Mutant and chimeric recobinant plasminogen activatorsproduction in eukaryotic cellsand preliminary characterization

Mutant urokinase-type plasminogen activator (u-PA) genes and hybrid genes between tissue-type plasminogen activator (t-PA) and u-PA have been designed to direct the synthesis of new plasminogen activators and to investigate the structure-function relationship in these molecules. The following classes of constructs were made starting from cDNA encoding human t-PA or u-PA: 1) u-PA mutants in which the Arg156 and Lys158 were substituted with threonine, thus preventing cleavage by thrombin and plasmin; 2) hybrid molecules in which the NH2-terminal regions of t-PA (amino acid residues 1-67, 1-262, or 1-313) were fused with the COOH-terminal region of u-PA (amino acids 136-411, 139-411, or 195-411, respectively); and 3) a hybrid molecule in which the second kringle of t-PA (amino acids 173-262) was inserted between amino acids 130 and 139 of u-PA. In all cases but one, the recombinant proteins, produced by transfected eukaryotic cells, were efficiently secreted in the culture medium. The translation products have been tested for their ability to activate plasminogen after in situ binding to an insolubilized monoclonal antibody directed against urokinase. All recombinant enzymes were shown to be active, except those in which Lys158 of u-PA was substituted with threonine. Recombination of structural regions derived from t-PA, such as the finger, the kringle 2, or most of the A-chain sequences, with the protease part or the complete u-PA molecule did not impair the catalytic activity of the hybrid polypeptides. This observation supports the hypothesis that structural domains in t-PA and u-PA fold independently from one to another

Stratification of single-vessel coronary stenosis by ischemic threshold at the onset of wall motion abnormality during continuous monitoring of left ventricular function by semisupine exercise echocardiography.

peer reviewedWe studied the relation between the ischemic threshold at the onset of wall motion abnormality on exercise echocardiography (EE) and the severity of coronary stenosis in patients with 1-vessel coronary artery disease (CAD). We screened 216 consecutive patients who underwent coronary angiography and EE for suspected CAD. Ninety-five (74 men; age, 56 +/- 12 years) satisfied the study criteria, that is, the presence of 1-vessel disease or no evidence of CAD on angiography and a normal baseline echocardiogram. Eighty-seven patients had 1-vessel CAD on angiography, and exercise-induced wall motion abnormality occurred in 73 (77%). Optimal cutoff values of percent diameter stenosis and minimal lumen diameter for predicting a positive EE were 61% (sensitivity and specificity of 76%) and 1.12 mm (sensitivity and specificity of 74%). Among patients with positive EE, heart rate-blood pressure product at ischemic threshold was correlated with quantitative coronary stenosis (r = -0.72, P <.001). The ischemic threshold from continuous monitoring of left ventricular function during semisupine EE is correlated with the severity of coronary stenosis among patients with 1-vessel disease and a normal resting echocardiogram

How I treat ... the athlete's foot and its non-mycotic cutaneous pathology

peer reviewedSkin and nails of the foot of sport practitioners of various disciplines are subjected to the effects of benign but invalidating pathologies. Microtraumatisms are frequently involved. Beside dermatomycoses and onychomycoses, a dozen of typical disorders are identified

MRSA clonal complex 22 strains harboring toxic shock syndrome toxin (TSST-1) are endemic in the primary hospital in Gaza, Palestine

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen in both community and healthcare-related settings worldwide. Current knowledge regarding the epidemiology of S. aureus and MRSA in Gaza is based on a single community-based carriage study. Here we describe a cross-sectional analysis of 215 clinical isolates collected from Al-Shifa Hospital in Gaza during 2008 and 2012. METHODS: All isolates were characterized by spa typing, SCCmec typing, and detection of genes encoding Panton-Valentine leukocidin (PVL) and toxic shock syndrome toxin (TSST-1). Representative genotypes were also subjected to multilocus sequence typing (MLST). Antibiotic susceptibility testing was performed using VITEK2 and MicroScan. RESULTS: MRSA represented 56.3% of all S. aureus strains, and increased in frequency from 2008 (54.8%) to 2012 (58.4%). Aside from beta-lactams, resistance was observed to tetracycline, erythromycin, clindamycin, gentamicin, and fluoroquinolones. Molecular typing identified 35 spa types representing 17 MLST clonal complexes (CC), with spa 998 (Ridom t223, CC22) and spa 70 (Ridom t044, CC80) being the most prevalent. SCCmec types I, III, IV, V and VI were identified among MRSA isolates, while type II was not detected. PVL genes (lukF/S-PV) were detected in 40.0% of all isolates, while the TSST-1 gene (tst) was detected in 27.4% of all isolates, with surprisingly high frequency within CC22 (70.4%). Both PVL and TSST-1 genes were found in several isolates from 2012. CONCLUSIONS: Molecular typing of clinical isolates from Gaza hospitals revealed unusually high prevalence of TSST-1 genes among CC22 MRSA, which is noteworthy given a recent community study describing widespread carriage of a CC22 MRSA clone known as the \u27Gaza strain\u27. While the latter did not address TSST-1, tst-positive spa 998 (Ridom t223) has been detected in several neighboring countries, and described as endemic in an Italian NICU, suggesting international spread of a \u27Middle Eastern variant\u27 of pandemic CC22 strain EMRSA-15

Outcomes of Patients with Asymptomatic Aortic Stenosis Followed Up in Heart Valve Clinics

Importance: The natural history and the management of patients with asymptomatic aortic stenosis (AS) have not been fully examined in the current era. Objective: To determine the clinical outcomes of patients with asymptomatic AS using data from the Heart Valve Clinic International Database. Design, Setting, and Participants: This registry was assembled by merging data from prospectively gathered institutional databases from 10 heart valve clinics in Europe, Canada, and the United States. Asymptomatic patients with an aortic valve area of 1.5 cm2 or less and preserved left ventricular ejection fraction (LVEF) greater than 50% at entry were considered for the present analysis. Data were collected from January 2001 to December 2014, and data were analyzed from January 2017 to July 2018. Main Outcomes and Measures: Natural history, need for aortic valve replacement (AVR), and survival of asymptomatic patients with moderate or severe AS at entry followed up in a heart valve clinic. Indications for AVR were based on current guideline recommendations. Results: Of the 1375 patients included in this analysis, 834 (60.7%) were male, and the mean (SD) age was 71 (13) years. A total of 861 patients (62.6%) had severe AS (aortic valve area less than 1.0 cm2). The mean (SD) overall survival during medical management (mean [SD] follow up, 27 [24] months) was 93% (1%), 86% (2%), and 75% (4%) at 2, 4, and 8 years, respectively. A total of 104 patients (7.6%) died under observation, including 57 patients (54.8%) from cardiovascular causes. The crude rate of sudden death was 0.65% over the duration of the study. A total of 542 patients (39.4%) underwent AVR, including 388 patients (71.6%) with severe AS at study entry and 154 (28.4%) with moderate AS at entry who progressed to severe AS. Those with severe AS at entry who underwent AVR did so at a mean (SD) of 14.4 (16.6) months and a median of 8.7 months. The mean (SD) 2-year and 4-year AVR-free survival rates for asymptomatic patients with severe AS at baseline were 54% (2%) and 32% (3%), respectively. In those undergoing AVR, the 30-day postprocedural mortality was 0.9%. In patients with severe AS at entry, peak aortic jet velocity (greater than 5 m/s) and LVEF (less than 60%) were associated with all-cause and cardiovascular mortality without AVR; these factors were also associated with postprocedural mortality in those patients with severe AS at baseline who underwent AVR (surgical AVR in 310 patients; transcatheter AVR in 78 patients). Conclusions and Relevance: In patients with asymptomatic AS followed up in heart valve centers, the risk of sudden death is low, and rates of overall survival are similar to those reported from previous series. Patients with severe AS at baseline and peak aortic jet velocity of 5.0 m/s or greater or LVEF less than 60% have increased risks of all-cause and cardiovascular mortality even after AVR. The potential benefit of early intervention should be considered in these high-risk patients

Bloodstream and endovascular infections due to Abiotrophia defectiva and Granulicatella species

BACKGROUND: Abiotrophia and Granulicatella species, previously referred to as nutritionally variant streptococci (NVS), are significant causative agents of endocarditis and bacteraemia. In this study, we reviewed the clinical manifestations of infections due to A. defectiva and Granulicatella species that occurred at our institution between 1998 and 2004. METHODS: The analysis included all strains of NVS that were isolated from blood cultures or vascular graft specimens. All strains were identified by 16S rRNA sequence analysis. Patients' medical charts were reviewed for each case of infection. RESULTS: Eleven strains of NVS were isolated during the 6-year period. Identification of the strains by 16S rRNA showed 2 genogroups: Abiotrophia defectiva (3) and Granulicatella adiacens (6) or "para-adiacens" (2). The three A. defectiva strains were isolated from immunocompetent patients with endovascular infections, whereas 7 of 8 Granulicatella spp. strains were isolated from immunosuppressed patients, mainly febrile neutropenic patients. We report the first case of "G. para-adiacens" bacteraemia in the setting of febrile neutropenia. CONCLUSION: We propose that Granulicatella spp. be considered as a possible agent of bacteraemia in neutropenic patients

Fluid dynamic lateral slicing of high tensile strength carbon nanotubes

Lateral slicing of micron length carbon nanotubes (CNTs) is effective on laser irradiation of the materials suspended within dynamic liquid thin films in a microfluidic vortex fluidic device (VFD). The method produces sliced CNTs with minimal defects in the absence of any chemical stabilizers, having broad length distributions centred at ca 190, 160 nm and 171 nm for single, double and multi walled CNTs respectively, as established using atomic force microscopy and supported by small angle neutron scattering solution data. Molecular dynamics simulations on a bent single walled carbon nanotube (SWCNT) with a radius of curvature of order 10 nm results in tearing across the tube upon heating, highlighting the role of shear forces which bend the tube forming strained bonds which are ruptured by the laser irradiation. CNT slicing occurs with the VFD operating in both the confined mode for a finite volume of liquid and continuous flow for scalability purposes

Wood biscuit measuring and sorting system

V diplomskem delu je predstavljena izdelava naprave za dimenzijsko kontrolo lesenih veznih lamel. Opisana je avtomatska mehanska linija naprave, ki zajema zalogovnik, dvigalo, rotacijski vibracijski dodajalnik, merilno mesto ter sistem za ločevanje dobrih in slabih lamel. Osrednji del diplomskega dela predstavlja razvoj programske opreme v okolju Microsoft Visual Studio 2013. Programska oprema zajema krmiljenje vhodno-izhodnih signalov preko vzporednih vrat ter 32-kanalnega digitalnega vhodno-izhodnega modula, sistem programskih prekinitev za obdelavo slik v realnem času, funkcije za sledenje kosov in algoritem za iskanje povezanih komponent, funkcije meritev ter grafični uporabniški vmesnik.The aim of this thesis is to present the development of a wood biscuit measuring and sorting system. A description of an automatic mechanical line consisting of a reservoir, an elevator, a vibrating rotary supplier, a measuring system and a sorting system that separates perfect wood biscuits from imperfect ones is included. The core part of the thesis is a presentation of software development in Microsoft Visual Studio 2013. The developed software includes controlling input/output signals via a parallel port and a 32-channel digital input/output module, a system of software interruptions for real time picture processing, functions that allow tracing of individual items, an algorithm for connected-component labelling, measuring functions and a graphic user interface

Guidelines on the management of valvular heart disease: The Task Force on the Management of Valvular Heart Disease of the European Society of Cardiology.

80These guidelines focus on valvular heart disease in adults and adolescents, are oriented towards management, and will not deal with endocarditis and congenital valve diseases in adults and adolescents, since recent guidelines have been produced by the ESC on these topics. Although valvular heart disease is less common in industrialized countries than coronary disease, heart failure, or hypertension, guidelines are needed in this field for several reasons: valvular heart disease is common and often requires intervention; substantial advances have been made in the understanding of its pathophysiology; the patient population has changed with a continuous decline of acute rheumatic fever and an increased incidence of degenerative valvular diseases in industrialized countries. The incidence of endocarditis remains stable and other causes of valve disease are rare. Because of the predominance of degenerative valve disease, the two most frequent valve diseases are now calcific aortic stenosis and mitral regurgitation. Aortic regurgitation and mitral stenosis have become less common. Diagnosis is now dominated by echocardiography, which has become the standard to evaluate valve structure and function. Treatment has not only developed through the continuing progress in prosthetic valve technology, but has also been reoriented by the development of conservative surgical approaches and the introduction of percutaneous interventional techniques.openopenVahanian, A; Baumgartner, ; H, ; Bax, ; J, ; Butchart, ; E, ; Dion, ; R, ; Filippatos, ; G, ; Flachskampf, ; F, ; Hall, ; R, ; Iung, ; B, ; Kasprzak, ; J, ; Nataf, ; P, ; Tornos, ; P, ; Torracca, ; L, ; Wenink, ; A, ; Silvia, ; Priori, G.; Blanc, Jean-Jacques; Andrzej, ; Budaj, ; John, ; Camm, ; Veronica, ; Dean, ; Jaap, ; Deckers, ; Kenneth, ; Dickstein, ; John, ; Lekakis, ; Keith, ; Mcgregor, ; Marco, ; Metra, ; João, ; Morais, ; Ady, ; Osterspey, ; Juan, ; Tamargo, ; Luis, José; Zamorano, ; Annalisa, ; Angelini, ; Manuel, ; Antunes, ; Angel, Miguel; Fernandez, Garcia; Christa, ; Gohlke-Baerwolf, ; Gilbert, ; Habib, ; John, ; Mcmurray, ; Catherine, ; Otto, ; Luc, ; Pierard, ; Josè, ; Pomar, L.; Bernard, ; Prendergast, ; Raphael, ; Rosenhek, ; Sousa, Miguel; Uva, ; Juan, ; Tamargo,Vahanian, A; Baumgartner, ; H, ; Bax, ; J, ; Butchart, ; E, ; Dion, ; R, ; Filippatos, ; G, ; Flachskampf, ; F, ; Hall, ; R, ; Iung, ; B, ; Kasprzak, ; J, ; Nataf, ; P, ; Tornos, ; P, ; Torracca, ; L, ; Wenink, ; A, ; Silvia, ; Priori, G.; Blanc, Jean Jacques; Andrzej, ; Budaj, ; John, ; Camm, ; Veronica, ; Dean, ; Jaap, ; Deckers, ; Kenneth, ; Dickstein, ; John, ; Lekakis, ; Keith, ; Mcgregor, ; Marco, ; Metra, Marco; João, ; Morais, ; Ady, ; Osterspey, ; Juan, ; Tamargo, ; Luis, José; Zamorano, ; Annalisa, ; Angelini, ; Manuel, ; Antunes, ; Angel, Miguel; Fernandez, Garcia; Christa, ; Gohlke, Baerwolf; Gilbert, ; Habib, ; John, ; Mcmurray, ; Catherine, ; Otto, ; Luc, ; Pierard, ; Josè, ; Pomar, L.; Bernard, ; Prendergast, ; Raphael, ; Rosenhek, ; Sousa, Miguel; Uva, ; Juan, ; Tamargo