Interaction of curcumin with phosphatidylcholine: A spectrofluorometric study

Curcumin [1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3, 5-dione], the main constituent of the rhizomes of the plant Curcume longa L. (turmeric), is a powerful antioxidant in both enzymatic and nonenzymatic systems. The interactions of curcumin with egg and soy phosphatidylcholine were followed by fluorescence spectroscopy. Curcumin had very weak fluorescence in aqueous system, which was enhanced in apolar environments. Curcumin emitted at 490 nm after being excited at 451 nm in phosphatidylcholine micelles. The equilibrium constants for the interaction of curcumin with egg and soy phosphatidylcholine were (3.26 +/- 0.2) x 10(5) and (2.64 +/- 0.2) x 10(5) M(-1), respectively. From the Scatchard plot of the fluorometric data, it was inferred that one molecule of curcumin could bind six molecules of phosphatidylcholine. The equilibrium constant for the phosphatidylcholine-curcumin interaction decreased with temperature, indicating the amphiphilic nature of curcumin. The DeltaG, DeltaH, and DeltaS values obtained for the interaction of egg phosphatidylcholine-curcumin were -7.8 +/- 0.3 kcal/mol, -9.6 +/- 0.4 kcal/mol, and -6.8 +/- 0.2 cal/mol/K, respectively. The fluorescence anisotropy measurements of curcumin with phosphatidylcholine suggested that the anisotropy of the curcumin molecule did not change in phosphatidylcholine. The interaction of divalent metal ions with phosphatidylcholine-curcumin in comparison with phosphatidylcholine-1-anilino-8-naphathalenesulfonic acid complex suggested the strong binding of curcumin to metal ions.     

Rhizobacteria‐mediated resistance against the blackeye cowpea mosaic strain of bean common mosaic virus in cowpea (Vigna unguiculata)

BACKGROUND: The present study investigated the effect of seven Bacillus‐species plant‐growth‐promoting rhizobacteria (PGPR) seed treatments on the induction of disease resistance in cowpea against mosaic disease caused by the blackeye cowpea mosaic strain of bean common mosaic virus (BCMV). RESULTS: Initially, although all PGPR strains recorded significant enhancement of seed germination and seedling vigour, GBO3 and T4 strains were very promising. In general, all strains gave reduced BCMV incidence compared with the non‐bacterised control, both under screen‐house and under field conditions. Cowpea seeds treated with Bacillus pumilus (T4) and Bacillus subtilis (GBO3) strains offered protection of 42 and 41% against BCMV under screen‐house conditions. Under field conditions, strain GBO3 offered 34% protection against BCMV. The protection offered by PGPR strains against BCMV was evaluated by indirect enzyme‐linked immunosorbent assay (ELISA), with lowest immunoreactive values recorded in cowpea seeds treated with strains GBO3 and T4 in comparison with the non‐bacterised control. In addition, it was observed that strain combination worked better in inducing resistance than individual strains. Cowpea seeds treated with a combination of strains GBO3 + T4 registered the highest protection against BCMV. CONCLUSION: PGPR strains were effective in protecting cowpea plants against BCMV under both screen‐house and field conditions by inducing resistance against the virus. Thus, it is proposed that PGPR strains, particularly GBO3, could be potential inducers against BCMV and growth enhancers in cowpea. Copyright © 2009 Society of Chemical Industry     

High-level expression of SAG1 and GRA7 gene of Toxoplasma gondii (Izatnagar isolate) and their application in serodiagnosis of goat toxoplasmosis

Toxoplasmosis, caused by Toxoplasma gondii, is a disease of economic importance in livestock, especially in sheep and goats, where it causes abortion. Although several serological tests are in use for diagnosis of infection, production of reliable reagents is a constraint. An 814 bp sequence coding for a truncated surface antigen surface antigen 1 (SAG1), a tachyzoite stage-specific protein, as well as a 657 bp sequence coding for granule protein 7 (GRA7), a dense granule protein were PCR amplified from the genomic DNA of T. gondii. The amplified products were ligated in pET-32b(+) and pET-32c(+) expression vectors, respectively and subsequently transformed into BL21(DE3)pLysS cells. A high-level expression of the histidine-tagged SAG1 and GRA7 fusion proteins were obtained after 7h of incubation. The recombinant proteins were purified using Ni-NTA column and were characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis using reference positive sera from goat, rabbit and humans at 1:100 dilution. Subsequently, the diagnostic efficiency of the recombinant proteins, either individually or as a cocktail of the recombinant proteins, was assessed with 56 reference goat sera by enzyme-linked immunosorbent assay (ELISA). The immunoreactivity of the refolded SAG1 and GRA7 was evidenced by high OD values. The reactivity of the recombinant proteins as a cocktail preparation was more than that of individual proteins in ELISA and could detect accurately the infection in goats. This is the first report of serological detection of caprine toxoplasmosis by ELISA using a cocktail of recombinant Toxoplasma proteins.     

Local versus expert knowledge in forest management in a semi‐arid part of India

This study analyzes the differences between the villagers' and the Forest Department's preferences in the choice of tree species to be planted in eastern Rajasthan, India. Locals learned to manage the tree resources in a sustainable manner before the formal knowledge perspectives started to dominate through various state forestry programmes in which decision making is dominated by expert perspectives in a given political context. The study also attempts to identify the problems of complementary relationships between local and expert knowledge and the resulting management implications. Copyright © 2004 John Wiley & Sons, Ltd.     

Harmonised monitoring of Lake Macro Prespa as a basis for Integrated Water Resources Management

In this paper, reliable data and information as an important basis for integrated water resources management (IWRM) is discussed. Such data are especially important in transboundary water bodies, as they can represent a common and transparent knowledge-base that all concerned parties can trust and utilise in the management process. The transboundary Lake Macro Prespa (West Balkan) is used as a case study. The lake is rich in biodiversity but threatened by increased eutrophication. The main focus of the monitoring (2007–2008) was therefore on nutrients and chlorophyll a, as well as oxygen levels, transparency and temperature. The results of the monitoring were used in consultations with water managers and other stakeholders, and the study demonstrated how such data could represent a common and neutral point of departure for discussions across borders on integrated lake management. However, the paper also shows that monitoring may result in inadequate or even erroneous information, thus pointing to the importance of understanding all phases of the practical implementation of harmonised monitoring. The main challenges of the harmonised monitoring included different laboratory methods and performances, as well as a number of practical issues related to logistics and communications across borders. A less ambitious programme during the first year of monitoring (2007) might have saved funds and resulted in more harmonised and reliable data this year. However, in 2008 the project succeeded in producing a set of harmonised data, amongst others by using one common and accredited laboratory for stations in both countries. This paper therefore also presents new monitoring data from this unique large lake, of which there are relatively few published studies. The results of the monitoring give reasons for concern. The eutrophication of the lake seems to be increasing, and this calls for improved integrated management in order to reduce nutrient levels and improve the environmental status.     

Syntheses and application of anionic polyelectrolytes in water and waste water treatment

Five anionic polyelectrolytes were prepared with different molar ratios of acrylamide and acrylic acid monomers. The optimum flocculation conditions were found to depend on the polyelectrolyte and characteristics of the sample. The optimum time for flash mixing for alum and for polyelectrolytes was 1 to 2 min. The optimum flocculation time was 10 to 25 min and optimum flocculation intensity varied between 20 to 30 rpm. The pH was found to be inversely proportional to flocculation efficiency. The COD removal and coliform bacteria removal were 65 and 98%, respectively.     

Physical behavior of purified and crude wax obtained from Sunflower (Helianthus annuus) seed oil refineries and seed hulls

The sunflower seed waxes obtained from two sources (i) seed hull as astandard and (ii) crude wax from oil refineries were studied for theircrystallization, melting characteristics and morphology of crystals. Theresults of differential scanning calorimetry of wax obtained from seed hullsshowed the melting temperature range of 13.18 ^°C with the onsetat 62.32 ^°C, for purified wax, compared to the melting range of24.73 ^°C with the onset at 42.3 ^°C, for crude wax. Theenthalpy of fusion for both waxes were 57.55 mcal/mg and 7.63 mcal/mg,respectively. The DSC melt crystallization temperature range was15.79 ^°C with the onset of 64.58 ^°C for purified wax andtemperature range of 31.45 ^°C with an onset of 57.76 ^°C forcrude wax. A similar pattern was observed of wax obtained from thecrude wax of oil refineries. The enthalpy of crystallization was –64.27mcal/mg and –7.67 mcal/mg, respectively. The purified wax obtained fromthe two sources (i) and (ii) were comparable with completion temperaturesof 75.5 ^°C and 75.1 ^°C, respectively. The effect of inhibitor(lecithin) on crystallization of purified wax under light microscope andsurface structure by scanning electron microscope were observed. Lecithinat 0.2% inhibited the crystallization but nucleation was unaltered. Thewax crystal was inhibited to around 60% of the original size with 0.2%lecithin. It is concluded that the sunflower waxes studied were notcomparable in their crystal properties of crude and purified states.Lecithin inhibited the crystallization of sunflower seed wax.     

Delineation of Inheritance Pattern of Aleurone Layer Colour Through Chemical Tests in Rice

Background

Rice aleurone layer develops different colours with various chemical tests that may help to develop some rapid tests for identification/grouping of rice varieties. Understanding the colour inheritance pattern could enable to develop chemical clues that may help for genetic purity analysis along with grow-out-test.

Results

In this study, inheritance pattern of aleurone layer colour was studied in parents, F₁ and F₂ progenies derived from the crosses IR 36 × Acc. No. 2693 and IR 64 × Acc. No. 2693. The parent IR 36 showed light yellow (NaOH/KOH) and brown (phenol/modified phenol test) colour; whereas, Acc. No. 2693 revealed wine red/dark wine red (NaOH/KOH) and light brown colour/no reaction (phenol/modified phenol test). In contrary, another parent IR 64 exhibited light yellow (KOH/NaOH) and dark brown (phenol, modified phenol) colour. Both the F₁ showed an intermediate light wine red colour (NaOH/KOH) and dark brown (phenol and modified phenol) colour, which is dominant over their one of the parents. The colour pattern with standard phenol/modified phenol, NaOH and KOH tests in F₂ progenies of both the crosses showed 9:7 (complementary gene interaction) and 11:5 ratios (reciprocal dominance modification of recessive alleles), respectively.

Conclusions

Our findings clearly elucidate the colour inheritance pattern in rice that may facilitate to develop rapid chemical tests to identify/ group the varieties for genetic purity analysis.

     

Efficacy of anti-staphylococcal protein P128 for the treatment of canine pyoderma: potential applications

In this study, we demonstrate the antibacterial activity of P128 on Staphylococcus isolates responsible for canine pyoderma. Eighty seven swabs were collected from dogs suffering from pyoderma and subjected to antibiotic sensitivity test and 46 Staphylococcus strains were isolated and characterized. In-vitro antimicrobial susceptibility testing with P128 was done by Minimum Inhibitory Concentration (MIC) method as per CLSI guidelines. All the Staphylococci isolated from the dogs with pyoderma, although showed resistance to various antibiotics tested, were lysed by P128. Clinical efficacy of P128 was examined in 17 dogs with pyoderma by application of the P128 hydrogel twice daily for 8 days and the results indicated complete healing of all the lesions of all the dogs under treatment. Under the conditions of this study, P128 was found to be a potent convenient proteinaceous drug for the treatment of staphylococcal pyoderma in dogs.