from the LPSN group compared with LXSN and NO groups. All grafts in the NO group (n=8) and 40% in the LXSN group ruptured between days 4 and 14. At 4 weeks in the LXSN group, the remaining unruptured grafts (n=6) were aneurysmal (diameter increase ≥100%), whereas in the LPSN group (n=6) none of the grafts had ruptured or were aneurysmal. Elastin was preserved in the LPSN group. t-PA, the major PA expressed in the model, was decreased in the LPSN group compared with the other groups, as determined by zymography. Quantitative zymography showed decreased levels of two matrix metalloproteinases (MMPs), a 28-kD caseinase, and activated MMP-9 in the LPSN group.
Conclusions
—The blockade of plasminogen activators prevents formation of aneurysms and arterial rupture by inhibiting MMP activation.
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