Facile preparation and some applications of an affinity matrix with a cleavable connector arm containing a disulfide bond

A versatile affinity matrix in which the ligand of interest is linked to the matrix through a connector arm containing a disulfide bond is described. It can be synthesized from any amino-substituted matrix by successive reaction with 2-imino-thio-lane, 5, 5'-dithiobis(2-nitrobenzoic acid), and a thiol derivative of the ligand of choice. The repertoire of ligands can be significantly increased by the appropriate use of avidin-biotin bridges. After adsorption of the material to be fractionated, elution can be effected by reducing the disulfide bond in the connector arm with dithiothreitol. Examples of the preparation and use of various affinity matrices based on amino-substituted Sepharose 6MB are given. One involves the immobilization of the Fab' fragment of a monoclonal antibody against Aspergillus oryzae β-galactosidase and the specific binding of that enzyme to the resulting immunoaffinity matrix. Another involves the immobilization of N-biotinyl-2-thioethylamine followed by complex formation with avidin. The resulting avidin-substituted matrix was used for the selective adsorption and subsequent recovery of mouse hybridoma cells producing anti-avidin antibodies. By further complexing the avidin-substituted matrix with appropriate biotinylated antigens, it should be possible to fractionate cells producing antibodies against a variety of antigens.

Hexa-arm star shaped hydrazone derivatives from hexakis(4-formylphenoxy)-cyclotriphosphazene core

A series of novel hexasubstituted cyclophosphazene hydrazones [N(3)P(3)(-OC(6)H(4)-p-CH=N-NH-C(O)-C(6)H(4)-p-X)(6)] (X = H, Br, Cl, F, OH, OCH(3), CH(3), NO(2), NH(2)) were prepared by a sixfold condensation reaction of [N(3)P(3)(-OC(6)H(4)-p-CHO)(6)] with para-substituted benzoic hydrazides [NH(2)-NH-C(O)-C(6)H(4)-p-X] with excellent yields (91-98%). The structures of the compounds were confirmed by elemental analysis, FT-IR, (1)H, (13)C, (31)P, 2D-HSQC NMR and mass spectrometry (MALDI-TOF). All the synthesized cyclophosphazene hydrazones exhibit high thermal stability. The crystal structure of a homogeneously substituted hexakis(4-formylphenoxy)-cyclotriphosphazene was determined by X-ray diffraction analysis. The compound crystallizes in the monoclinic system, space group P2(1)/n with a = 16.558(3) angstrom, b = 10.250(2) angstrom, c = 23.429(5) angstrom, alpha = gamma = 90.00 degrees, beta = 90.461(4)degrees, V = 3976.5(14) angstrom(3) and Z = 4. The R value is 0.0823 for 4290 observed reflections. The conformations of the 4-formylphenoxy-groups are different at the three phosphorus atoms. (C) 2011 Elsevier B.V. All rights reserved.

Whole arm manipulation using closed and hybrid planar chains: A kinematic study

This paper studies planar whole arm manipulation of a circular object using closed loop and hybrid manipulators. The manipulation is simple with fewer degrees of actuation than the task space. This is an useful operation if the initial and final positions of the object are on the same surface. Closed loop manipulator is a 4/5 bar mechanism. In hybrid manipulators a open loop manipulator with 3/4 links is attached to the floating link of 4/5 bar mechanism. The mobility analysis is carried out to find the connectivity of the object with reference to frame. The manipulation (forward kinematics) starts from a given configuration of the object and the manipulator. In hybrid manipulators determination of initial configuration involves inverse kinematics of open loop manipulator. The input joint velocities are used to demonstrate the manipulation. Conditions are specified for prehensile manipulation.

Magnetic arm-switch-based three-phase series-shunt compensated quality AC power supply

This study presents a novel magnetic arm-switch-based integrated magnetic circuit for a three-phase series-shunt compensated uninterruptible power supply (UPS). The magnetic circuit acts as a common interacting field for a number of energy ports, viz., series inverter, shunt inverter, grid and load. The magnetic arm-switching technique ensures equivalent series or shunt connection between the inverters. In normal grid mode (stabiliser mode), the series inverter is used for series voltage correction and the shunt one for current correction. The inverters and the load are effectively connected in parallel when the grid power is not available. These inverters are then used to share the load power. The operation of the inverters in parallel is ensured by the magnetic arm-switching technique. This study also includes modelling of the magnetic circuit. A graphical technique called bond graph is used to model the system. In this model, the magnetic circuit is represented in terms of gyrator-capacitors. Therefore the model is also termed as gyrator-capacitor model. The model is used to extract the dynamic equations that are used to simulate the system using MATLAB/SIMULINK. This study also discusses a synchronously rotating reference frame-based control technique that is used for the control of the series and shunt inverters in different operating modes. Finally, the gyrator-capacitor model is validated by comparing the simulated and experimental results.

An Implementation of Partitioned Scheduling Scheme for Hard Real-Time Tasks in Multicore Linux with Fair Share for Linux Tasks

The correctness of a hard real-time system depends its ability to meet all its deadlines. Existing real-time systems use either a pure real-time scheduler or a real-time scheduler embedded as a real-time scheduling class in the scheduler of an operating system (OS). Existing implementations of schedulers in multicore systems that support real-time and non-real-time tasks, permit the execution of non-real-time tasks in all the cores with priorities lower than those of real-time tasks, but interrupts and softirqs associated with these non-real-time tasks can execute in any core with priorities higher than those of real-time tasks. As a result, the execution overhead of real-time tasks is quite large in these systems, which, in turn, affects their runtime. In order that the hard real-time tasks can be executed in such systems with minimal interference from other Linux tasks, we propose, in this paper, an integrated scheduler architecture, called SchedISA, which aims to considerably reduce the execution overhead of real-time tasks in these systems. In order to test the efficacy of the proposed scheduler, we implemented partitioned earliest deadline first (P-EDF) scheduling algorithm in SchedISA on Linux kernel, version 3.8, and conducted experiments on Intel core i7 processor with eight logical cores. We compared the execution overhead of real-time tasks in the above implementation of SchedISA with that in SCHED_DEADLINE's P-EDF implementation, which concurrently executes real-time and non-real-time tasks in Linux OS in all the cores. The experimental results show that the execution overhead of real-time tasks in the above implementation of SchedISA is considerably less than that in SCHED_DEADLINE. We believe that, with further refinement of SchedISA, the execution overhead of real-time tasks in SchedISA can be reduced to a predictable maximum, making it suitable for scheduling hard real-time tasks without affecting the CPU share of Linux tasks.

Importance of amino-terminus in maintenance of oligomeric structure of cytosolic serine hydroxymethyltransferase

The role of the amino and carboxyl-terminal regions of cytosolic serine hydroxymethyltransferase (SHMT) in subunit assembly and catalysis was studied using six amino-terminal (lacking the first 6, 14, 30, 49, 58, and 75 residues) and two carboxyl-terminal (lacking the last 49 and 185 residues) deletion mutants. These mutants were constructed from a full length cDNA clone using restriction enzyme/PCR-based methods and overexpressed in Escherichia coli. The overexpressed proteins, des-(A1-K6)-SHMT and des-(A1- W14)-SHMT were present in the soluble fraction and they were purified to homogeneity. The deletion clones, for des-(A1–V30)-SHMT and des-(A1–L49)-SHMT were expressed at very low levels, whereas des-(A1–R58)-SHMT, des-(A1–G75)-SHMT, des-(Q435–F483)-SHMT and des-(L299-F483)-SHMT mutant proteins were not soluble and formed inclusion bodies. Des-(A1–K6)-SHMT and des-(A1–W14)-SHMT catalyzed both the tetrahydrofolate-dependent and tetrahydrofolate-independent reactions, generating characteristic spectral intermediates with glycine and tetrahydrofolate. The two mutants had similar kinetic parameters to that of the recombinant SHMT (rSHMT). However, at 55 °C, the des-(A1–W14)-SHMT lost almost all the activity within 5 min, while at the same temperature rSHMT and des-(A1–K6)-SHMT retained 85% and 70% activity, respectively. Thermal denaturation studies showed that des-(A1–W14)-SHMT had a lower apparent melting temperature (52°C) compared to rSHMT (56°C) and des-(A1–K6)-SHMT (55 °C), suggesting that N-terminal deletion had resulted in a decrease in the thermal stability of the enzyme. Further, urea induced inactivation of the enzymes revealed that 50% inactivation occurred at a lower urea concentration (1.2 ± 0.1 M) in the case of des-(A1–W14)-SHMT compared to rSHMT (1.8 ±0.1 M) and des-(A1–K6)-SHMT (1.7 ±0.1 M). The apoenzyme of des-(A1- W14)-SHMT was present predominantly in the dimer form, whereas the apoenzymes of rSHMT and des-(A1–K6)-SHMT were a mixture of tetramers (≈75% and ≈65%, respectively) and dimers. While, rSHMT and des-(A1–K6)-SHMT apoenzymes could be reconstituted upon the addition of pyridoxal-5'-phosphate to 96% and 94% enzyme activity, respectively, des-(A1–W14)-SHMT apoenzyme could be reconstituted only upto 22%. The percentage activity regained correlated with the appearance of visible CD at 425 nm and with the amount of enzyme present in the tetrameric form upon reconstitution as monitored by gel filtration. These results demonstrate that, in addition to the cofactor, the N-terminal arm plays an important role in stabilizing the tetrameric structure of SHMT.

Importance of the amino terminus in maintenance of oligomeric structure of sheep liver cytosolic serine hydroxymethyltransferase

The Role Of The Amino And Carboxyl-Terminal Regions Of Cytosolic Serine Hydroxymethyltransferase (SHMT) In Subunit Assembly And Catalysis Was Studied Using Sis Amino-Terminal (Lacking The First 6, 14, 30, 49, 58, And 75 Residues) And Two Carboxyl-Terminal (Lacking The Last 49 And 185 Residues) Deletion Mutants. These Mutants Were Constructed From A Full Length Cdna Clone Using Restriction Enzyme/PCR-Based Methods And Overexpressed In Escherichia Coli. The Overexpressed Proteins, Des-(A1-K6) SHMT And Des-(A1-W14)-SHMT Were Present In The Soluble Fraction And They Were Purified To Homogeneity. The Deletion Clones, For Des-(A1-V30)-SHMT And Des-(A1-L49)-SHMT Were Expressed At Very Low Levels, Whereas Des-(A1-R58)-SHMT, Des-/A1-G75)-SHMT, Des-(Q435-F483)-SHMT And Des-(L299-F483)-SHMT Mutant Proteins Were Not Soluble And Formed Inclusion Bodies. Des-(A1-K6)-SHMT And Des-(A1-W14)-SHMT Catalyzed Both The Tetrahydrofolate-Dependent And Tetrahydrofolate-Independent Reactions, Generating Characteristic Spectral Intermediates With Glycine And Tetrahydrofolate. The Two Mutants Had Similar Kinetic Parameters To That Of The Recombinant SHMT (Rshmt). However, At 55 Degrees C, The Des-(A1-W14)-SHMT Lost Almost All The Activity Within 5 Min, While At The Same Temperature Rshmt And Des-(A1-K6)-SHMT Retained 85% And 70% Activity, Respectively. Thermal Denaturation Studies Showed That Des-(A1-W14)-SHMT Had A Lower Apparent Melting Temperature (52 Degrees C) Compared To Rshmt (56 Degrees C) And Des-(A1-K6)-SHMT (55 Degrees C), Suggesting That N-Terminal Deletion Had Resulted In A Decrease In The Thermal Stability Of The Enzyme. Further Urea Induced Inactivation Of The Enzymes Revealed That 50% Inactivation Occurred At A Lower Urea Concentration (1.2+/-0.1 M) In The Case Of Des-(A1-W14)-SHMT Compared To Rshmt (1.8+/-0.1 M) And Des-(A1 -K6)-SHMT (1.7+/-0.1 M). The Apoenzyme Of Des-/A1-K6)-SHMT Was Present Predominantly In The Dimer Form, Whereas The Apoenzymes Of Rshmt And Des-(A1-K6)-SHMT Were A Mixture Of Tetramers (Approximate To 75% And Approximate To 65%, Respectively) And Dimers. While, Rshmt And Des-(A1-K6)-SHMT Apoenzymes Could Be Reconstituted Upon The Addition Of Pyridoxal-5'-Phosphate To 96% And 94% Enzyme Activity, Respectively Des-(A1-W14)-SHMT Apoenzyme Could Be Reconstituted Only Upto 22%. The Percentage Activity Refined Correlated With The Appearance Of Visible CD At 425 Nm And With The Amount Of Enzyme Present In The Tetrameric Form Upon Reconstitution As Monitored By Gel Filtration. These Results Demonstrate That, In Addition To The Cofactor, The N-Terminal Arm Plays An Important Role In Stabilizing The Tetrameric Structure Of SHMT.

A Scalable High Throughput Firewall in FPGA

High end network security applications demand high speed operation and large rule set support. Packet classification is the core functionality that demands high throughput in such applications. This paper proposes a packet classification architecture to meet such high throughput. We have implemented a Firewall with this architecture in reconflgurable hardware. We propose an extension to Distributed Crossproducting of Field Labels (DCFL) technique to achieve scalable and high performance architecture. The implemented Firewall takes advantage of inherent structure and redundancy of rule set by using our DCFL Extended (DCFLE) algorithm. The use of DCFLE algorithm results in both speed and area improvement when it is implemented in hardware. Although we restrict ourselves to standard 5-tuple matching, the architecture supports additional fields. High throughput classification invariably uses Ternary Content Addressable Memory (TCAM) for prefix matching, though TCAM fares poorly in terms of area and power efficiency. Use of TCAM for port range matching is expensive, as the range to prefix conversion results in large number of prefixes leading to storage inefficiency. Extended TCAM (ETCAM) is fast and the most storage efficient solution for range matching. We present for the first time a reconfigurable hardware implementation of ETCAM. We have implemented our Firewall as an embedded system on Virtex-II Pro FPGA based platform, running Linux with the packet classification in hardware. The Firewall was tested in real time with 1 Gbps Ethernet link and 128 sample rules. The packet classification hardware uses a quarter of logic resources and slightly over one third of memory resources of XC2VP30 FPGA. It achieves a maximum classification throughput of 50 million packet/s corresponding to 16 Gbps link rate for the worst case packet size. The Firewall rule update involves only memory re-initialization in software without any hardware change.

A do-it-yourself (DIY) switched mode power conversion laboratory

In the education of physical sciences, the role of the laboratory cannot be overemphasised. It is the laboratory exercises which enable the student to assimilate the theoretical basis, verify the same through bench-top experiments, and internalize the subject discipline to acquire mastery of the same. However the resources essential to put together such an environment is substantial. As a result, the students go through a curriculum which is wanting in this respect. This paper presents a low cost alternative to impart such an experience to the student aimed at the subject of switched mode power conversion. The resources are based on an open source circuit simulator (Sequel) developed at IIT Mumbai, and inexpensive construction kits developed at IISc Bangalore. The Sequel programme developed by IIT Mumbai, is a circuit simulation program under linux operating system distributed free of charge. The construction kits developed at IISc Bangalore, is fully documented for anyone to assemble these circuit which minimal equipment such as soldering iron, multimeter, power supply etc. This paper puts together a simple forward dc to dc converter as a vehicle to introduce the programming under sequel to evaluate the transient performance and small signal dynamic model of the same. Bench tests on the assembled construction kit may be done by the student for study of operation, transient performance and closed loop stability margins etc.

A Scalable High Throughput Firewall in FPGA

High end network security applications demand high speed operation and large rule set support. Packet classification is the core functionality that demands high throughput in such applications. This paper proposes a packet classification architecture to meet such high throughput. We have Implemented a Firewall with this architecture in reconfigurable hardware. We propose an extension to Distributed Crossproducting of Field Labels (DCFL) technique to achieve scalable and high performance architecture. The implemented Firewall takes advantage of inherent structure and redundancy of rule set by using, our DCFL Extended (DCFLE) algorithm. The use of DCFLE algorithm results In both speed and area Improvement when It is Implemented in hardware. Although we restrict ourselves to standard 5-tuple matching, the architecture supports additional fields.High throughput classification Invariably uses Ternary Content Addressable Memory (TCAM) for prefix matching, though TCAM fares poorly In terms of area and power efficiency. Use of TCAM for port range matching is expensive, as the range to prefix conversion results in large number of prefixes leading to storage inefficiency. Extended TCAM (ETCAM) is fast and the most storage efficient solution for range matching. We present for the first time a reconfigurable hardware Implementation of ETCAM. We have implemented our Firewall as an embedded system on Virtex-II Pro FPGA based platform, running Linux with the packet classification in hardware. The Firewall was tested in real time with 1 Gbps Ethernet link and 128 sample rules. The packet classification hardware uses a quarter of logic resources and slightly over one third of memory resources of XC2VP30 FPGA. It achieves a maximum classification throughput of 50 million packet/s corresponding to 16 Gbps link rate for file worst case packet size. The Firewall rule update Involves only memory re-initialiization in software without any hardware change.

Ionized Gas towards Galactic Centre-Constraints from Low-Frequency Recombination Lines

Observations of the H272α recombination line towards the galactic centre show features near VLSR= 0, -50 and + 36 kms-1 . We have combined the parameters of these features with the available -166 measurements to obtain the properties of the ionized gas present along the line of sight and also in the -3 kpc arm-. For the line-of-sight ionized gas we get an electron density around 7 cm-3 and a pathlength through it 10-60 pc. The emission measure and the electron temperature are in the range 500-2900 pc cm-6 and 2000-6000 - respectively. The ionized gas in the 3 kpc arm has an electron density of 30 cm-3 and extends over 9 pc along the line of sight if we assume an electron temperature of 104 K. Using the available upper limit to the intensity of the H351α recombination line, we show that the distributed ionized gas responsible for the dispersion of pulsar signals should have a temperature > 4500 - and a minimum filling factor of 20 per cent. We also show that recombination lines from the -warm ionized- gas proposed by McKee & Ostriker (1977) should be detectable in the frequency range 100-150 MHz towards the galactic centre with the sensitivity available at present.

Complex carbohydrates: 1. Conformational studies on some oligosaccharides related to N-glycosyl proteins which interact with concanavalin A

Empirical potential energy calculations have been carried out to determine the preferred conformations of some oligosaccharides having the trimannosidic core structure (Man3GlcNAc2) and which interact with concanavalin A. In the minimum energy conformations for the trimannosidic core the mannose residue on the Man α(1–6) arm comes close to one of the N-acetylglucosamine residues of the core. The addition of N-acetylglucosamine residues to the terminal mannose residues does not alter the preferred conformation of the trimannosidic core although it alters the relative preference of some of the higher energy conformations. The minimum energy conformation broadly agrees with available X-ray data. The presence of a bisecting N-acetylglucosamine residue on the middle mannose does not push the trimannosidic core to any new conformation but it does alter the relative preference for a particular conformation.

Microstrip square ring antenna for dual-band operation

This paper presents a generalized approach to design an electromagnetically coupled microstrip ring antenna for dual-band operation. By widening two opposite sides of a square ring antenna, its fractional bandwidth at the primary resonance mode can be increased significantly so that it may be used for practical applications. By attaching a stub to the inner edge of the side opposite to the feed arm, some of the losses in electrical length caused by widening can be regained. More importantly, this addition also alters the current distribution on the antenna and directs radiations at the second resonant frequency towards boresight. It has also been observed that for the dual frequency configurations studied, the ratio of the resonant frequencies (center dot r(2)center dot center dot r(1)) can range between 1.55 and 2.01. This shows flexibility in designing dual frequency antennas with a desired pair of resonant frequencies.