Supermacroporous Polymer-Based Cryogel Bioreactor for Monoclonal Antibody Production in Continuous Culture Using Hybridoma Cells

Cryogel matrices composed of different polymeric blends were synthesized, yielding a unique combination of hydrophilicity and hydrophobicity with the presence or absence of charged surface. Four such cryogel matrices composed of polyacrylamide-chitosan (PAAC), poly(N-isopropylacrylamide)-chitosan, polyacrylonitrile (PAN), and poly(N-isopropylacrylamide) were tested for growth of different hybridoma cell lines and production of antibody in static culture. All the matrices were capable for the adherence of hybridoma cell lines 6A4D7, B7B10, and H9E10 to the polymeric surfaces as well as for the efficient monoclonal antibody (mAb) production. PAAC proved to be relatively better in terms of both mAb production and cell growth. Further, PAAC cryogel was designed into three different formats, monolith, disks, and beads, and used as packing material for packed-bed bioreactor. Longterm cultivation of 6A4D7 cell line on PAAC cryogel scaffold in all the three formats could be successfully done for a period of 6 weeks under static conditions. Continuous packed-bed bioreactor was setup using 6A4D7 hybridoma cell line in the three reactor formats. The reactors ran continuously for a period of 60 days during which mAb production and metabolism of cells in the bioreactors were monitored periodically. The monolith bioreactor performed most efficiently over a period of 60 days and produced a total of 57.5 mg of antibody in the first 30 days (in 500 mL) with a highest concentration of 115 mu g mL(-1), which is fourfold higher than t-flask culture. The results demonstrate that appropriate chemistry and geometry of the bioreactor matrix for cell growth and immobilization can enhance the reactor productivity. (C) 2010 American Institute of Chemical Engineers Biotechnol. Prog., 27: 170-180, 2011

Geometry and quadratic nonlinearity of charge transfer complexes in solution using depolarized hyper-Rayleigh scattering

We report large quadratic nonlinearity in a series of 1:1 molecular complexes between methyl substituted benzene donors and quinone acceptors in solution. The first hyperpolarizability, beta(HRS), which is very small for the individual components, becomes large by intermolecular charge transfer (CT) interaction between the donor and the acceptor in the complex. In addition, we have investigated the geometry of these CT complexes in solution using polarization resolved hyper-Rayleigh scattering (HRS). Using linearly (electric field vector along X direction) and circularly polarized incident light, respectively, we have measured two macroscopic depolarization ratios D = I-2 omega,I-X,I-X/I-2 omega,I-Z,I-X and D' = I-2 omega,I-X,I-C/I-2 omega,I-Z,I-C in the laboratory fixed XYZ frame by detecting the second harmonic scattered light in a polarization resolved fashion. The experimentally obtained first hyperpolarizability, beta(HRS), and the value of macroscopic depolarization ratios, D and D', are then matched with the theoretically deduced values from single and double configuration interaction calculations performed using the Zerner's intermediate neglect of differential overlap self-consistent reaction field technique. In solution, since several geometries are possible, we have carried out calculations by rotating the acceptor moiety around three different axes keeping the donor molecule fixed at an optimized geometry. These rotations give us the theoretical beta(HRS), D and D' values as a function of the geometry of the complex. The calculated beta(HRS), D, and D' values that closely match with the experimental values, give the dominant equilibrium geometry in solution. All the CT complexes between methyl benzenes and chloranil or 1,2-dichloro-4,5-dicyano-p-benzoquinone investigated here are found to have a slipped parallel stacking of the donors and the acceptors. Furthermore, the geometries are staggered and in some pairs, a twist angle as high as 30 degrees is observed. Thus, we have demonstrated in this paper that the polarization resolved HRS technique along with theoretical calculations can unravel the geometry of CT complexes in solution. (C) 2011 American Institute of Physics. doi:10.1063/1.3514922]

Geometry and quadratic nonlinearity of charge transfer complexes in solution: A theoretical study

In this paper, we have computed the quadratic nonlinear optical (NLO) properties of a class of weak charge transfer (CT) complexes. These weak complexes are formed when the methyl substituted benzenes (donors) are added to strong acceptors like chloranil (CHL) or di-chloro-di-cyano benzoquinone (DDQ) in chloroform or in dichloromethane. The formation of such complexes is manifested by the presence of a broad absorption maximum in the visible range of the spectrum where neither the donor nor the acceptor absorbs. The appearance of this visible band is due to CT interactions, which result in strong NLO responses. We have employed the semiempirical intermediate neglect of differential overlap (INDO/S) Hamiltonian to calculate the energy levels of these CT complexes using single and double configuration interaction (SDCI). The solvent effects are taken into account by using the self-consistent reaction field (SCRF) scheme. The geometry of the complex is obtained by exploring different relative molecular geometries by rotating the acceptor with respect to the fixed donor about three different axes. The theoretical geometry that best fits the experimental energy gaps, beta(HRS) and macroscopic depolarization ratios is taken to be the most probable geometry of the complex. Our studies show that the most probable geometry of these complexes in solution is the parallel displaced structure with a significant twist in some cases. (C) 2011 American Institute of Physics. doi:10.1063/1.3526748]

Proposed visible emission line space solar coronagraph

The outer atmosphere of the sun called the corona has been observed during total solar eclipse for short periods (typically <6 min), from as early as the eighteenth century. In the recent past, space-based instruments have permitted us to study the corona uninterruptedly. In spite of these developments, the dynamic corona and its high temperature (1-2 million K) are yet to be Ally understood. It is conjectured that their dynamic nature and associated energetic events are possible reasons behind the high temperature. In order to study these in detail, a visible emission line space solar coronagraph is being proposed as a payload under the small-satellite programme of the Indian Space Research Organisation. The satellite is named as Aditya-1 and the scientific objectives of this payload are to study: (i) the existence of intensity oscillations for the study of wave-driven coronal heating; (ii) the dynamics and formation of coronal loops and temperature structure of the coronal features; (iii) the origin, cause and acceleration of coronal mass ejections (CMEs) and other solar active features, and (iv) coronal magnetic field topology and three-dimensional structures of CMEs using polarization information. The uniqueness of this payload compared to previously flown space instruments is as follows: (a) observations in the visible wavelength closer to the disk (down to 1.05 solar radii); (b) high time cadence capability (better than two-images per second), and (c) simultaneous observations of at least two spectral windows all the time and three spectral windows for short durations.

Lattice embedding and equilibrium geometry of metal-halogen chains in the two-band extended Hubbard model

Two-band extended Hubbard model studies show that the shift in optical gap of the metal-halogen (MX) chain upon embedding in a crystalline environment depends upon alternation in the site-diagonal electron-lattice interaction parameter (epsilon(M)) and the strength of electron-electron interactions at the metal site (U(M)). The equilibrium geometry studies on isolated chains show that the MX chains tend to distort for alternating epsilon(M) and small U(M) values.

Geometry of proline-containing alpha-helices in proteins

Crystal structure analysis of proline-containing alpha-helices in proteins has been carried out. High resolution crystal structures were selected from the Protein Data Bank. Apart from the standard internal parameters, some parameters which are specifically related to the bend in the helix due to proline have been developed and analyzed. Finally the position and nature of these helices and their interactions with the rest of the protein have been analyzed.

Rectilinear Path Following in 3D Space

This paper addresses the problem of determining an optimal (shortest) path in three dimensional space for a constant speed and turn-rate constrained aerial vehicle, that would enable the vehicle to converge to a rectilinear path, starting from any arbitrary initial position and orientation. Based on 3D geometry, we propose an optimal and also a suboptimal path planning approach. Unlike the existing numerical methods which are computationally intensive, this optimal geometrical method generates an optimal solution in lesser time. The suboptimal solution approach is comparatively more efficient and gives a solution that is very close to the optimal one. Due to its simplicity and low computational requirements this approach can be implemented on an aerial vehicle with constrained turn radius to reach a straight line with a prescribed orientation as required in several applications. But, if the distance between the initial point and the straight line to be followed along the vertical axis is high, then the generated path may not be flyable for an aerial vehicle with limited range of flight path angle and we resort to a numerical method for obtaining the optimal solution. The numerical method used here for simulation is based on multiple shooting and is found to be comparatively more efficient than other methods for solving such two point boundary value problem.

Azidothymidine induces apoptosis in mouse myeloma cell line Sp2/0

Azidothymidine (AZT), which has been extensively used as an antiviral agent in the treatment of AIDS, showed strong inhibition of growth of Sp2/0 cells in vitro. AZT-treated cells showed a decrease in viability in a dose-dependent manner. AZT specifically induced typical apoptotic cell death with DNA double-strand cleavage and subsequent formation of apoptotic bodies. The induction of DNA double-strand cleavage into the oligonucleosomal ladder by AZT was protected in the presence of thymidine or uridine. An increase in endonuclease activity from nuclear extract of AZT-treated cells was observed. The enzyme activity was found to be Ca2+- and Mg2+-dependent and was inhibited by zinc acetate. A marked enhancement of PARP activity was observed in AZT-treated cells. These observations show that AZT can trigger both morphological and biochemical changes typical of apoptosis in the mouse myeloma cell line Sp2/0.

Line clipping revisited: Two efficient algorithms based on simple geometric observations

Two new line clipping algorithms, the opposite-corner algorithm and the perpendicular-distance algorithm, that are based on simple geometric observations are presented. These algorithms do not require computation of outcodes nor do they depend on the parametric representations of the lines. It is shown that the opposite-corner algorithm perform consistently better than an algorithm due to Nicholl, Lee, and Nicholl which is claimed to be better than the classic algorithm due to Cohen-Sutherland and the more recent Liang-Barsky algorithm. The pseudo-code of the opposite-corner algorithm is provided in the Appendix.

Geometry of sand-bed channels with seepage

Literature suggests that apart from a significant percentage of water loss from seepage, downward seepage from the channel causes an increase in the mobility of channel bed materials and thus changes the channel stability. Consequently, regime conditions (which provide the relationship among hydraulic parameters) should also be revised by incorporating downward seepage as an additional parameter. In the present work, regime conditions for the prediction of channel geometry in alluvial channels affected by downward seepage have been formulated on the basis of experimental observations. (C) 2011 Elsevier B.V. All rights reserved.

Irreversibility line and the hierarchy of weak links in Bi2Sr2CaCu2O8+?

The irreversibility line for H?c in a single crystal specimen of Bi2Sr2CaCu2O8+? (Bi2212) has been determined via vanishing of hysteresis in isothermal dc magnetization measurements. The hysteresis loops (H?c) in Bi2212 appear to show signatures of two-component magnetic response in several temperature regions where the temperature dependence of irreversibility field charges sharply. It is proposed that the observed behavior may be a consequence of existence of weak links of varying strength in Bi2212

Plate-shaped transformation products in zirconium-base alloys

Plate-shaped products resulting from martensitic, diffusional, and mixed mode transformations in zirconium-base alloys are compared. in the present study. These alloys are particularly suitable for the comparison in view of the fact that the lattice correspondence between the parent beta (bcc) and the product alpha (hcp) or gamma-hydride (fct) phases are remarkably similar for different types of transformations. Crystallographic features such as orientation relations, habit planes, and interface structures associated with these transformations have been compared:, with a view toward examining whether the transformation mechanisms have characteristic imprints on these experimental observables. Martensites exhibiting dislocated lath, internally twinned plate, and self-accommodating three-plate cluster morphologies have been encountered in Zr-2.5Nb alloy. Habit planes corresponding to all these morphologies have been found to be consistent with the predictions based on the invariant plane strain (IFS) criterion. Different morphologies have been found to reflect the manner in which the neighboring martensite variants are assembled. Lattice-invariant shears (LISs) for all these cases have been identified to be either {10 (1) over bar 1}(alpha) ((1) over bar 123)(alpha) slip or twinning on (10 (1) over bar 1)(alpha) planes. Widmanstatten alpha precipitates, forming in a step-quenching treatment, have been shown to have a lath morphology, the alpha/beta interface being decorated with a periodic array of (c + a) dislocations at a spacing of 8 to 10 nm. The line vectors of these dislocations are nearly parallel to the invariant lines. The alpha precipitates, forming in the retained beta phase on aging, exhibit an internally twinned structure with a zigzag habit plane. Average habit planes for the morphologies have been found to lie near the {103}(beta) - {113}(beta) poles, which are close to the specific variant of the {112}(beta) plane, which transforms into a prismatic plane of the type {1 (1) over bar 00}(alpha). The crystallography of the formation of the gamma-hydride phase (fct) from both the alpha and beta phases is seen to match the IFS predictions. While the beta-gamma transformation can be treated approximately as a simple shear on the basal plane involving a change in the stacking sequence, the alpha-gamma transformation call be conceptually broken into a alpha --> beta transformation following the Burgers correspondence and the simple beta-gamma shear process. The active eutectoid decomposition in the Zr-Cu system, beta --> alpha + beta', has been described in terms of cooperative growth of the alpha phase from the beta phase through the Burgers correspondence and of the partially ordered beta' (structurally similar to the equilibrium Zr2Cu phase) through an ordering process. Similarities and differences in crystallographic features of these transformations have been discussed. and the importance of the invariant line vector in deciding the geometry of the corresponding habit planes has been pointed out.

The algebra and geometry of SU(3) matrices

We give an elementary treatment of the defining representation and Lie algebra of the three-dimensional unitary unimodular group SU(3). The geometrical properties of the Lie algebra, which is an eight dimensional real Linear vector space, are developed in an SU(3) covariant manner. The f and d symbols of SU(3) lead to two ways of 'multiplying' two vectors to produce a third, and several useful geometric and algebraic identities are derived. The axis-angle parametrization of SU(3) is developed as a generalization of that for SU(2), and the specifically new features are brought out. Application to the dynamics of three-level systems is outlined.

Off-line signature verification using genetically optimized weighted features

This paper is concerned with off-line signature verification. Four different types of pattern representation schemes have been implemented, viz., geometric features, moment-based representations, envelope characteristics and tree-structured Wavelet features. The individual feature components in a representation are weighed by their pattern characterization capability using Genetic Algorithms. The conclusions of the four subsystems teach depending on a representation scheme) are combined to form a final decision on the validity of signature. Threshold-based classifiers (including the traditional confidence-interval classifier), neighbourhood classifiers and their combinations were studied. Benefits of using forged signatures for training purposes have been assessed. Experimental results show that combination of the Feature-based classifiers increases verification accuracy. (C) 1999 Pattern Recognition Society. Published by Elsevier Science Ltd. All rights reserved.

Helical structures: The geometry of protein helices and nanotubes

In nature, helical structures arise when identical structural subunits combine sequentially, the orientational and translational relation between each unit and its predecessor remaining constant. A helical structure is thus generated by the repeated action of a screw transformation acting on a subunit. A plane hexagonal lattice wrapped round a cylinder provides a useful starting point for describing the helical conformations of protein molecules, for investigating the geometrical properties of carbon nanotubes, and for certain types of dense packings of equal spheres.