Development of Flow Cytometric Protocol for Nuclear DNA Content Estimation and Determination of Chromosome Number in Pongamia pinnata L., a Valuable Biodiesel Plant

The potentiality of Pongamia pinnata L. as a sustainable source of feedstock for the biodiesel industry is dependent on an extensive knowledge of the genome structure of the plant. Flow cytometry, with propidium iodide (PI) as the DNA stain, was used to estimate the nuclear DNA content of P. pinnata, with respect to Zea mays ‘CE-777’ as standard. The internal and pseudo-internal standardization was followed on account of the inhibitory effect of secondary compounds on PI intercalation. The antioxidants (PVP-40 and β-mercaptoethanol) were added to the nuclear isolation buffer for the reduction of inhibitory effect of P. pinnata cytosol. Nuclear DNA content estimation was done for P. pinnata leaves from different altitudes (37–117 m height from sea level) of Assam. Flow cytometry analysis indicated that the nuclear DNA content of P. pinnata is 2.66 pg with predicted 1C value of 1,300 Mb using Z. mays as standard. Coefficient of variation in flow cytometric analysis was within the limit of 5 % indicating that the results were reliable. Somatic chromosome numbers were counted from root–tip cells and was found to be 2n?=?22 corresponding to the diploid level (x?=?11). A decreasing trend in the nuclear DNA content was observed for the species of different altitudes.     

A facile Garratt–Braverman cyclization route to intercalative DNA-binding bis-quinones

Bispropargyl ethers (both symmetrical and non-symmetrical) equipped with 1,4-dimethoxyaryl groups were synthesized. Under strongly basic conditions (KOBu^t/toluene/reflux), these ethers underwent Garratt–Braverman type cyclization to the tetramethoxy bi-aryl systems in high yields presumably via the bisallenes. The products could be successfully converted to the bis-quinones via CAN-mediated demethylation cum oxidation. This two-step protocol offers a simple route to bis-quinones, connected by C1–C2′ bonds, in good yields. Fluorescence based EB-displacement assay, CD spectroscopy and viscosity measurements confirmed the DNA-binding ability of the synthesized quinones via intercalation.     

Bestimmung der Geschwindigkeitskonstante der Bromierungsreaktion von Acetanilid mittels Titrationsmethode in diffuser Schicht an der rotierenden Ring-Scheiben-Elektrode

The kinetics of the bromination of acetanilide has been studied with the diffusion layer titration method. The results have been obtained using the apparatus which consists of the ring-disc electrode and an amperostat-potentiostat system. Dependence of the ring current on the disc current has been determined as a function of rotation speeds of the electrode, of the solution concentration and temperature. It has been shown the bromination reaction of acetanilide exhibits by first order kinetics. In that case Br₂ and Br₃ ^? are the brominating species. The rate of bromination changes with the concentration of the Br^? ions. This reaction rate depends on reactions of molecular bromine with acetanilide. For 0.033<[Br^?]<0.173M the rate constant changes in the following range: $$17530M^{ - 1} s^{ - 1}< k< 6400M^{ - 1} s^{ - 1} ([H + ] = 1,34;T = 298K)$$      

Stepwise O2-Induced Rearrangement and Disassembly of the [NiFe4(OH)(μ3-S)4] Active Site Cluster of CO Dehydrogenase

Ni,Fe-containing carbon monoxide dehydrogenases (CODHs) catalyze the reversible reduction of carbon dioxide to carbon monoxide. CODHs are found in anaerobic microorganisms and can rapidly lose their activity when exposed to air. What causes the loss of activity is unclear. In this study, we analyzed the time-dependent structural changes induced by the presence of air on the metal centers of CODH-II. We show that inactivation is a multistep process. In a reversible step, the open coordination site on the Ni ion is blocked by a Ni,Fe-bridging μ-sulfido or chlorido ligand. Blocking this open coordination site with a cyanide ligand stabilizes the cluster against O₂-induced decomposition, indicating that O₂ attacks at the Ni ion. In the subsequent irreversible phase, nickel is lost, the Fe ions rearrange and the sulfido ligands disappear. Our data are consistent with a reversible reductive reactivation mechanism to protect CODHs from transient over-oxidation.     

Analytical method development for exploring pharmacokinetic profile of ursolic acid in rat tissues by high-performance thin-layer chromatography

JPC – Journal of Planar Chromatography – Modern TLC - A simple and rapid high-performance thin-layer chromatographic (HPTLC) determination of ursolic acid (UA) in tissue is necessary...     

Interlocked multiplets in anisotropically scattering medium with exponential Planck function

The equation of transfer for interlocked multiplets in anisotropically scattering atmosphere with planetary phase function has been considered. The Planck function is considered as exponential function of optical depth. The diffusely reflected intensity and emergent intensity in nth approximation has been obtained. Attempt has been made to get exact diffusely reflected intensity and emergent intensity.     

Characterization of buffers for electrokinetic separations

Buffers used in electrophoresis and electrochromatography must have a relatively low ionic strength in order to minimize ohmic heating in the presence of an applied potential. Calculation of pH, ionic strength, and the van Slyke buffer capacity, β, is therefore important. This paper describes thea priori calculation of these parameters for tris buffer made up with either glycine (a zwitterion) or HCl. A quadratic expression for pH, valid over wide ranges, is obtained for both buffer systems. The calculated values of pH, ionic strength, and buffer capacity are shown to agree with experimental results as a function of tris, HCl, and glycine concentrations ranging from 1 to 50 mM. A new parameter, the electrokinetic buffer effectiveness factor, is introduced to characterize buffers being considered for use in electrokinetic systems such as electrochromatography, and is used to determine the appropriate composition ranges for the buffer components.