Cleavage specificity analysis of six type II transmembrane serine proteases (TTSPs) using PICS with proteome-derived peptide libraries.

Background: Type II transmembrane serine proteases (TTSPs) are a family of cell membrane tethered serine proteases with unclear roles as their cleavage site specificities and substrate degradomes have not been fully elucidated. Indeed just 52 cleavage sites are annotated in MEROPS, the database of proteases, their substrates and inhibitors.

Methodology/principal Finding: To profile the active site specificities of the TTSPs, we applied Proteomic Identification of protease Cleavage Sites (PICS).

Identification and relative quantification of native and proteolytically generated protein C-termini from complex proteomes: C-terminome analysis.

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.

Proteomic analyses reveal an acidic prime side specificity for the astacin metalloprotease family reflected by physiological substrates.

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.

Characterization of the prime and non-prime active site specificities of proteases by proteome-derived peptide libraries and tandem mass spectrometry.

To link cleaved substrates in complex systems with a specific protease, the protease active site specificity is required. Proteomic identification of cleavage sites (PICS) simultaneously determines both the prime- and non-prime-side specificities of individual proteases through identification of hundreds of individual cleavage sequences from biologically relevant, proteome-derived peptide libraries. PICS also identifies subsite cooperativity.

[Prevalence of Candida parapsilosis, C. orthopsilosis and C. metapsilosis in candidemia over a 5-year period at Nantes hospital and in vitro susceptibility to three echinocandins by E-test®].

Aim Of The Study: To determine the prevalence of C. parapsilosis sensu stricto, C. orthopsilosis and C.

Proteome-wide analysis of protein carboxy termini: C terminomics.

As proteome-wide C-terminal sequence analysis has been largely intractable, we developed a polymer-based enrichment approach to profile protein C-terminal peptides by mass spectrometry and identified hundreds of C-terminal peptides in the Escherichia coli proteome. We isotopically labeled GluC protease-digested and undigested samples and identified GluC substrates and their cleavage sites by quantification of neo-C-terminal peptides. Our method thus enables global annotation of protein C-terminal posttranslational modifications, including proteolytic truncations.

Characterization of the CopR regulon of Lactococcus lactis IL1403.

To identify components of the copper homeostatic mechanism of Lactococcus lactis, we employed two-dimensional gel electrophoresis to detect changes in the proteome in response to copper. Three proteins upregulated by copper were identified: glyoxylase I (YaiA), a nitroreductase (YtjD), and lactate oxidase (LctO). The promoter regions of these genes feature cop boxes of consensus TACAnnTGTA, which are the binding site of CopY-type copper-responsive repressors.

Copper induction of lactate oxidase of Lactococcus lactis: a novel metal stress response.

Lactococcus lactis IL1403, a lactic acid bacterium widely used for food fermentation, is often exposed to stress conditions. One such condition is exposure to copper, such as in cheese making in copper vats. Copper is an essential micronutrient in prokaryotes and eukaryotes but can be toxic if in excess.

Improved protocol for chromatofocusing on the ProteomeLab PF2D.

Beckman-Coulter has recently introduced the ProteomeLab PF2D for 2-D liquid separation of protein samples. The system features separation in the first dimension by chromatofocusing, followed by RP chromatography in the second dimension, allowing the analysis of complex proteomics samples. When used by the standard protocol, reproducibility and column life times are limited, making the use of the instrument very costly.

Characterization of the DNA-binding site in the ferric uptake regulator protein from Escherichia coli by UV crosslinking and mass spectrometry.

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.

New CT index to quantify arterial obstruction in pulmonary embolism: comparison with angiographic index and echocardiography.

Objective: This study was designed to define and evaluate a specific index to quantify arterial obstruction with helical CT in acute pulmonary embolism.

Materials And Methods: Fifty-four patients (mean age, 56 years) with proven pulmonary emboli among 158 consecutive patients, who had undergone both CT and pulmonary angiography for clinically suspected pulmonary embolism, were eligible for the study. The CT obstruction index was defined as (n.

Pulmonary embolism detection: prospective evaluation of dual-section helical CT versus selective pulmonary arteriography in 157 patients.

Purpose: To evaluate the accuracy of dual-section helical computed tomography (CT) in acute pulmonary embolism (PE) diagnosis.

Materials And Methods: Of 204 consecutive patients with clinically suspected acute PE (mean age, 58 years +/- 14 [SD]), 158 were enrolled. All patients underwent dual-section helical CT (2.

[Late solitary metastasis of a renal cancer to the contralateral adrenal gland].

We report a case of metachronous metastasis from renal cell carcinoma to the contralateral adrenal gland detected one year after radical nephrectomy. The initial tumor was incidentally discovered in the setting of acute aortic dissection. A large left adrenal tumor was detected on CT follow-up at two years.

[Bronchial and pulmonary vaso-occlusions].

The technique of vaso-occlusion (or embolisation) consists in occluding one or several vessels which are causing haemoptysis. Either of the pulmonary circulations (systemic bronchial or pulmonary) may be the cause and sometimes both are. In systemic pulmonary hypervascularization the pathological issues are affected by high pressure vascularisation at the expense of functional vascularisation and are the source of frequent episodes of bleeding.

Abdominal aortic aneurysm: pretherapy assessment with dual-slice helical CT angiography.

Objective: The purpose of this study was to evaluate dual-slice helical CT in the pretherapy assessment of abdominal aortic aneurysms.

Subjects And Methods: 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).

[Arterial lipiodol chemoembolization of a left liver metastasis through the right internal thoracic artery].

The purpose of this article is to illustrate the efficacy of the chemoembolization in patients with hypervascular metastases and to describe the post-embolization change in vascularization pattern. Unusual collaterals may develop following embolization. A 59-year-old woman, followed for unresectable small bowel carcinoid tumor since 1991, underwent successful chemoembolization of several liver metastases.

Percutaneous gastrostomy for enteral nutrition: long-term follow-up of 176 procedures.

Objective: To evaluate the efficacy and safety of percutaneous gastrostomy (PG), using small catheters without gastropexy, to deliver enteral nutrition.

Methods: We reviewed the records of 176 consecutive patients in whom PG was attempted for enteral nutrition. Of these cases, 172 catheters were inserted by the Seldinger technique under fluoroscopic guidance alone, 2 were inserted under computed tomographic guidance, and 2 procedures failed.

Subacute and chronic benign superior vena cava obstructions: endovascular treatment with self-expanding metallic stents.

Objective: 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).

Subjects And Methods: 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.

[Preoperative angiographic landmarks of the gluteal muscle regions using Patent Blue V].

Use of a gluteus musculocutaneous flap is the most reliable technique for surgical repair of sacral ulcers. Surgery could be ideally performed when the flap is designed using only the upper or the lower half of the glutens maximus muscle depending on the superior gluteal artery or the inferior gluteal artery, respectively. The authors have developed a technique for preoperative demonstration of the vascular supply to both muscle and overlying skin from a single gluteal artery using superselective catheterization and arterial administration of a dye (Patent Blue V).

Helical CT phlebography of the superior vena cava: diagnosis and evaluation of venous obstruction.

Objective: 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.

Subjects And Methods: 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.

[Positional occlusion of the descending scapular artery in transverse thoraco-brachial syndrome with neurologic symptoms].

We report a thoracic outlet syndrome revealed by neurological complications. Angiography of the subclavian artery depicted an isolated positional occlusion of the descending scapular artery. This side branch of the subclavian artery is anatomically located close to and supplies the brachial plexus.

The comet-tail artifact. An ultrasound sign of alveolar-interstitial syndrome.

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.