Occurrence of Horizontal Gene Transfer of PIB-type ATPase Genes among Bacteria Isolated from the Uranium Rich Deposit of Domiasiat in North East India

Uranium (U) tolerant aerobic heterotrophs were isolated from the subsurface soils of one of the pre-mined U-rich deposits at Domiasiat located in the north-eastern part of India. On screening of genomic DNA from 62 isolates exhibiting superior U and heavy metal tolerance, 32 isolates were found to be positive for P_IB-type ATPase genes. Phylogenetic incongruence and anomalous DNA base compositions revealed the acquisition of P_IB-type ATPase genes by six isolates through horizontal gene transfer (HGT). Three of these instances of HGT appeared to have occurred at inter-phylum level and the other three instances indicated to have taken place at intra-phylum level. This study provides an insight into one of the possible survival strategies that bacteria might employ to adapt to environments rich in uranium and heavy metals.     

Changes in Membrane Fluidity and Protein Composition during Release of Cucumber Seeds from Dormancy by a Higher Temperature Shift

Dormancy of seeds of cucumber (Cucumis sativus L.) was inducedby imbibing in -1.8 MPa polyethylene glycol 6000 (PEG) solutionand pulsing with far red light for 15 min prior to washing anddrying. When re-imbibed with water at 20 °C, dormancy wasbroken by raising the temperature to 30 °C for 6 h. Thistreatment was also effective when -0.9 MPa PEG was present duringre-imbibition and high temperature. Seeds with broken dormancywere found to germinate in water over a smaller temperaturerange than seeds in which dormancy had not been induced. Whenthe duration of the temperature shift to 30 °C was varied,germination percentage increased from 7 to 60% after 6 h, butlonger exposures up to 12 h had no further promoting effect.The time course of germination after transfer to water following6 h at 30 °C in PEG showed piercing of the perisperm-endospermenvelope after 9–12 h and radicle protusion after 12–15h. If PEG was retained after high temperature treatment no visiblegermination was observed. Thus, to study membrane fluidity andthe protein content associated with germination, seeds weresampled 9 h after high temperature treatment. To study the germinablebut not germinating state, seed held in PEG for 9 h rather thanin water was used. Dormant seed was sampled before the hightemperature treatment. Membrane fluidity was assessed usingfluorescence polarization of membrane fractions treated withDPH (1,6-diphenyl-1,3,5-hexatriene) or its derivatives. Membraneproteins were compared using one-dimensional SDS-PAGE electrophoresis.Intracellular membrane fluidity was not increased in the transitionfrom the dormant to germinable state, but did increase in thetransition to germination. There were no detected changes inintracellular membrane proteins during either transition. Inplasma membrane fractions, fluidity increased during both transitions,while a marked increase in 21, 18 and 17 kD proteins was observedin the transition from germinable to germinating state. Thusmodification of plasma membrane fluidity rather than changesin protein profile is associated with the high temperature releaseof cucumber seeds from dormancy. Copyright 2000 Annals of BotanyCompany     

UDP-Gal: <Emphasis Type="Italic">N</Emphasis>-acetylglucosamine β 1–4 galactosyltransferase expressing live attenuated parasites as vaccine for visceral leishmaniasis

As compared to cutaneous leishmaniasis, vaccination against visceral leishmaniasis (VL) has received limited attention. In this study, we demonstrate for the first time that an UDP-Galactose: N-acetylglucosamine β 1–4 galactosyltransferase (GenBank Accession No. EF159943) expressing attenuated LD clonal population (A-LD) is able to confer protection against the experimental challenge with the virulent LD AG83 parasite. A-LD was also effective in established leishmania infection. The vaccinated animals showed both cell mediated (in vitro T-cell proliferation, and DTH response) and humoral responses (Th1 type). These results demonstrate the potential of the attenuated clones as an immunotherapeutic and immunoprophylactic agent against visceral leishmaniasis.     

Thermal and Structural Stability of Adsorbed Proteins

Experimental evidence suggests that proteins adsorbed to hydrophobic surfaces at low coverages are stabilized relative to the bulk. For larger coverages, proteins unfold and form β-sheets. We performed computer simulations on model proteins and found that: 1), For weakly adsorbing surfaces, unfolded conformations lose more entropy upon adsorption than folded ones. 2), The melting temperature, both in the bulk and at surfaces, decreases with increasing protein concentration because of favorable interprotein interactions. 3), Proteins in the bulk show large unfolding free energy barriers; this barrier decreases at stronger adsorbing surfaces. We conjecture that typical experimental temperatures appear to be below the bulk melting temperature for a single protein, but above the melting temperature for concentrated protein solutions. Purely thermodynamic factors then explain protein stabilization on adsorption at low concentrations. However, both thermodynamic and kinetic factors are important at higher concentrations. Thus, proteins in the bulk do not denature with increasing concentration due to large kinetic barriers, even though the aggregated state is thermodynamically preferred. However, they readily unfold upon adsorption, with the surface acting as a heterogeneous catalyst. The thermal behavior of proteins adsorbed to hydrophobic surfaces thus appears to follow behavior independent of their chemical specificity.     

A new species of Eriocaulon (Eriocaulaceae) from Madhya Pradesh, India

Eriocaulon raipurense (Eriocaulaceae) is described and illustrated as a new species from Madhya Pradesh, India. The species is closely allied to E. hamiltonianum but differs in the size and apex of involucral bancts, white-pilose nature of floral bracts and colour of female petals.