High Frequency Divisions in Leaf Base Protoplasts of Wheat (Triticum aestivum L.)

Protoplasts were isolated from the basal meristematic region of leaves from 6-day-old seedlings of wheat (Triticum aestivum). Protoplasts divided when cultured on MS medium (as agarose beads) in presence of nurse tissue. Donor seedlings when grown on BAP-supplemented MS medium were found to be considerably superior for protoplast isolation and culture than when grown on MS basal medium, in terms of protoplast viability, cell wall formation and cell division frequency. In addition, reduction of ammonium content of the culture medium, together with a dark Incubation, led to a high protoplast division frequency of about 70%. Microcolonies of 10-to 12-celled stages were obtained in Triticum aestivum, varieties HD 2329, HD 2285, Kalyan Sona, Arjun and CPAN 1676.     

Induced Stress on Red Blood Cell Promotes Red Blood Cell-Endothelial Adhesion

Cell and Tissue Biology - Besides disease condition, very few stress stimulants were determined to provoke red blood cell (RBC) adhesion to endothelial cells (EC). However, the possible role of...     

Ribonuclease from cobra snake venom: purification by affinity chromatography and further characterization

A ribonuclease from cobra snake venom was isolated and purified to homogeneity using antibody-affinity chromatography, increasing the yield fourfold. The purified enzyme showed cytidylic acid specificity, as reported earlier. Further, the effects of temperature, pH, metal ions, inhibitors, and urea on the enzyme activity were studied. Snake venom RNase exhibited salt-dependent reversible association-dissociation behaviour. Immunological studies indicate that this enzyme shares one of the antigenic sites of RNase A. The partial N-terminal sequence of the enzyme showed considerable homology with phospholipases from snake venom; however, the enzyme itself did not show any phospholipase activity.     

Identification of novel indigenous Bacillus thuringiensis isolates

Internally Transcribed Spacers (ITS) characterization and distribution of Repetitive Extragenic Palindromic (REP) sequences were studied in the genome of 223 field isolates of Bacillus thuringiensis from Madurai, India. They were characterized by morphological, biochemical and molecular methods. One hundred and twenty four of a total 223 isolates fitted ITS characterization of B. thuringiensis varieties known. Significant genomic variation was observed among seven isolates using REP primers. The ITS PCR product (EMBL accession number AJ639659) exhibited 98% nucleotide sequence homology with B. thuringiensis and placed the origin of indigenous isolate LDC-7 closer to B. thuringiensis on the basis of phylogenetic analysis.     

Biotransformation of Heavy Metals from Soil in Synthetic Medium Enriched with Glucose and Shewanella sp. HN-41 at Various pH

The biotransformation of heavy metals from contaminated soil was examined using a facultative anaerobic bacterium Shewanella sp. HN-41. The experiments were carried out to assess the influence of glucose at various pH on the transformation of heavy metals from soil thorough solubilization. A preliminary study on the transformation of heavy metals from soil was first performed using a defined medium supplemented with glucose at 10, 20, and 30 mM to select the effective concentration. Among the three concentrations examined, glucose at 30 mM leached a highest level of metal ions. Therefore, 30 mM glucose was used as the representative carbon source for the subsequent experiments in a defined medium at various pH (5, 6, 7, 8, and 9). The organism HN-41 was not influenced by pH ranging from acidic to neutral and was able to metabolize all the metal elements from contaminated soil. The level of Fe, Cr, As, Mn, Pb, and Al solubilization ranged from 3 to 7664 mg kg^?1 at various initial pH. The rate of metal solubilization was found to be low at neutral pH compared with acidic and alkaline. These results are expected to assist in the development of heavy metal transformation processes for the decontamination of heavy metal-contaminated soil.     

Influence of Protein – Micelle Ratios and Cysteine Residues on the Kinetic Stability and Unfolding Rates of Human Mitochondrial VDAC-2

Delineating the kinetic and thermodynamic factors which contribute to the stability of transmembrane β-barrels is critical to gain an in-depth understanding of membrane protein behavior. Human mitochondrial voltage-dependent anion channel isoform 2 (hVDAC-2), one of the key anti-apoptotic eukaryotic β-barrel proteins, is of paramount importance, owing to its indispensable role in cell survival. We demonstrate here that the stability of hVDAC-2 bears a strong kinetic contribution that is dependent on the absolute micellar concentration used for barrel folding. The refolding efficiency and ensuing stability is sensitive to the lipid-to-protein (LPR) ratio, and displays a non-linear relationship, with both low and high micellar amounts being detrimental to hVDAC-2 structure. Unfolding and aggregation process are sequential events and show strong temperature dependence. We demonstrate that an optimal lipid-to-protein ratio of 2600∶1 – 13000∶1 offers the highest protection against thermal denaturation. Activation energies derived only for lower LPRs are ∼17 kcal mol⁻¹ for full-length hVDAC-2 and ∼23 kcal mol⁻¹ for the Cys-less mutant, suggesting that the nine cysteine residues of hVDAC-2 impart additional malleability to the barrel scaffold. Our studies reveal that cysteine residues play a key role in the kinetic stability of the protein, determine barrel rigidity and thereby give rise to strong micellar association of hVDAC-2. Non-linearity of the Arrhenius plot at high LPRs coupled with observation of protein aggregation upon thermal denaturation indicates that contributions from both kinetic and thermodynamic components stabilize the 19-stranded β-barrel. Lipid-protein interaction and the linked kinetic contribution to free energy of the folded protein are together expected to play a key role in hVDAC-2 recycling and the functional switch at the onset of apoptosis.