Search results for: PAAR
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 7

Search results for: PAAR

7 Manipulating The PAAR Proteins of Acinetobacter Baumannii

Authors: Irene Alevizos, Jessica Lewis, Marina Harper, John Boyce

Abstract:

Acinetobacter baumannii causes a range of severe nosocomial-acquired infections, and many strains are multi-drug resistant. A. baumannii possesses survival mechanisms allowing it to thrive in competitive polymicrobial environments, including a Type VI Secretion System (T6SS) that injects effector proteins into other bacteria to give a competitive advantage. The effects of T6SS firing are broad and depend entirely on the effector that is delivered. Effects can include toxicity against prokaryotic or eukaryotic cells and the acquisition of essential nutrients. The T6SS of some species can deliver ‘specialised effectors’ that are fused directly to T6SS components, such as PAAR proteins. PAAR proteins are predicted to form the piercing tip of the T6SS and are essential for T6SS function. Although no specialised effectors have been identified in A. baumannii, many strains encode multiple PAAR proteins. Analysis of PAAR proteins across the species identified 12 families of PAAR proteins with distinct C-terminal extensions. A. baumannii AB307-0294 encodes two PAAR proteins, one of which has a C-terminal extension. Mutation of one or both of the PAAR-encoding genes in this strain showed that expression of either PAAR protein was sufficient for T6SS function. We employed a heterologous expression approach and determined that PAAR proteins from different A. baumannii strains, as well as the closely related A. baylyi species, could complement the A. baumannii ∆paar mutant and restore T6SS function. Furthermore, we showed that PAAR fusions could be used to deliver artificially cloned protein fragments by generating Histidine- and Streptavidin- tagged PAAR specialised effectors, which restored T6SS activity. This provides evidence that the fusion of protein fragments onto PAAR proteins in A. baumannii is compatible with a functional T6SS. Successful delivery by this mechanism extends the scope of what the T6SS can deliver, including user designed proteins.

Keywords: A. baumannii, effectors, PAAR, T6SS

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6 High-Temperature X-Ray Powder Diffraction of Secondary Gypsum

Authors: D. Gazdič, I. Hájková, M. Fridrichová

Abstract:

This paper involved the performance of a high-temperature X-Ray powder diffraction analysis (XRD) of a sample of chemical gypsum generated in the production of titanium white; this gypsum originates by neutralizing highly acidic water with limestone suspension. Specifically, it was gypsum formed in the first stage of neutralization when the resulting material contains, apart from gypsum, a number of waste products resulting from the decomposition of ilmenite by sulphuric acid. So it can be described as red titanogypsum. By conducting the experiment using XRD apparatus Bruker D8 Advance with a Cu anode (λkα=1.54184 Å) equipped with high-temperature chamber Anton Paar HTK 16, it was possible to identify clearly in the sample each phase transition in the system of CaSO4•xH2O.

Keywords: anhydrite, gypsum, bassanite, hematite, XRD, powder, high-temperature

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5 Density Measurement of Mixed Refrigerants R32+R1234yf and R125+R290 from 0°C to 100°C and at Pressures up to 10 MPa

Authors: Xiaoci Li, Yonghua Huang, Hui Lin

Abstract:

Optimization of the concentration of components in mixed refrigerants leads to potential improvement of either thermodynamic cycle performance or safety performance of heat pumps and refrigerators. R32+R1234yf and R125+R290 are two promising binary mixed refrigerants for the application of heat pumps working in the cold areas. The p-ρ-T data of these mixtures are one of the fundamental and necessary properties for design and evaluation of the performance of the heat pumps. Although the property data of mixtures can be predicted by the mixing models based on the pure substances incorporated in programs such as the NIST database Refprop, direct property measurement will still be helpful to reveal the true state behaviors and verify the models. Densities of the mixtures of R32+R1234yf an d R125+R290 are measured by an Anton Paar U shape oscillating tube digital densimeter DMA-4500 in the range of temperatures from 0°C to 100 °C and pressures up to 10 MPa. The accuracy of the measurement reaches 0.00005 g/cm³. The experimental data are compared with the predictions by Refprop in the corresponding range of pressure and temperature.

Keywords: mixed refrigerant, density measurement, densimeter, thermodynamic property

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4 Borate Crosslinked Fracturing Fluids: Laboratory Determination of Rheology

Authors: Lalnuntluanga Hmar, Hardik Vyas

Abstract:

Hydraulic fracturing has become an essential procedure to break apart the rock and release the oil or gas which are trapped tightly in the rock by pumping fracturing fluids at high pressure down into the well. To open the fracture and to transport propping agent along the fracture, proper selection of fracturing fluids is the most crucial components in fracturing operations. Rheology properties of the fluids are usually considered the most important. Among various fracturing fluids, Borate crosslinked fluids have proved to be highly effective. Borate in the form of Boric Acid, borate ion is the most commonly use to crosslink the hydrated polymers and to produce very viscous gels that can stable at high temperature. Guar and HPG (Hydroxypropyl Guar) polymers are the most often used in these fluids. Borate gel rheology is known to be a function of polymer concentration, borate ion concentration, pH, and temperature. The crosslinking using Borate is a function of pH which means it can be formed or reversed simply by altering the pH of the fluid system. The fluid system was prepared by mixing base polymer with water at pH ranging between 8 to 11 and the optimum borate crosslinker efficiency was found to be pH of about 10. The rheology of laboratory prepared Borate crosslinked fracturing fluid was determined using Anton Paar Rheometer and Fann Viscometer. The viscosity was measured at high temperature ranging from 200ᵒF to 250ᵒF and pressures in order to partially stimulate the downhole condition. Rheological measurements reported that the crosslinking increases the viscosity, elasticity and thus fluid capability to transport propping agent.

Keywords: borate, crosslinker, Guar, Hydroxypropyl Guar (HPG), rheology

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3 From Liquid to Solid: Advanced Characterization of Glass Applying Oscillatory Rheometry

Authors: Christopher Giehl, Anja Allabar, Daniela Ehgartner

Abstract:

Rotational rheometry is standard practice for the viscosity measurement of molten glass, neglecting the viscoelastic properties of this material, especially at temperatures approaching the glass transition. Oscillatory rheometry serves as a powerful toolbox for glass melt characterization beyond viscosity measurements. Heating and cooling rates and the time-dependent visco-elastic behavior influence the temperature where materials undergo the glass transition. This study presents quantitative thermo-mechanical visco-elasticity measurements on three samples in the Na-K-Al-Si-O system. The measurements were performed with a Furnace Rheometer System combined with an air-bearing DSR 502 measuring head (Anton Paar) and a Pt90Rh10 measuring geometry. Temperature ramps were conducted in rotation and oscillation, and the (complex) viscosity values were compared to calculated viscosity values based on sample composition. Furthermore, temperature ramps with different frequencies were conducted, also revealing the frequency-dependence of the shear loss modulus G’’ and the shear storage modulus G’. Here, lower oscillatory frequency results in lower glass transition temperature, as defined by the G’-G’’ crossover point. This contribution demonstrates that oscillatory rheometry serves as a powerful toolbox beyond viscosity measurements, as it considers the visco-elasticity of glass melts quantifying viscous and elastic moduli. Further, it offers a strong definition of Tg beyond the 10^12 Pas concept, which cannot be utilized with rotational viscometry data.

Keywords: frequency dependent glass transition, Na-K-Al-Si-O glass melts, oscillatory rheometry, visco-elasticity

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2 Friction Stir Processing of the AA7075T7352 Aluminum Alloy Microstructures Mechanical Properties and Texture Characteristics

Authors: Roopchand Tandon, Zaheer Khan Yusufzai, R. Manna, R. K. Mandal

Abstract:

Present work describes microstructures, mechanical properties, and texture characteristics of the friction stir processed AA7075T7352 aluminum alloy. Phases were analyzed with the help of x-ray diffractometre (XRD), transmission electron microscope (TEM) along with the differential scanning calorimeter (DSC). Depth-wise microstructures and dislocation characteristics from the nugget-zone of the friction stir processed specimens were studied using the bright field (BF) and weak beam dark-field (WBDF) TEM micrographs, and variation in the microstructures as well as dislocation characteristics were the noteworthy features found. XRD analysis display changes in the chemistry as well as size of the phases in the nugget and heat affected zones (Nugget and HAZ). Whereas the base metal (BM) microstructures remain un-affected. High density dislocations were noticed in the nugget regions of the processed specimen, along with the formation of dislocation contours and tangles. .The ɳ’ and ɳ phases, along with the GP-Zones were completely dissolved and trapped by the dislocations. Such an observations got corroborated to the improved mechanical as well as stress corrosion cracking (SCC) performances. Bulk texture and residual stress measurements were done by the Panalytical Empyrean MRD system with Co- kα radiation. Nugget zone (NZ) display compressive residual stress as compared to thermo-mechanically(TM) and heat affected zones (HAZ). Typical f.c.c. deformation texture components (e.g. Copper, Brass, and Goss) were seen. Such a phenomenon is attributed to the enhanced hardening as well as other mechanical performance of the alloy. Mechanical characterizations were done using the tensile test and Anton Paar Instrumented Micro Hardness tester. Enhancement in the yield strength value is reported from the 89MPa to the 170MPa; on the other hand, highest hardness value was reported in the nugget-zone of the processed specimens.

Keywords: aluminum alloy, mechanical characterization, texture characterstics, friction stir processing

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1 Targetting T6SS of Klebsiella pneumoniae for Assessment of Immune Response in Mice for Therapeutic Lead Development

Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

Abstract:

Klebsiella pneumoniae bacteria is a global threat to human health due to an increase in multi-drug resistance among strains. The hypervirulent strains of Klebsiella pneumoniae is a major trouble due to their association with life-threatening infections in a healthy population. One of the major virulence factors of hyper virulent strains of Klebsiella pneumoniae is the T6SS (Type six secretary system) which is majorly involved in microbial antagonism and causes interaction with the host eukaryotic cells during infections. T6SS mediates some of the crucial factors for establishing infection by the bacteria, such as cell adherence, invasion, and subsequent in vivo colonisation. The antibacterial activity and the cell invasion property of the T6SS system is a major requirement for the establishment of K. pneumoniae infections within the gut. The T6SS can be an appropriate target for developing therapeutics. The T6SS consists of an inner tube comprising hexamers of Hcp (Haemolysin -regulated protein) protein, and at the top of this tube sits VgrG (Valine glycine repeat protein G); the tip of the machinery consists of PAAR domain containing proteins which act as a delivery system for bacterial effectors. For this study, immune response to recombinant VgrG protein was generated to establish this protein as a potential immunogen for the development of therapeutic leads. The immunogenicity of the selected protein was determined by predicting the B cell epitopes by the BCEP analysis tool. The gene sequence for multiple domains of VgrG protein (phage_base_V, T6SS_Vgr, DUF2345) was selected and cloned in pMAL vector in E. coli. The construct was subcloned and expressed as a fusion protein of 203 residue protein with mannose binding protein tag (MBP) to enhance solubility and purification of this protein. The purified recombinant VgrG fusion protein was used for mice immunisation. The antiserum showed reactivity with the recombinant VgrG in ELISA and western blot. The immunised mice were challenged with K. pneumoniae bacteria and showed bacterial clearance in immunised mice. The recombinant VgrG protein can further be used for studying downstream signalling of VgrG protein in mice during infection and for therapeutic MAb development to eradicate K. pneumoniae infections.

Keywords: immune response, Klebsiella pneumoniae, multi-drug resistance, recombinant protein expression, T6SS, VgrG

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