Experimental investigation of the influence of supply temperature and supply pressure on the performance of a two axial groove hydrodynamic journal bearing

An experimental study of the influence of oil supply temperature and supply pressure on the performance of a 100mm plain journal bearing with two axial grooves located at ±90º to the load line was carried out. The hydrodynamic pressure at the mid-plane of the bearing, temperature profiles at the oil-bush and oil-shaft interfaces, bush torque, oil flow rate, and the position of the shaft were measured for variable operating conditions. Shaft rotational speed ranged from 1000 to 4000rpm and two different values of applied load were tested (2kN and 10kN). The supply temperature ranged from 35 to 50ºC, whereas the oil supply pressure range was 70kPa to 210kPa. Bearing performance is strongly dependent on the supply conditions. It was found that the existence of the downstream groove significantly affects the temperature profile at the oil-bush interface except for the low load, low feeding pressure cases, where the cooling effect of the upstream groove is significant. Feeding temperature has a strong effect on the minimum film thickness. The increase in maximum temperature is significantly lower than the corresponding increase in supply temperature. Increases in supply pressure lead to a significant rise in oil flow rate but have little effect on the maximum temperature and power-loss, except in the case of the lightly-loaded bearing. Shaft temperature was found to be close to the bearing maximum temperature for low applied loads, being significantly smaller than this value for high loads. The mean shaft temperature is only significantly higher than the outlet temperature at high shaft speeds

La propagation du projet de classe au sein de l'école primaire : une question d'influences sociales entre enseignants ?

The school project is based on official documents as is officially considered the guideline that should motivate the teaching staff. Research (Dubet, 1992; Rich, 1998, 2010; Gather Thurler, 2000) has shown that school projects, even if they are official, do not always become reality. This thesis proposes to take the perspective of classroom practice and not to begin with what the official texts require. Might not the process of dynamism within schools take its roots in the dissemination of class projects toward other classes or even the entire school? The phenomena of dissemination, if they indeed exist, are studied through the theories of social influence (Moscovici, 1979, 1984, and Mugny Perez, 1993). Can class projects influence other classes and even the entire school? This is the main question addressed in this work. The methodology of the study includes a preliminary investigation from eight exploratoryinterviews and a thorough quantitative phase with 166 questionnaires followed by another qualitative phase composed of 20 interviews. By crossing the data, the results point to a phenomenon of influence. Types of teacher behavior are also revealed through these mechanisms. This work analyzes these types of behaviors and the functioning of specialgroups in some schools.Le projet d’école est issu des textes officiels et constitue le projet institué qui doit animer les équipes pédagogiques. Des travaux (Dubet, 1992 ; Rich, 1998, 2010 ; Gather Thurler, 2000) montrent que les projets d’écoles, même s’ils sont écrits, ne prennent pas toujours vie dans la réalité des écoles. Cette thèse propose de se placer du côté des pratiques de classe et non du point de vue des textes. Des processus moteurs pour l’école ne pourraient-ils pas se trouver dans la propagation de projets de classe à d’autres classes, voire à l’école toute entière ? Les phénomènes de propagation, s’il y en a, sont étudiés à l’appui des théories de l’influence sociale (Moscovici, 1979, 1984 ; Perez et Mugny, 1993). Les projets de classe peuvent-ils influencer d’autres classes jusqu’à l’école ? Telle est la principale question posée. La méthodologie de l’étude comporte une enquête préalable à partir de huit entretiens exploratoires, puis une phase approfondie de type quantitatif avec 166 questionnaires suivie d’une autre phase approfondie de type qualitatif comportant 20 entretiens. Par croisement des données, les résultats mettent à jour des phénomènes d’influence. Des types de comportement d’enseignants émergent à l’occasion de ces mécanismes. Ce travail conduit à analyser ces types de comportements ainsi que des fonctionnements particuliers dans certaines écoles

The role of lubricant feeding conditions on the performance improvement and friction reduction of journal bearings

Most conventional hydrodynamic journal bearing performance tools can not suitably assess the effect of lubricant feeding conditions on bearing performance, even though these conditions are known to affect important performance parameters such as eccentricity and powerloss. A thermohydrodynamic analysis suitable to deal with realistic feeding conditions has been proposed. Special attention was given to the treatment of phenomena taking place within grooves and their vicinity,as well as to the ruptured film region. The effec to flubricant feeding pressure and temperature, groove length ratio,width ratio and number (single/twin) on bearing performance has been analyzed for a broad range of conditions.It was found that a careful tuning of the feeding conditions may indeed improve bearing performance.FCT - POCTI/EME/39202/200

Tumor protein D54 defines a new class of intracellular transport vesicles

Transport of proteins and lipids from one membrane compartment to another is via intracellular vesicles. We investigated the function of tumor protein D54 (TPD54/TPD52L2) and found that TPD54 was involved in multiple membrane trafficking pathways: anterograde traffic, recycling, and Golgi integrity. To understand how TPD54 controls these diverse functions, we used an inducible method to reroute TPD54 to mitochondria. Surprisingly, this manipulation resulted in the capture of many small vesicles (30 nm diameter) at the mitochondrial surface. Super-resolution imaging confirmed the presence of similarly sized TPD54-positive structures under normal conditions. It appears that TPD54 defines a new class of transport vesicle, which we term intracellular nanovesicles (INVs). INVs meet three criteria for functionality. They contain specific cargo, they have certain R-SNAREs for fusion, and they are endowed with a variety of Rab GTPases (16 out of 43 tested). The molecular heterogeneity of INVs and the diverse functions of TPD54 suggest that INVs have various membrane origins and a number of destinations. We propose that INVs are a generic class of transport vesicle that transfer cargo between these varied locations

Antibody-free magnetic cell sorting of genetically modified primary human CD4+ T cells by one-step streptavidin affinity purification.

Existing methods for phenotypic selection of genetically modified mammalian cells suffer disadvantages of time, cost and scalability and, where antibodies are used to bind exogenous cell surface markers for magnetic selection, typically yield cells coated with antibody-antigen complexes and beads. To overcome these limitations we have developed a method termed Antibody-Free Magnetic Cell Sorting in which the 38 amino acid Streptavidin Binding Peptide (SBP) is displayed at the cell surface by the truncated Low Affinity Nerve Growth Receptor (LNGFRF) and used as an affinity tag for one-step selection with streptavidin-conjugated magnetic beads. Cells are released through competition with the naturally occurring vitamin biotin, free of either beads or antibody-antigen complexes and ready for culture or use in downstream applications. Antibody-Free Magnetic Cell Sorting is a rapid, cost-effective, scalable method of magnetic selection applicable to either viral transduction or transient transfection of cell lines or primary cells. We have optimised the system for enrichment of primary human CD4+ T cells expressing shRNAs and exogenous genes of interest to purities of >99%, and used it to isolate cells following Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 genome editing

Evaluation of factors influencing expression and extraction of recombinant bacteriophage endolysins in Escherichia coli

Background: Endolysins are peptidoglycan hydrolases with promising use as environment-friendly antibacterials mainly when used topically. However, in general, endolysin expression is hampered by its low solubility. Thus, a critical point in endolysin industrial production is optimizing their expression, including improvement of solubility and recovery from cell extracts. Results: We report the expression of two endolysins encoded in the genome of phages infecting Staphylococcus aureus. Expression was optimized through changes in the concentration of the inducer and growth temperature during the expression. Usually, only 30–40% of the total endolysin was recovered in the soluble fraction. Co-expression of molecular chaperones (DnaK, GroEL) or N-term fusion tags endowed with increased solubility (DsbC, Trx, Sumo) failed to improve that yield substantially. Inclusion of osmolytes (NaCl, CaCl2, mannitol, glycine betaine, glycerol and trehalose) or tensioactives (Triton X-100, Tween 20, Nonidet P-40, CHAPS, N-lauroylsarcosine) in the cell disruption system (in the absence of any molecular chaperone) gave meager improvements excepted by N-lauroylsarcosine which increased recovery to 54% of the total endolysin content. Conclusion: This is the first attempt to systematically analyze methods for increasing yields of recombinant endolysins. We herein show that neither solubility tags nor molecular chaperones co-expression are effective to that end, while induction temperature, (His)6-tag location and lysis buffer additives (e.g.N-lauroylsarcosine), are sensible strategies to obtain higher levels of soluble S. aureus endolysins.Fil: Balaban, Cecilia Lucía. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Laboratorio de Microbiología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Suárez, Cristian Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Laboratorio de Microbiología Molecular; ArgentinaFil: Boncompain, Carina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Laboratorio de Microbiología Molecular; ArgentinaFil: Peressutti Bacci, Natalia. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Laboratorio de Microbiología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Ceccarelli, Eduardo Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Morbidoni, Héctor Ricardo. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Laboratorio de Microbiología Molecular; Argentin

Bioinformatic Analysis of a Set of 14 Temperate Bacteriophages Isolated from Staphylococcus aureus Strains Highlights Their Massive Genetic Diversity

Epidemiology and virulence studies of Staphylococcus aureus showed that temperate bacteriophages are one of the most powerful drivers for its evolution not only because of their abundance but also because of the richness of their genetic payload. Here, we report the isolation, genome sequencing, and bioinformatic analysis of 14 bacteriophages induced from lysogenic S. aureus strains from human or veterinary (cattle) origin. The bacteriophages belonged to the Siphoviridae family; were of similar genome size (40 to 45 kbp); and fell into clusters B2, B3, B5, and B7 according to a recent clustering proposal. One of the phages, namely, vB_SauS_308, was the most unusual one, belonging to the sparsely populated subcluster B7 but showing differences in protein family contents compared with the rest of the members. This phage contains a type I endolysin (one catalytic domain and noncanonical cell wall domain [CBD]) and a host recognition module lacking receptor binding protein, cell wall hydrolase, and tail fiber proteins. This phage also lacked virulence genes, which is opposite to what has been reported for subcluster B6 and B7 members. None of six phages, taken as representatives of each of the four subclusters, showed activity on coagulase-negative staphylococci (excepted for two Staphylococcus hominis strains in which propagation and a very slow adsorption rate were observed) nor transducing ability. Immunity tests on S. aureus RN4220 lysogens with each of these phages showed no cross immunity.Fil: Suárez, Cristian Alejandro. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Carrasco, Soledad Telma. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Brandolisio, Facundo Nahuel Adrián. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Abatángelo, Virginia. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Boncompain, Carina Andrea. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Peressutti Bacci, Natalia. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Carrasco, Soledad Telma. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentin

The endosomal transcriptional regulator RNF11 integrates degradation and transport of EGFR

Stimulation of cells with epidermal growth factor (EGF) induces internalization and partial degradation of the EGF receptor (EGFR) by the endo-lysosomal pathway. For continuous cell functioning, EGFR plasma membrane levels are maintained by transporting newly synthesized EGFRs to the cell surface. The regulation of this process is largely unknown. In this study, we find that EGF stimulation specifically increases the transport efficiency of newly synthesized EGFRs from the endoplasmic reticulum to the plasma membrane. This coincides with an up-regulation of the inner coat protein complex II (COPII) components SEC23B, SEC24B, and SEC24D, which we show to be specifically required for EGFR transport. Up-regulation of these COPII components requires the transcriptional regulator RNF11, which localizes to early endosomes and appears additionally in the cell nucleus upon continuous EGF stimulation. Collectively, our work identifies a new regulatory mechanism that integrates the degradation and transport of EGFR in order to maintain its physiological levels at the plasma membrane

Caracterización fenotípica y molecular de aislamientos de Staphylococcus aureus portados en narinas de pacientes con psoriasis que concurren a un hospital público en Argentina

A pesar de que Staphylococcus aureus incrementa su abundancia relativa en la psoriasis cuando se compara con el microbioma de personas sanas, no es el microorganismo más importante subyacente a la enfermedad. Sin embargo, existen pocos datos sobre el papel y las características moleculares de las cepas de S. aureus en pacientes con psoriasis. Nuestro objetivo fue evaluar la portación nasal de este microorganismo, sus características fenotípicas y moleculares, y el impacto de factores del hospedador sobre dicha portación en estos pacientes. Se analizó la presencia de S. aureus en hisopados nasales de 46 voluntarios sanos y 50 pacientes con psoriasis mediante técnicas microbiológicas convencionales. Se encontró mayor portación en pacientes con psoriasis que en el grupo control (37,24% vs. 22,98%, respectivamente) y esta estuvo asociada al sexo (masculino), la edad (adultos) y la gravedad de la enfermedad (más frecuente en casos moderados a graves). El 12% de los aislamientos de S. aureus mostraron resistencia a betalactámicos, con resistencia acompanante ˜ a macrólidos, aminoglucósidos y fluoroquinolonas en grado variable. No se encontró resistencia a rifampicina, vancomicina, mupirocina o trimetroprima/sulfametoxazol. Se realizó una caracterización molecular preliminar de los aislamientos por amplificación de genes de virulencia mediante PCR. Si bien se identificaron factores relacionados con el hospedador que incrementan la portación nasal de S. aureus en pacientes con psoriasis, la caracterización molecular de las cepas no reveló ninguna característica genotípica predominante asociada a esta afección. Se necesitan más estudios genómicos y transcriptómicos para profundizar en esta caracterización.Although Staphylococcus aureus increases its relative abundance in psoriasis when compared with the microbiome of healthy subjects, it is not the most important microorganism underlying this disease. However, there is scant data on the role and molecular features of S. aureus strains in psoriasis; therefore, the aim of this study was to evaluate nasal carriage of this microorganism, its phenotypic and molecular characteristics as well as the impact of host factors on its carriage in psoriatic patients. The presence of S. aureus was analyzed in nasal swabs from 46 healthy volunteers and 50 psoriatic patients by conventional microbiology techniques. Nasal carriage of S. aureus was higher in psoriatic patients than in the control group (37.24% vs 22.98%, respectively), being associated to sex (male), age (adults) and severity of the disease (more frequent in moderate and severe cases). Determination of antibiotic resistance detected 12% of β-lactam resistant isolates, with variable accompanying resistance to macrolides, aminoglycosides and fluoroquinolones. No resistance to rifampicin, vancomycin, mupirocin or trimethoprim/sulfamethoxazole was found. A preliminary molecular characterization of the isolates was performed by PCR amplification of virulence genes. Molecular characterization of the strains did not reveal a predominant strain in psoriatic patients. Although we established host factors related to increased carriage of S. aureus in psoriatic patients, we could not establish the predominance of one type of strain. Genomic and transcriptomic analysis of the isolated strains would be necessary to address this point.Fil: Boncompain, Carina Andrea. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Laboratorio de Microbiología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Suárez, Cristian Alejandro. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Laboratorio de Microbiología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Squeff, Mario Alfredo. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Belluzo, Valeria. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Piccirilli, G.. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Molteni, Ana. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Fernández Bussy, Ramón A.. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Morbidoni, Héctor Ricardo. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Laboratorio de Microbiología Molecular; Argentin

Shigella Effector IpaB-Induced Cholesterol Relocation Disrupts the Golgi Complex and Recycling Network to Inhibit Host Cell Secretion

Shigella infection causes destruction of the human colonic epithelial barrier. The Golgi network and recycling endosomes are essential for maintaining epithelial barrier function. Here we show that Shigella epithelial invasion induces fragmentation of the Golgi complex with consequent inhibition of both secretion and retrograde transport in the infected host cell. Shigella induces tubulation of the Rab11-positive compartment, thereby affecting cell surface receptor recycling. The molecular process underlying the observed damage to the Golgi complex and receptor recycling is a massive redistribution of plasma membrane cholesterol to the sites of Shigella entry. IpaB, a virulence factor of Shigella that is known to bind cholesterol, is necessary and sufficient to induce Golgi fragmentation and reorganization of the recycling compartment. Shigella infection-induced Golgi disorganization was also observed in vivo, suggesting that this mechanism affecting the sorting of cell surface molecules likely contributes to host epithelial barrier disruption associated with Shigella pathogenesis