Functional analysis of the large periplasmic loop of the Escherichia coli K-12 WaaL O-antigen ligase

WaaL is a membrane enzyme implicated in ligating undecaprenyl-diphosphate (Und-PP)-linked O antigen to lipid A-core oligosaccharide. We determined the periplasmic location of a large (EL5) and small (EL4) adjacent loops in the Escherichia coli K-12 WaaL. Structural models of the EL5 from the K-12, R1 and R4 E. coli ligases were generated by molecular dynamics. Despite the poor amino acid sequence conservation among these proteins, the models afforded similar folds consisting of two pairs of almost perpendicular alpha-helices. One alpha-helix in each pair contributes a histidine and an arginine facing each other, which are highly conserved in WaaL homologues. Mutations in either residue rendered WaaL non-functional, since mutant proteins were unable to restore O antigen surface expression. Replacements of residues located away from the putative catalytic centre and non-conserved residues within the centre itself did not affect ligation. Furthermore, replacing a highly conserved arginine in EL4 with various amino acids inactivates WaaL function, but functionality reappears when the positive charge is restored by a replacement with lysine. These results lead us to propose that the conserved amino acids in the two adjacent periplasmic loops could interact with Und-PP, which is the common component in all WaaL substrates.

Extracellular Loop 2 of the Free Fatty Acid Receptor 2 Mediates Allosterism of a Phenylacetamide Ago-Allosteric Modulator

Allosteric agonists are powerful tools for exploring the pharmacology of closely related G protein-coupled receptors that have nonselective endogenous ligands, such as the short chain fatty acids at free fatty acid receptors 2 and 3 (FFA2/GPR43 and FFA3/GPR41, respectively). We explored the molecular mechanisms mediating the activity of 4-chloro-alpha-(1-methylethyl)-N-2-thiazolylbenzeneacetamide (4-CMTB), a recently described phenylacetamide allosteric agonist and allosteric modulator of endogenous ligand function at human FFA2, by combining our previous knowledge of the orthosteric binding site with targeted examination of 4-CMTB structure-activity relationships and mutagenesis and chimeric receptor generation. Here we show that 4-CMTB is a selective agonist for FFA2 that binds to a site distinct from the orthosteric site of the receptor. Ligand structure-activity relationship studies indicated that the N-thiazolyl amide is likely to provide hydrogen bond donor/acceptor interactions with the receptor. Substitution at Leu(173) or the exchange of the entire extracellular loop 2 of FFA2 with that of FFA3 was sufficient to reduce or ablate, respectively, allosteric communication between the endogenous and allosteric agonists. Thus, we conclude that extracellular loop 2 of human FFA2 is required for transduction of cooperative signaling between the orthosteric and an as-yet-undefined allosteric binding site of the FFA2 receptor that is occupied by 4-CMTB.

The Bright Side of Manufacturing–Remanufacturing Conflict in a Decentralised Closed-Loop Supply Chain

Researchers and managers broadly agree that original equipment manufacturers (OEMs), which have opportunities to produce both new and remanufactured products, are better off by centrally controlling their manufacturing and remanufacturing activities. Thus, OEMs should not remanufacture used products until the remanufacturing cost is sufficiently low to overcome the negative impact of new product cannibalisation. In this paper, we present a contrasting view of the manufacturing–remanufacturing conflict: OEMs sometimes benefit from the decentralised control mode under which they ignore the internal cannibalisation rather than the remanufacturing option. We consider a decentralised closed-loop supply chain in which one OEM can purchase new components from one supplier to produce new products and collect used products from consumers to produce remanufactured products. The key feature of our model is that the OEM can select a centralised or decentralised control mode to manage its manufacturing and remanufacturing activities before the supplier prices the new component. In a steady state period setting, we analyse the players’ optimal decisions and compare the OEM's profits under centralised and decentralised control modes. Our analytic results reveal that the decentralised control within the OEM can outperform the centralised control when the cost structure of producing new and remanufactured products satisfies certain conditions. Finally, the key findings are distilled in a conceptual framework and its managerial implications are discussed.

Real-space mapping of dynamic phenomena during hysteresis loop measurements: Dynamic switching spectroscopy piezoresponse force microscopy

Dynamic switching spectroscopy piezoresponse force microscopy is developed to separate thermodynamic and kinetic effects in local bias-induced phase transitions. The approaches for visualization and analysis of five-dimensional data are discussed. The spatial and voltage variability of relaxation behavior of the a-c domain lead zirconate-titanate surface suggest the interpretation in terms of surface charge dynamics. This approach is applicable to local studies of dynamic behavior in any system with reversible bias-induced phase transitions ranging from ferroelectrics and multiferroics to ionic systems such as batteries, fuel cells, and electroresistive materials. (C) 2011 American Institute of Physics. [doi:10.1063/1.3590919]

Long-term graft survival in patients with flexible open-loop anterior chamber intraocular lenses

Purpose. To evaluate the long-term graft survival in patients with flexible open-loop anterior chamber intraocular lenses (AC IOL). Methods. We retrospectively reviewed the records of patients with aphakic/pseudophakic bullous keratopathy who underwent penetrating keratoplasty and flexible open-loop AC IOL implantation in our institution from 1983 to 1988. Results. 79 eyes from 77 patients were included in the study. Mean follow-up was 50 months (range 1 to 123 months). At last follow-up 61 eyes (77.2%) had clear grafts. Among them, the visual acuity was = 20/40 in 14 eyes (23.0%), 20/50-20/100 in 22 eyes (36.1%), 20/200-20/400 in 9 eyes (14.8%) and = CF in 16 (26.2%). Increment of glaucoma medications and/or glaucoma surgery was the most frequent complication (37 eyes, 46,8%). Cystoid macular edema was newly diagnosed in 10 eyes (12.7%). Conclusions. Flexible, open-loop anterior chamber lens are a viable option in the treatment of patients with aphakic or pseudophakic bullous keratopathy undergoing penetrating keratoplasty.

Long-term graft survival in patients with flexible open-loop anterior- chamber intraocular lenses

Purpose. To evaluate the long-term graft survival and complications of flexible, open-loop anterior-chamber intraocular lenses in patients with penetrating keratoplasty for pseudophakic or aphakic bullous keratopathy. Methods. We reviewed charts of all consecutive patients who underwent penetrating keratoplasty for pseudophakic or aphakic bullous keratopathy combined with implantation of a flexible, open-loop, anterior-chamber intraocular lens at our institution between 1983 and 1988. One-hundred one eyes of 99 patients were evaluated. Graft-survival rates were calculated by using the Kaplan-Meier actuarial method. Results. Mean follow-up was 49.8 months (range. 1-144). The probability of graft survival at 1, 2, 4, 6, and 8 years was 93, 87, 78, 65, and 65%, respectively. A total of 25 (24.8%) grafts failed. Progressive corneal edema without signs of rejection was the most common finding in patients with failed grafts (10 eyes, 40%). The most frequent complication observed was newly diagnosed or worsening of preexisting glaucoma (46 eyes, 45.5%). Conclusions. Our long-term results support flexible, open-loop anterior-chamber intraocular lenses as a reasonable option, at the time of penetrating keratoplasty, in patients with pseudophakic and aphakic bullous keratopathy.

SiGe V Band Wide Tuning-Range VCO and Frequency Divider for Phase Locked Loop

A V-band wide tuning-range VCO and high frequency divide-by-8 frequency divider using Infineon 0.35 µm SiGe HBT process are presented in this paper. An LC impedance peaking technique is introduced in the Miller divider to increase the sensitivity and operation frequency range of the frequency divider. Two static frequency dividers implemented using current mode logic are used to realize dividing by 4 in the circuit. The wide tuning range VCO operates from 51.9 to 64.1 GHz i.e. 20.3% frequency tuning range. The measured phase noise at the frequency divider output stage is around -98.5 dBc at 1 MHz. The circuit consumes 200mW and operates from a 3.5Vdc supply, and occupies 0.6×0.8 mm2 die area.

PLL with secondary loop control for self tracking antennas

This paper shows a simple, yet highly effective, tracking phase locked loop circuit which has applications for self steered antenna arrays. The tracking PLL has been demonstrated to accurately phase track signal levels as low as -120 dBm, making it suitable for applications such as SATCOM ground terminals. The implementation is simple requiring a low Q voltage controlled oscillator, a downconverting mixer and a PLL circuit.

Distributed closed-loop spatial multiplexing for uplink multiuser systems

Focusing on the uplink, where mobile users (each with a single transmit antenna) communicate with a base station with multiple antennas, we treat multiple users as antennas to enable spatial multiplexing across users. Introducing distributed closed-loop spatial multiplexing with threshold-based user selection, we propose two uplink channel-assigning strategies with limited feedback. We prove that the proposed system also outperforms the standard greedy scheme with respect to the degree of fairness, measured by the variance of the time averaged throughput. For uplink multi-antenna systems, we show that the proposed scheduling is a better choice than the greedy scheme in terms of the average BER, feedback complexity, and fairness. The numerical results corroborate our findings

Threshold-based substream selection for closed-loop spatial multiplexing

We propose a low-complexity closed-loop spatial multiplexing method with limited feedback over multi-input-multi-output (MIMO) fading channels. The transmit adaptation is simply performed by selecting transmit antennas (or substreams) by comparing their signal-to-noise ratios to a given threshold with a fixed nonadaptive constellation and fixed transmit power per substream. We analyze the performance of the proposed system by deriving closed-form expressions for spectral efficiency, average transmit power, and bit error rate (BER). Depending on practical system design constraints, the threshold is chosen to maximize the spectral efficiency (or minimize the average BER) subject to average transmit power and average BER (or spectral efficiency) constraints, respectively. We present numerical and Monte Carlo simulation results that validate our analysis. Compared to open-loop spatial multiplexing and other approaches that select the best antenna subset in spatial multiplexing, the numerical results illustrate that the proposed technique obtains significant power gains for the same BER and spectral efficiency. We also provide numerical results that show improvement over rate-adaptive orthogonal space-time block coding, which requires highly complex constellation adaptation. We analyze the impact of feedback delay using analytical and Monte Carlo approaches. The proposed approach is arguably the simplest possible adaptive spatial multiplexing system from an implementation point of view. However, our approach and analysis can be extended to other systems using multiple constellations and power levels.

TRACE observations of driven loop oscillations

Aims: On 13 June 1998, the TRACE satellite was fortuitously well placed to observe the effects of a flare-induced EIT wave in the corona, and its subsequent interaction with coronal magnetic loops. In this study, we use these TRACE observations to corroborate previous theoretical work, which determined the response of a coronal loop to a harmonic driver in the context of ideal magnetohydrodynamics, as well as estimate the magnetic field strength and the degree of longitudinal inhomogeneity. Methods: Loop edges are tracked, both spatially and temporally, using wavelet modulus maxima algorithms, with corresponding loop displacements from its quiescent state analysed by fitting scaled sinusoidal functions. The physical parameters of the coronal loop are subsequently determined using seismological techniques. Results: The studied coronal loop is found to oscillate with two distinct periods, 501 ± 5 s and 274 ± 7 s, which could be interpreted as belonging to the fundamental kink mode and first harmonic, or could reflect the stage of an overdriven loop. Additional scenarios for explaining the two periods are listed, each resulting in a different value of the magnetic field and the intrinsic and sub-resolution properties of the coronal loop. When assuming the periods belong to the fundamental kink mode and its first harmonic, we obtain a magnetic field strength inside the oscillating coronal loop of 2.0 ± 0.7 G. In contrast, interpreting the oscillations as a combination of the loop's natural kink frequency and a harmonic EIT wave provides a magnetic field strength of 5.8 ± 1.5 G. Using the ratio of the two periods, we find that the gravitational scale height in the loop is 73 ± 3 Mm. Conclusions: We show that the observation of two distinct periods in a coronal loop does not necessarily lead to a unique conclusion. Multiple plausible scenarios exist, suggesting that both the derived strength of the magnetic field and the sub-resolution properties of the coronal loop depend entirely on which interpretation is chosen. The interpretation of the observations in terms of a combination of the natural kink mode of the coronal loop, driven by a harmonic EIT wave seems to result in values of the magnetic field consistent with previous findings. Other interpretations, which are realistic, such as kink fundamental mode/first harmonic and the oscillations of two sub-resolution threads result in magnetic field strengths that are below the average values found before.