Cytokinins in developing fruits of Moringa pterigosperma Gaertn.

The existence of cytokinins both as a free form and as a constituent of t-RNA was investigated in young fruits of Moringa pterigosperma Gaertn. Purified methanol extract was separated into butanol insoluble and butanol soluble fractions. The cytokinin(s) in the butanol insoluble fraction was tentatively identified as zeatin nucleotide. The butanol soluble fraction contained cytokinins and was chromatographed on Sephadex LH-20 with 35% ethanol. The two active fractions from LH-20 column coincided with zeatin and zeatin riboside. Cytokinin per g tissue was high in early stages of fruit growth and then remained more or less constant. Alkaline phosphatase hydrolysis of t-RNA hydrolysate of fruit tissue showed considerable cytokinin activity.     

Analysis and modeling of the variable region of camelid single-domain antibodies

Camelids have a special type of Ab, known as heavy chain Abs, which are devoid of classical Ab light chains. Relative to classical Abs, camelid heavy chain Abs (cAbs) have comparable immunogenicity, Ag recognition diversity and binding affinities, higher stability and solubility, and better manufacturability, making them promising candidates for alternate therapeutic scaffolds. Rational engineering of cAbs to improve therapeutic function requires knowledge of the differences of sequence and structural features between cAbs and classical Abs. In this study, amino acid sequences of 27 cAb variable regions (V(H)H) were aligned with the respective regions of 54 classical Abs to detect amino acid differences, enabling automatic identification of cAb V(H)H CDRs. CDR analysis revealed that the H1 often (and sometimes the H2) adopts diverse conformations not classifiable by established canonical rules. Also, although the cAb H3 is much longer than classical H3 loops, it often contains common structural motifs and sometimes a disulfide bond to the H1. Leveraging these observations, we created a Monte Carlo-based cAb V(H)H structural modeling tool, where the CDR H1 and H2 loops exhibited a median root-mean-square deviation to natives of 3.1 and 1.5 ?, respectively. The protocol generated 8-12, 14-16, and 16-24 residue H3 loops with a median root-mean-square deviation to natives of 5.7, 4.5, and 6.8 ?, respectively. The large deviation of the predicted loops underscores the challenge in modeling such long loops. cAb V(H)H homology models can provide structural insights into interaction mechanisms to enable development of novel Abs for therapeutic and biotechnological use.     

Impact of flow on ligand-mediated bacterial flocculation

To understand the adhesion–fragmentation dynamics of bacterial aggregates (i.e., flocs), we model the aggregates as two ligand-covered rigid spheres. We develop and investigate a model for the attachment/detachment dynamics in a fluid subject to a homogeneous planar shear-flow. The binding ligands on the surface of the flocs experience attractive and repulsive surface forces in an ionic medium and exhibit finite resistance to rotation (via bond tilting). For certain range of material and fluid parameters, our results predict a nonlinear or hysteretic relationship between the binding/unbinding of the floc surface and the net floc velocity (translational plus rotational velocity). We show that the surface adhesion is promoted by increased fluid flow until a critical value, beyond which the bonds starts to yield. Moreover, adhesion is not promoted in a medium with low ionic strength, or flocs with bigger size or higher binder stiffness. The numerical simulations of floc-aggregate number density studies support these findings.     

Cytokinin profile during early phase of germination of gram seeds after application of benzylaminopurine

Qualitative and quantitative differences in cytokinin levels of gram seeds were observed during germination of control and 6-benzylaminopurine (BAP; 4.44 x 10^t-5 M) treated seeds. Zeatin riboside (ZR), zeatin (Z), dihydrozeatin (DHZ), isopentenyladenosine (iPA) and isopentenyladenine (iP) were determined using high pressure liquid chromatography. In the control seeds the ZR content increased markedly only after 24 h of germination. In BAP treated seeds, the increase in ZR content was stronger than in control seeds and it persisted during further germination. Z content increased during germination in both control and treated seeds. After 48 h of germination iP appeared in control and especially in treated seeds. Acknowledgements: The senior author is thankful to University Grants Commision, New Delhi for award of Research Associateship.     

Purification and characterization of a glycoprotein from the surface of Ascaridia galli

Employing papain as the enzyme and agarose bound Ricinus communis agglutinin as the affinity gel, a glycoprotein has been isolated and purified from the surface of Ascaridia galli. The glycoprotein shows an apparent molecular weight of 68 kilo daltons and contains fucose, galactose, rhamnose and glucosamine as sugar moieties. Only 2% of its entire molecule has been found to possess alpha-helical configuration.     

2-Aryl-N-acyl indole derivatives as liver X receptor (LXR) agonists

Structure-activity relationship studies on a series of Boc-indole derivatives as LXR agonists are described. Compound 1 was identified as an LXR agonist through structure-based virtual screening followed by high-throughput gene profiling. Replacement of the indan linker portion in 1 with an open-chain linker resulted in compounds with similar or improved in vitro potency and cellular functional activity. The Boc group at the N-1 position of the indole moiety can be replaced with a benzoyl group. The SAR studies led to the identification of compound 8, a potent LXRbeta agonist with an EC50 of 12 nM in the cofactor recruitment assay.     

Synthesis and structure-activity relationship of 4-(2-aryl-cyclopropylamino)-quinoline-3-carbonitriles as EGFR tyrosine kinase inhibitors

Synthesis and structure-activity relationship of a series of 4-(2-aryl-cyclopropylamino)-quinoline-3-carbonitrile derivatives as EGFR inhibitors is described. Compounds 29 and 30 showed potent in vitro inhibitory activity in the enzymatic assay as well as in the functional cellular assay. They are moderately selective against other types of tyrosine kinases.