An Enzyme‐Linked Immunosorbent Assay (ELISA) for Detection of Marek's Disease Virus‐Specific Antibodies and its Application in an Experimental Vaccine Trial

An enzyme‐linked immunosorbent assay (ELISA) for the detection of Marek's disease virus (MDV)‐specific antibodies was developed. Chicken embryo cells (CEC) or chicken kidney cells (CKC) were infected with MDV vaccine strain CVI988/Rispens, and infected‐cell lysates were prepared at day 5 post‐infection by freeze‐thawing. Uninfected‐cell lysates served as negative controls. Sera were used at a 1 : 100 dilution and were added in parallel to wells containing the infected and uninfected cell lysates. The optical densities at 492 nm (OD_{492 nm}) were measured after detection of bound chicken antibodies with anti‐chicken IgG peroxidase conjugate and colour reactions using o‐phenylenediamine (OPD) as a substrate. The best results concerning the signal‐to‐noise ratio were obtained by using CKC cells rather than CEC for antigen preparation. The OD_{492 nm} of plasma or serum samples with infected CKC was <0.02 when samples of unvaccinated and unchallenged maternal antibody‐negative white leghorn chickens were tested. Sera and plasma samples of positive control birds exhibited OD_{492 nm} of <0.01 when tested with uninfected CKC. The assay was used to monitor a trial that compared experimental BAC DNA vaccines and a commercial vaccine. Sustained seroconversion and antibody titers that were constantly rising until day 84 after vaccination (71 days after challenge) was observed only when chickens did not develop Marek's disease. In contrast, chickens developing the disease mounted marginal and short‐lived antibody titers only. We conclude that the developed ELISA may be a valuable tool for the evaluation of the efficacy of MDV vaccination under experimental but possibly also under field conditions.     

Biologically active carbazole alkaloids from Murraya koenigii

The bioassay guided fractionation of the acetone extract of the fresh leaves of Murraya koenigii resulted in the isolation of three bioactive carbazole alkaloids, mahanimbine (1), murrayanol (2), and mahanine (3), as confirmed from their (1)H and (13)C NMR spectral data. Compound 2 showed an IC(50) of 109 microg/mL against hPGHS-1 and an IC(50) of 218 microg/mL against hPGHS-2 in antiinflammatory assays, while compound 1 displayed antioxidant activity at 33.1 microg/mL. All three compounds were mosquitocidal and antimicrobial and exhibited topoisomerase I and II inhibition activities.     

Isolation and characterization of stelladerol, a new antioxidant naphthalene glycoside, and other antioxidant glycosides from edible daylily (hemerocallis) flowers.

Daylily (Hemerocallis spp.) flowers are utilized as an important ingredient in traditional Asian cuisine and are also valued for their reputed medicinal effects. In studies of the bioactive methanol and aqueous methanol extracts of lyophilized Hemerocallis cv. Stella de Oro flowers, kaempferol, quercetin, and isorhamnetin 3-O-glycosides (1-9), phenethyl beta-D-glucopyranoside (10), orcinol beta-D-glucopyranoside (11), phloretin 2'-O-beta-D-glucopyranoside (12), phloretin 2'-O-beta-D-xylopyranosyl-(1-->6)-beta-D-glucopyranoside (13), a new naphthalene glycoside, stelladerol (14), and an amino acid (longitubanine A) (15) have been isolated. All of these compounds were tested for their antioxidant and cyclooxygenase inhibitory activities. Stelladerol was found to possess strong antioxidant properties, inhibiting lipid oxidation by 94.6% +/- 1.4 at 10 microM in an in vitro assay. Several of the flavonol 3-O-glycoside isolates also demonstrated modest antioxidant activities at 10 microM. None of the isolates inhibited cyclooxygenase activity at 100 microM.     

Agro-biodiversity and ethnobotany of Lakshadweep Islands of India

The Lakshadweep is a tiny landmass of 32 km² area (8^o–12^oN, 71^o–74^oE) constituted by a group of 27 small atolls located in the Indian territory of Arabian Sea of Indian Ocean. Studies on crop genetic diversity including wild relatives of crop plants of the region are little known except for coconut palm. This report deals with 106 collections assembled in two exploration and collection missions comprising 46 plant species under 35 genera belonging to 24 families. The plant germplasm collected during the trips are mostly useful as vegetable, fibre, medicinal plant, dye, edible tuber, timber and ecosystem service provider. The crop wild relatives collected during the study include that of sunnhemp, melon, yam, jute, pigeon pea, sugarcane, etc. A set of germplasm accessions was sent to national research institutes for regeneration. Out of the total collection, 64 accessions were assigned with national identity (Indigenous Collection or IC numbers: IC 0572014 to IC 0572077), and conserved in long term storage at National Gene Bank or in the field gene bank of ICAR-NBPGR, New Delhi. The important germplasm assembled from the islands include highly pungent betel leaf and a large fruited noni. Plants of Cajanus scarabaeoides (L.) Thouars, bitter less Cucumis melo L. var. agrestis Naudin saline tolerant Canavalia cathartica Thouars, Vigna marina (Burm.) Merr. and Corchorus trilocularis L. were the unique wild relatives collected. The current report is the first of its kind on the exploration and collection of plant genetic resources (other than coconut) from Lakshadweep Islands. Wild melon accessions collected are currently employed in distant hybridization programs. During the survey, ethnobotanic information of 23 plants for medicinal and other uses was also documented. Novel uses of the native plant species in the islands indicate their potential for exploitation in other coastal ecosystems. This report aims to highlight the potential plant resources of the region for direct human use and in future breeding programs.     

Nitrification inhibitors from the roots of Leucaena leucocephala

The nitrification inhibition (NI) bioassay guided fractionation of the methanol extract of lyophilized and milled roots of Leuceana leucocephala resulted in the isolation of four compounds, 1-4, as confirmed from their 1H and 13C NMR spectral data. Compound 1, gallocatechin, was the most active NI inhibitor at 12 microg/mL. Epigallocatechin, 2, and epicatechin, 4, isolated as mixtures, were not assayed individually for their NI inhibitory activities against the nitrification bacterium Nitrosomonas europaea.     

Are women really more talkative than men?

Women are generally assumed to be more talkative than men. Data were analyzed from 396 participants who wore a voice recorder that sampled ambient sounds for several days. Participants' daily word use was extrapolated from the number of recorded words. Women and men both spoke about 16,000 words per day.     

Production and Characterization of Siderophores and its Application in Arsenic Removal from Contaminated Soil

Siderophores are small molecular weight extracellular organic compounds secreted by microorganisms under iron-starved conditions, used by them to chelate and solubilize iron. Though they are specific ferric iron chelator, but is reported that they bind other metals also, such as divalent heavy metals and actinides because of potentially high metal-siderophore stability constants. Thus metal contaminant fate and transport in subsurface environment can be heavily influenced by siderophores. This approach can be successfully used in removing many toxic metals off the soil which poses a serious health threat. Our research focuses on the correlation between cell growth and siderophore production and chemical characterization of the siderophore type. Its also documents the development of an assay method for the screening of different metals for complexation with siderophores based on the Chrome Azurol S (CAS) assay. The present research aims at batch scale mobilization of arsenic from arsenic contaminated soils using siderophore produced by P. azotoformans and thus evaluating its efficiency as compared to Ethylene Diamine Tetra Acetic Acid (EDTA), Citric Acid (CA) for the same. FT-IR spectroscopic studies were carried out to determine the interaction between soil, arsenic and siderophore. Results have shown that the cell growth and siderophore production are inversely related. Characterization of siderophore produced by P. azotoformans has revealed that it is of mixed-type catecholate and hydroxamate. Siderophore was found to complex with heavy metals like Cadmium, Lead, Nickel, Arsenic (III, V), Aluminium, Magnesium Zinc, Copper, Cobalt, Strontium other than Iron. Five washings by siderophore, EDTA, CA removed almost 92.8%, 77.3%, 70.0% arsenic respectively as compared to only 33.8% removal by control. Washing of arsenic contaminated soil with tap water revealed that ≈ 65.8% of arsenic in contaminated soil is in freely available or weakly bound form. The IR spectra revealed that hydrogen bonding exists between siderophore, arsenic and soil. Encouraging results of arsenic removal by biomolecule-siderophore can lead to an emerging tool brimming with opportunities for environmental clean up.     

An enzyme-linked immunosorbent assay (ELISA) for detection of Marek's disease virus-specific antibodies and its application in an experimental vaccine trial

An enzyme-linked immunosorbent assay (ELISA) for the detection of Marek's disease virus (MDV)-specific antibodies was developed. Chicken embryo cells (CEC) or chicken kidney cells (CKC) were infected with MDV vaccine strain CVI988/Rispens, and infected-cell lysates were prepared at day 5 post-infection by freeze-thawing. Uninfected-cell lysates served as negative controls. Sera were used at a 1 : 100 dilution and were added in parallel to wells containing the infected and uninfected cell lysates. The optical densities at 492 nm (OD(492 nm)) were measured after detection of bound chicken antibodies with anti-chicken IgG peroxidase conjugate and colour reactions using o-phenylenediamine (OPD) as a substrate. The best results concerning the signal-to-noise ratio were obtained by using CKC cells rather than CEC for antigen preparation. The OD(492 nm) of plasma or serum samples with infected CKC was <0.02 when samples of unvaccinated and unchallenged maternal antibody-negative white leghorn chickens were tested. Sera and plasma samples of positive control birds exhibited OD(492 nm) of <0.01 when tested with uninfected CKC. The assay was used to monitor a trial that compared experimental BAC DNA vaccines and a commercial vaccine. Sustained seroconversion and antibody titers that were constantly rising until day 84 after vaccination (71 days after challenge) was observed only when chickens did not develop Marek's disease. In contrast, chickens developing the disease mounted marginal and short-lived antibody titers only. We conclude that the developed ELISA may be a valuable tool for the evaluation of the efficacy of MDV vaccination under experimental but possibly also under field conditions.