Astacins are secreted and membrane-bound metalloproteases with clear associations to many important pathological and physiological processes. Yet with only a few substrates described their biological ...roles are enigmatic. Moreover, the lack of knowledge of astacin cleavage site specificities hampers assay and drug development. Using PICS (proteomic identification of protease cleavage site specificity) and TAILS (terminal amine isotopic labeling of substrates) degradomics approaches >3000 cleavage sites were proteomically identified for five different astacins. Such broad coverage enables family-wide determination of specificities N- and C-terminal to the scissile peptide bond. Remarkably, meprin α, meprin β, and LAST_MAM proteases exhibit a strong preference for aspartate in the peptide (P)1′ position because of a conserved positively charged residue in the active cleft subsite (S)1′. This unparalleled specificity has not been found for other families of extracellular proteases. Interestingly, cleavage specificity is also strongly influenced by proline in P2′ or P3′ leading to a rare example of subsite cooperativity. This specificity characterizes the astacins as unique contributors to extracellular proteolysis that is corroborated by known cleavage sites in procollagen I+III, VEGF (vascular endothelial growth factor)-A, IL (interleukin)-1β, and pro-kallikrein 7. Indeed, cleavage sites in VEGF-A and pro-kallikrein 7 identified by terminal amine isotopic labeling of substrates matched those reported by Edman degradation. Moreover, the novel substrate FGF-19 was validated biochemically and shown to exhibit altered biological activity after meprin processing.
In the design process of any device, the computation of the forces or the torque is often a decisive step. Finite element solutions give accurate solutions in terms of magnetic field or flux density. ...But for local force computation associated to a magnetic field, many formulations are available. They use different physical interpretations of the same phenomenon. Here, a new experiment is presented. A test body is submitted to an external field and its deformation is observed. With an unusual material, the effects of external strains induced by magnetic or electric fields are increased. Numerical simulations allow the calculation of local forces associated to all formulations. Consequently it is possible to use them to deduce the body deformations. It appears that the body deformation associated to the local force provided by the energy principle corresponds to the experiment
Mutual inductance is a phenomenon caused by the circulation of the magnetic flux in the core of an electrical machine. It is the result of the effect of the current flowing in one phase on the other ...phases. In conventional three-phase machines, such an effect has no influence on the electrical behaviour of the device. Although these machines are powered by power inverters, no problem should occur. The result is not the same for multi-star machines. If these machines are using a conventional winding structure, namely distributed windings, and are powered by voltage source converters, current ripples appear in the power supply lines. These current ripples are related to magnetic couplings between the stars. Designers should check these current ripples in order to stay within the limits imposed by the specifications. These electric current disturbances also provide torque ripples. With concentrated windings, a new degree of freedom appears; the configuration-number of slots/number of poles-can have a positive impact. The circulation of the magnetic flux is the initial phenomenon that produces the mutual inductance. The main goal of this discussion is to describe a design method that is able to produce not only a machine with low mutual inductance between phases, but also a multi-star machine where the stars and the phases are magnetically decoupled or less coupled. This discussion only takes into account the machines that use permanent magnets mounted on the rotor surface. This article is part of a study aimed at designing a high efficiency generator using fractional-slot concentrated-windings (FSCW).
Proteome-wide analysis of protein C-termini has long been inaccessible, but is now enabled by a newly developed negative selection strategy we term C-terminomics. In this procedure, amine- and ...carboxyl groups of full-length proteins are chemically protected. After trypsin digestion, N-terminal and internal tryptic peptides - but not C-terminal peptides - posses newly formed, unprotected C-termini that are removed by coupling to the high-molecular-weight polymer poly-allylamine. Ultrafiltration separates the uncoupled, blocked C-terminal peptides that are subsequently analyzed by liquid chromatography-tandem mass spectrometry. On a proteome-wide scale, this strategy profiles native protein C-termini together with neo C-termini generated by endoproteolytic cleavage or processive C-terminal truncations ("ragging"). In bacterial proteomes, hundreds of protein C-termini were identified. Stable isotope labeling enables -quantitative comparison of protein C-termini and C-terminal processing in different samples. Using formaldehyde-based chemical labeling, this quantitative approach termed "carboxy-terminal amine-based isotope labeling of substrates (C-TAILS)" identified >100 cleavage sites of exogenously applied GluC protease in an Escherichia coli proteome. C-TAILS complements recently developed N-terminomic techniques for endoprotease substrate discovery and is essential for the characterization of carboxyprotease processing.
Can ultrasound be of any help in the diagnosis of alveolar-interstitial syndrome? In a prospective study, we examined 250 consecutive patients in a medical intensive care unit: 121 patients with ...radiologic alveolar-interstitial syndrome (disseminated to the whole lung, n = 92; localized, n = 29) and 129 patients without radiologic evidence of alveolar-interstitial syndrome. The antero-lateral chest wall was examined using ultrasound. The ultrasonic feature of multiple comet-tail artifacts fanning out from the lung surface was investigated. This pattern was present all over the lung surface in 86 of 92 patients with diffuse alveolar-interstitial syndrome (sensitivity of 93.4%). It was absent or confined to the last lateral intercostal space in 120 of 129 patients with normal chest X-ray (specificity of 93.0%). Tomodensitometric correlations showed that the thickened sub-pleural interlobular septa, as well as ground-glass areas, two lesions present in acute pulmonary edema, were associated with the presence of the comet-tail artifact. In conclusion, presence of the comet-tail artifact allowed diagnosis of alveolar-interstitial syndrome.
The purpose of this study was to evaluate dual-slice helical CT in the pretherapy assessment of abdominal aortic aneurysms.
Dual-slice helical CT angiography was performed in 47 consecutive patients ...(mean age, 59 years) with abdominal aortic aneurysm to determine whether we could then evaluate the extent of aneurysm and see associated renal, celiac, mesenteric, and iliofemoral artery disease. Results were compared with those of digital subtraction angiography (n = 47) and surgery (n = 37).
The proximal and distal extents of abdominal aortic aneurysm correlated well with surgical findings. Dual-slice helical CT showed all main (n = 102) and accessory (n = 13) renal arteries with a sensitivity of 91% and a specificity of 100% for revealing associated renal artery stenosis exceeding 50%. Sensitivity and specificity of dual-slice helical CT for revealing stenosis exceeding 75% in celiac and superior mesenteric arteries were both 100%. Three of four iliofemoral artery stenoses and two occlusions of the common iliac artery were revealed by dual-slice helical CT.
Helical CT angiography with dual-slice scanning is a useful and minimally invasive technique that can provide with high accuracy all the necessary information for treatment of abdominal aortic aneurysm.
Ferric uptake regulator protein (Fur) is activated by its cofactor iron to a state that binds to a specific DNA sequence called ‘Fur box’. Using mass spectrometry-based methods, we showed that Tyr 55 ...of
Escherichia coli Fur, as well as the two thymines in positions 18 and 19 of the consensus Fur Box, are involved with binding. A conformational model of the Fur–DNA complex is proposed, in which DNA is in contact with each H4 A52–A64 Fur helix. We propose that this interaction is a common feature for the Fur-like proteins, such as Zur and PerR, and their respective DNA boxes.
Our purpose is to report our clinical experience with patients who underwent endovascular treatment with Wallstents for subacute or chronic benign obstruction of the superior vena cava (SVC).
Twelve ...patients who were an average of 54 +/- 12 years old were referred for treatment of severe SVC syndrome related to implanted central venous catheters (n = 8), postradiation fibrosis (n = 2), a permanent pacemaker (n = 1), or a benign tumor (n = 1). Symptoms were present for an average of 16 weeks (range, 4-48 weeks) before treatment. Diagnosis of SVC obstruction was confirmed with helical CT and pretherapeutic phlebography. Four patients had Stanford's type II stenosis; two, type III; and six, type IV. The mean clinical and radiologic follow-up intervals were 11 months (range, 1-36 months) and 7 months (range, 1 week to 32 months), respectively.
Recanalization was successful in all patients. Fifteen stents were implanted in the 12 patients. Stents were placed after percutaneous balloon angioplasty in nine patients, and primary stent placement was attempted in three patients. We immediately achieved a satisfactory SVC diameter in all patients, whose symptoms were relieved completely within 1 week of stent placement. No technical or clinical complications occurred. SVC syndrome recurred in one patient 2 months after stent placement and was treated by placing a second stent.
Endovascular treatment with stent placement should be considered relevant and safe for refractory benign SVC syndrome. However, a larger series and a longer follow-up period are needed to define the role of stent placement for this syndrome.
The purpose of this study was to determine the feasibility of helical CT phlebography of the superior vena cava (SVC) and to evaluate the role of this imaging technique in the diagnosis and treatment ...of SVC obstruction.
Twenty-three helical CT phlebograms were obtained of patients with clinical findings that were suggestive of SVC obstruction (n = 19) and of patients undergoing posttherapeutic evaluation for SVC obstruction (n = 4). CT examinations consisted of helical acquisitions obtained in the craniocaudal direction with simultaneous bilateral antecubital vein injection of 2 x 90 ml of 12% iodinated contrast material at 2 ml/sec. Combined analysis of axial, multiplanar, and maximum-intensity-projection reformatted images was used for all patients. Image quality, venous stenosis or obstruction, intraluminal thrombus, and collateral pathways were evaluated. Comparison with digital phlebographic data was available for 16 patients; this comparison was performed in a nonblinded manner.
CT phlebograms were considered technically optimal in 91% of the patients. In all these patients, helical CT phlebograms showed the venous obstruction: the site, extent, cause, and collateral pathways. CT phlebography appeared to be well correlated with digital phlebography in 16 patients regarding the degree of obstruction, the presence of collateral pathways, and the presence of thrombus.
Helical CT phlebography may be a useful technique for imaging the SVC and its tributaries. This imaging technique is simple to perform and can provide all the information necessary to diagnose and treat SVC obstruction.
PDF
