Immunogenic and antigenic properties of recombinant soluble glycoprotein of rabies virus

Rabies virus glycoprotein is a type I transmembrane protein exposed on the surface on the mature virus particle that induces virus neutralizing antibodies. In the present study, 60 amino acid C-terminal hydrophobic anchor (transmembrane) and cytoplasmic domains of glycoprotein were deleted from full-length glycoprotein and fused with polyhistidine tag. The N-terminal viral signal peptide was also replaced with CD33 signal peptide for efficient secretion in mammalian cells. Following transfection of Madin Darby bovine kidney (MDBK) cells with plasmid encoding this soluble form of glycoprotein, polyclonal populations of stably transfected resistant cells were obtained after G418 selection. The protein was expressed as a glycosylated protein and secreted outside the cells utilizing N-terminal CD33 signal peptide. The secreted soluble glycoprotein was purified from cell culture supernatant by Ni--agarose affinity chromatography utilizing C-terminal polyhistidine tag. Like full-length glycoprotein, the expressed recombinant soluble glycoprotein was found to be immunogenic when injected in rabbits. In this study, we have assessed the potential of recombinant soluble glycoprotein as diagnostic antigen in ELISA and found that this recombinant protein can be used as diagnostic antigen in ELISA for detecting anti-glycoprotein antibodies in immunized host.     

Induction of <Emphasis Type="Italic">Fusarium</Emphasis> wilt (<Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">pisi</Emphasis>) resistance in garden pea using induced mutagenesis and in vitro selection techniques

Wilt caused by Fusarium oxysporum f. sp. pisi is a serious production constraint for peas worldwide. An attempt was made to isolate wilt-resistant mutants in two susceptible pea genotypes, Arkel and Azad P-1, employing induced mutagenesis and in vitro selection techniques. Two thousand seeds of each genotype were mutagenized either with ethyl methane sulfonate (EMS, 0.2% and 0.3%) or gamma rays (5-22.5 kR) in ⁶⁰Co gamma cell for three consecutive years. Screening of different mutagenized populations under wilt-sick plots resulted in the isolation of 25 mutants exhibiting complete or enhanced wilt resistance compared to parental genotypes. Five of these wilt-resistant mutants also outperformed the susceptible background genotypes in terms of yield and other horticultural traits. Efforts were also made to isolate wilt-resistant regenerants from callus cultures exhibiting insensitivity to culture filtrate (CF) of F. oxysporum f. sp. pisi. A total of 250 regenerants (R ₀) were obtained from CF-insensitive calli selected from medium supplemented with 20% culture filtrate. When evaluated in artificially inoculated sick plots, only five R ₂ lines obtained from the regenerants exhibited enhanced wilt resistance compared to parental cultivars. However, the selected lines did not exhibit resistance levels equivalent to those shown by wilt-resistant lines isolated through in vivo mutagenesis. To conclude, induced mutagenesis through irradiation and EMS treatments exhibited superiority over in vitro selection for inducing wilt resistance in peas.     

Resistance to bean common mosaic potyvirus strains and its inheritance in some Indian land races of common bean

Evaluation of 397 common bean accessions of diverse origin revealed the presence of effective sources of resistance against bean common mosaic potyvirus strains prevalent in Himachal Pradesh, a north-western Himalayan state of India. 21 accessions viz., KR 7, KR 225, KR 295, KRC 4, KRC 7, KRC 11, KRC 12, KRC 13, KRC 16, KRC 22, Amanda, Black Turtle Soup, Contender, Hans, Great Northern UI 123, Improved Tender Green 40031, Jubila, Kentucky wonder, Monroe, Premier and Sanilac, were found resistant to NL-1n and NL-7n strains. Some of the genotypes showed strain specific resistance. Pattern of inheritance studied in Hans, Contender, KRC 4 and KRC 22 of Indian origin against strain NL-1n revealed that single dominant gene governed the resistance in Hans and Contender whereas single recessive gene determined the resistance in KRC 22 and KRC 4. SCAR marker SW13₆₉₀ amplified a product of 690 bp in Hans and Contender, confirmed the presence of I gene in these cultivars. The implications of above genes in breeding for programme for BCMV resistance are discussed.     

Phenylpropanoids of Alpinia galanga as efflux pump inhibitors in Mycobacterium smegmatis mc(2) 155

The first and second line drugs used for the treatment of tuberculosis are now becoming ineffective due to emergence of resistant strains. Efflux pump provokes resistance in mycobacterium and hence could be explored as a new target for the discovery of anti-TB agents. In search of efflux pump inhibitors, MIC and modulation factor of phenylpropanoids isolated from A. galanga rhizome were determined prior to the accumulation and efflux assay. Phenylpropanoid compounds viz. 1'-S-1'-Acetoxychavicol acetate, trans-p-coumaryl diacetate and 1'-S-1'-acetoxyeugenol acetate were found to be potent modulators and decreased the MIC of ethidium bromide by 64 fold at the concentration of 2.5, 6.25 and 5.0mg/L respectively. 1'-S-1'-Acetoxyeugenol acetate enhanced the accumulation and inhibited the efflux of EtBr in Mycobacterium smegmatis mc(2) 155 cells.     

Detection of the occurrence of the fall armyworm,Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) strains on maize in Southern Rajasthan India

Journal of Plant Diseases and Protection - The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), a polyphagous lepidopteran pest that is native to tropical and sub-tropical America has...     

Verminous meningoencephalomyelitis in a red kangaroo associated with Angiostrongylus cantonensis infection

Angiostrongylus cantonensis is a zoonotic parasitic helminth that normally resides in the pulmonary arteries and the right ventricle of rats (Rattus sp.), the definitive host, where it causes little disease. Humans, dogs, opossums, and various zoo animals are “accidental” hosts. Here we report verminous meningoencephalomyelitis caused by A. cantonensis in a 9-mo-old male red kangaroo (Macropus rufus). The kangaroo was first presented lethargic, recumbent, and hypothermic, with severe muscle wasting. Within 3 wk, he progressed to non-ambulatory paraparesis and died. Gross examination revealed multifocal areas of dark-brown discoloration, malacia, and cavitation in the brain and the spinal cord. Histologically, there were several sections of nematodes surrounded by extensive areas of rarefaction, hemorrhage, spongiosis, neuronal necrosis, and gliosis. Based on size, morphology, and organ location, the nematodes were identified as subadult males and females. Interestingly, an eosinophilic response was largely absent, and the inflammatory response was minimal. A. cantonensis infection had not been reported previously in a red kangaroo in Louisiana or Mississippi, to our knowledge. Our case reaffirms the widespread presence of the helminth in the southeastern United States and indicates that A. cantonensis should be considered as a differential in macropods with neurologic clinical signs in regions where A. cantonensis is now endemic.     

Metabolomic Characterization of a cf. Neolyngbya Cyanobacterium from the South China Sea Reveals Wenchangamide A,a Lipopeptide with In Vitro Apoptotic Potential in Colon Cancer Cells

Metabolomics can be used to study complex mixtures of natural products, or secondary metabolites, for many different purposes. One productive application of metabolomics that has emerged in recent years is the guiding direction for isolating molecules with structural novelty through analysis of untargeted LC-MS/MS data. The metabolomics-driven investigation and bioassay-guided fractionation of a biomass assemblage from the South China Sea dominated by a marine filamentous cyanobacteria, cf. Neolyngbya sp., has led to the discovery of a natural product in this study, wenchangamide A (1). Wenchangamide A was found to concentration-dependently cause fast-onset apoptosis in HCT116 human colon cancer cells in vitro (24 h IC₅₀ = 38 μM). Untargeted metabolomics, by way of MS/MS molecular networking, was used further to generate a structural proposal for a new natural product analogue of 1, here coined wenchangamide B, which was present in the organic extract and bioactive sub-fractions of the biomass examined. The wenchangamides are of interest for anticancer drug discovery, and the characterization of these molecules will facilitate the future discovery of related natural products and development of synthetic analogues.     

Seed protein diversity in Solanum melongena L. and its wild and weedy relatives

Seed protein profiles by SDS-PAGE were studied in 72 accessions ofSolanum melongena L. and its related taxa in subgenusLeptostemonum (Dunal) Bitter. Comparisons based onJaccard's similarity and UPGMA clustering revealed interrelationships broadly inconformity with the conventional taxonomic treatments. Thus,S. viarum Dunal (sectionAcanthophora Dunal), S.torvum Swartz (section Torva Nees) andS. sisymbriifolium Lam. (sectionCryptocarpum Dunal) were located on separate branches ofthe dendrogram while all taxa of section Melongena Dunal,except S. macrocarpon Dunal, wereclustered closer together. S.melongena, incanum and insanum,all members of the eggplant complex, were revealed to bear very high similaritywith each other. Most of their accessions had identical band patterns supportingthe contention that these represent an interbreeding complex with limitedgenetic differentiation.