Stability and motion of a hairpin and the corresponding mismatched duplex: a theoretical exploration using molecular mechanics and normal mode analysis of 2D NMR results on d(GCCGCAGC)

The oligomer d(GCCGCAGC) can adopt two different conformations: i) a duplex with two mismatched A.C base pairs and ii) a hairpin with two C.G base pairs and a single stranded loop. We report molecular mechanics, normal mode analysis, and thermodynamic stability calculations for both structures. We show that the energy-minimized structure and harmonic-dynamics results are in complete agreement with the observed NOE spectrum and imino proton exchange data. We conclude that the high stability of the hairpin structure over the duplex at low salt concentration is due to the higher vibrational entropy contribution to the system free energy by the single stranded loop and to the lack of minor groove phosphate/phosphate electrostatic repulsions that tend to destabilize the duplex.     

Conformational microheterogeneity in a DNA double helix: structure of restriction endonuclease Bam H1 recognition site

Structural studies using 500 MHz 1H NMR spectroscopy on Bam H1 recognition site d(GGATCC)2 in solution at 19 degrees is reported. The resonances from the sugar ring and base protons have been assigned from the 2D-COSY and NOESY spectra. Analyses of the NOESY cross-peaks between the base protons H8/H6 and sugar protons H2'/H2", H3' reveal that the nucleotide units G2, A3 and C6 adopt (C3'-endo, chi = 200 degrees-220 degrees) conformation while G1, T4 and C5 exhibit (C2'-endo, chi = 240 degrees-260 degrees) conformation. NMR data clearly suggest that the two strands of d(GGATCC)2 are conformationally equivalent and there is a structural two-fold between the two A-T pairs. The above information and the NOESY data are used to generate a structural model of d(GGATCC)2. The important features are: (i) G1-G2 stack, the site of cleavage, shows an alternation in sugar pucker i.e. C2'-endo, C3'-endo as in a B-A junction, (ii) G2-A3 stack adopts a mini A-DNA, both the sugars being C3'-endo, (iii) A3-T4 stack, the site of two-fold, displays an A-B junction with alternation in sugar pucker as C3'-endo, C2'-endo, (iv) T4-C5 stack adopts a mini B-DNA both the sugars being C2'-endo and (v) C5-C6 stack exhibits a B-A junction with C2'-endo, C3'-endo sugar puckers. Thus, our studies demonstrate that conformational microheterogeneity with a structural two fold, is present in the Bam H1 recognition site.     

Phenolic acid effects on peanut growth and oil production

Plants of peanut (Arachis hypogaea L. var. PG No. 1) were given two foliar sprays of phenolic compounds (H-acid, 1, 2, 4-acid, resorcinol and RD-Brown) at 100 and 200 ppm, 35 and 50 days after sowing. In treated plants, shelling %, yield (kg/ha), number of gynophores per plant and number of pods per plant were significantly greater than in the control. Oil content of kernels also showed a significant increase with all the phenolic compounds applied. These compounds increased the linoleic acid concentration so improving nutritional quality. The number of gynophores was significantly correlated with the number of pods per plant and yield per hectare. The effect of phenolic compounds on growth and development was independent of their structural configuration.     

MECHANISM OF ACTION OF β-N-OXALYL-L-α, β-DIAMINOPROPIONIC ACID, THE LATHYRUS SATIVUS NEUROTOXIN

The source of ammonia in the brain tissue of young rats treated with β-N-oxalyl-l -α, β-diaminopropionic acid (ODAP) has been studied. ODAP administration to 12-day-old rats causes a significant increase in the levels of adenylic acid deaminase in the brain. Glutaminase activity also shows an increase under these conditions. An increase in the levels of acid protease and transglutaminase is also observed in the brain of ODAP-treated animals. Glutamate dehydrogenase activity is decreased slightly. Glutamine synthetase enzyme is not affected. Aspartate-α-ketoglutarate transaminase and aspartate-pyruvate transaminase activities are enhanced in the brain tissue of ODAP-treated rats. It is held that protein degradation, especially the cleavage of free and protein-bound amide bonds, may be responsible for excess ammonia liberation in the brain of ODAP-treated young rats.     

Nuclear magnetic resonance and thermal studies of drug doped dipalmitoyl phosphatidyl choline-H2O systems

The influence of the sulfone drugs, diamino diphenyl sulfone and diamino monophenyl sulfone on the phase transitions and dynamics of dipalmitoyl phosphatidyl choline-H₂O/D₂O vesicles have been investigated using differential scanning calorimetry and nuclear magnetic resonance. Our results show that diamino diphenyl sulfone interacts quite strongly with the headgroups of dipalmitoyl phosphatidyl choline whereas the diamino monophenyl sulfone-dipalmitoyl phosphatidyl choline interaction is quite weak. This is attributed to the difference in the structure and hydrophobic character of the two drugs.     

A simple haemolytic method for quantitation of the delta endotoxin of Bacillus thuringiensis subsp. israelensis from crude samples

A simple haemolytic assay method for quantitative estimation of the delta endotoxin of Bacillus thuringiensis subsp. israelensis from a crude preparation has been developed. The method has several advantages over mosquito-larvicidal methods of assay as it is inexpensive, highly sensitive and easier to run and can be used for performing a reasonably large number of assays rapidly with high precision and with a coefficient of variation that does not exceed 1.96%.     

Mitogenic response of rat lung and tracheal epithelial cells in monolayer primary cultures—Modulation of TNF-α expression

Epithelial cells isolated from rat lung and trachea were grown on monolayers and their response to a number of hormones and growth factors were studied. Maximum proliferative response in serum containing media was observed when insulin, cholera toxin and cortisol were present together. However, these additives when present independently showed a marginal response. The synergism, due to these factors in promoting growth was seen very early in culture (day 4) as shown by thymidine labelling studies, On examining the indices of early mitogenesis, such as the expression ofc-myc, our data suggests that these factors stimulate the expression ofc-myc within 4 h. With respect to expression of TNF-α mRNA, this study suggests a possible modulation of TNF-α expression in response to these mitogens that stimulate proliferation maximally. Whether this expression of TNF-α by these epithelial cells is due to a maximal proliferative stimulus and/or is an early step in the cascade of intracellular signalling events is to be investigated in detail.     

Isolation and characterization of temperature-sensitive mutants ofAnabaena variabilis impaired in nitrogen fixation

Temperature-sensitive (ts) mutants of the cyanobacteriumAnabaena variabilis ATCC 29413 were isolated following mutagenesis with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and post-treatment with metronidazole at 40°C. Of the 8000 clones isolated and tested, six mutants were conditionally lethal at the restrictive temperature (40°C). All the ts mutants exhibited differences in their rates of growth, chlorophyll content, pigment (phycocyanin and/or chlorophyll) ratios, heterocyst frequency, oxygen evolution and nitrogenase activity at the permissive temperature (28°C). A gradual loss of all the above features occurred after a period of 3 d at 40°C, followed by lysis of the cultures. Cessation of nitrogenase activity was found to be different in the different ts mutants. The temperature-sensitive nature of the mutants is suggested to be due to an impairment in iron metabolism since addition of ferric citrate to cultures at 40°C restored the ability to grow, produce heterocysts and fix nitrogen.     

Photoheterotrophic and chemoheterotrophic dinitrogen fixation and nitrate utilization by the cyanobacteriumAnabaena torulosa

Anabaena torulosa exhibited fructose-dependent growth, heterocyst differentiation and N₂ fixation in nitrate-free (diazotrophic) cultures in photoheterotrophic and chemoheterotrophic conditions. The incorporation of nitrate into such cultures inhibited the formation of heterocysts and N₂ fixation. The rate of NO ₃ ⁻ uptake byA. torulosa in photoautotrophic, photoheterotrophic and chemoheterotrophic conditions was similar but it increased by 100% in phototrophic conditions. The activity of glucose-6-phosphate dehydrogenase was found to be maximum in phototrophic and photoheterotrophic conditions. Ferredoxin-NADP⁺ reductase, nitrate reductase and glutamate-ammonia ligase activities suggest that nitrate utilization takes place in nonphotosynthetic conditions.