Properties of an antibody to Kelevan isolated by affinity chromatography: Antibody reactivation of ATPase activities inhibited by pesticides

Antibody molecules were produced by injection of BSA-Kelevan into chickens and rabbits. Pure antibody was obtained by a single pass of blood serum through an affinity column. The affinity gel was prepared by covalently binding BGG-Kelevan to activated Sepharose 4B-CN. Purity of the antibody was determined by ultracentrifugation and gel electrophoresis. Properties of the antibody included: sedimentation coefficient = 6.2, pI = 7.0, calculated MW = 150,000, and precipitin band formation using the microouchterlony test. The antibodies in free or immobilized form were able to prevent or reverse Kepone inhibition of ATPase activity from a variety of tissues from different sources. About 70 μg (approx 0.4 μM) of purified antibody was sufficient to restore the activity of mitochondrial (oligomycin-sensitive) Mg²⁺ ATPase activity which had been inhibited (in vitro) by 1 μM Kepone. The antibody was effective in preventing enzyme inhibition by other organochlorine pesticides with widely differing molecular structures. However, nonchlorinated inhibitors of mitochondrial oligomycin-sensitive Mg²⁺ ATPase activity were much less affected by the antibody. The available evidence suggests that the antibody binding site for the hapten may be specific for secondary or induced bonding forces due to the carbon-chlorine bonds rather than for a specific molecular structure.     

Screening of Barley Varieties for Drought Resistance

Barley varieties were screened for their efficient drought enduring ability and associated characters which may be useful in breeding programmes. The characters studied were: relative water content, content of proline, chlorophyll stability index (CSI), and percentage grain loss per plant due to water stress subjected to 10 days from jointing to boot stage. Proline and chlorophyll stability index were greatly associated with drought response (r =+0.54; -0.49), flag leaf area (r =+0.69; -0.51) and, to a certain extent, with leaf number (r = -0.35 and +0.34). Among the studied 19 varieties of barley, only three (DL 192, DL 36, DL 3) showed high enduring ability, of these the former two fall in the early, and the last one in the late flowering group. Morphological architecture of DL 192 had gathering type tiller (narrow tiller angle), wider and floppy leaves and it also bears physiological characters (high proline and high relative water content, and low value of chlorophyll stability index) necessary for drought resistance. DL 3 and DL 36 had dispersing type tiller (wider tiller angle) and erect leaf with good leaf area/plant, but unit leaf area was low in DL 3 (relates with shading behaviour reducing water loss). Other relations of drought resistance are briefly discussed in the text. An appraisal of the various traits revealed that high proline content, and low CSI (biochemical characters), earliness, larger flag leaf area and fewer leaves per plant (morphological characters) were directly correlated with productivity of the cultivars under drought conditions. These characters could be used as a guideline for breeding drought resistant cultivars of barley.     

Dissipation and distribution behavior of azoxystrobin, carbendazim, and difenoconazole in pomegranate fruits

The dissipation behavior and degradation kinetics of azoxystrobin, carbendazim, and difenoconazole in pomegranate are reported. Twenty fruits/hectare (5 kg) were collected at random, ensuring sample-to-sample relative standard deviation (RSD) within 20-25%. Each fruit was cut into eight equal portions, and two diagonal pieces per fruit were drawn and combined to constitute the laboratory sample, resulting in RSDs <6% (n = 6). Crushed sample (15 g) was extracted with 10 mL of ethyl acetate (+ 10 g Na(2)SO(4)), cleaned by dispersive solid phase extraction on primary secondary amine (25 mg) and C(18) (25 mg), and measured by liquid chromatography tandem mass spectrometry. The limit of quantification was ≤0.0025 μg g(-1) for all the three fungicides, with calibration linearity in the concentration range of 0.001-0.025 μg mL(-1) (r(2) ≥ 0.999). The recoveries of each chemical were 75-110% at 0.0025, 0.005, and 0.010 μg g(-1) with intralaboratory Horwitz ratio <0.32 at 0.0025 μg g(-1). Variable matrix effects were recorded in different fruit parts viz rind, albedo, membrane, and arils, which could be correlated to their biochemical constituents as evidenced from accurate mass measurements on a Q-ToF LC-MS. The residues of carbendazim and difenoconazole were confined within the outer rind of pomegranate; however, azoxystrobin penetrated into the inner fruit parts. The dissipation of azoxystrobin, carbendazim, and difenoconazole followed first + first order kinetics at both standard and double doses, with preharvest intervals being 9, 60, and 26 days at standard dose. At double dose, the preharvest intervals extended to 20.5, 100, and 60 days, respectively.     

Processing of commercial peanut cake into food-grade meal and its utilization in preparation of cookies

The commercial cake produced during expeller pressing of peanuts was extracted with n-hexane, and 80% ethanol followed by sieving through 80 mesh, to remove residual oil, pigments, bitter taste and fibrous material. The processed meal exhibited comparable composition with defatted peanut flour prepared in the laboratory by solvent extraction of full-fat peanut meal. However, the processed cake meal exhibited low methionine content and in vitro protein digestibility as compared with defatted peanut flour. The processed cake meal can be blended with wheat flour to the extent of 10% (w/w) to prepare acceptable cookies with improved protein and mineral contents.     

Cloning and phylogenetic analysis of Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) from Indian dromedaries (Camelus dromedarius)

The cDNAs of two proinflammatory cytokines viz., IL-6 and TNF-α from dromedarian camels were amplified by PCR using bactrian camel sequences and subsequently cloned for sequence analysis. Relationship based on amino acid revealed that dromedarian camel IL-6 shared 99.5% identity both at nucleotide and amino acid level with bactrian camel IL-6 and in case of TNF-α, the identity of dromedarian camel was 99.4% and 99.1% at nucleotide and amino acid level, respectively with that of bactrian camel. Phylogenetic analysis based on their amino acid sequences indicated the close relationship in these cytokine genes between dromedarian camel and other members of camelids.     

A major QTL for plant height is linked with bloom locus in sorghum [Sorghum bicolor (L.) Moench]

Plant height is one of the most important agronomic traits in sorghum with its relevance for biomass, grain yield, fodder and lodging. To understand its genetic basis, the quantitative trait loci (QTL) were identified using a recombinant inbred line (RIL) population consisting of 168 RILs derived from the cross between the two sorghum inbred lines 296B (dwarf) and IS18551 (tall) over six seasons. Two major QTL were identified one each on chromosomes SBI-06 and SBI-07 corresponding to the Dw2 and Dw3 gene loci together accounting 41 % plant height variation. In addition, a morphological bloom trait locus which remained unlinked in the linkage map was found to be significantly linked with plant height in single marker analysis explaining 22 % of the trait variation. By comparing the map positions of Dw1, Dw2 and Dw3, the new locus for plant height linked with bloom is proposed as Dw4 locus. Both SSR and the morphological bloom loci linked with height QTL of the present study can be employed as effective tools in marker-assisted breeding for rapid conversion of selected inbred parent lines either as dwarf seed (male sterile) parents or taller pollinators for hybrid seed industry, or for developing high biomass lines in sweet sorghum for exploitation as high bio-fuel crop.