N-Acetyl-D-prolinol

Francisella noatunensis subsp. orientalis (Fno) is the causative agent of piscine francisellosis, an emerging infectious disease in Asia and Latin America. In this study two outbreaks of francisellosis were diagnosed in the UK on the basis of histopathology, electron microscopy, PCR, bacterial isolation and fulfillment of Koch's postulates. Furthermore, a phenotypic fingerprint based on biochemical analyses, metabolic activity, chemotaxonomic composition, and antimicrobial assays was generated for the novel isolates, the Fno type strain Ehime-1 from Asia and other Fno from Latin America. The genetic relatedness between the novel Fno and other Francisellaceae species was investigated by sequencing and comparing the 16SrRNA gene, 8 housekeeping genes (individually and concatenated) and the 16SrRNA-ITS-23SrRNA sequence. The phenotypic profiling indicated a high degree of similarity among the Fno strains as all were able to metabolize dextrin, N-acetyl-D glucosamine, D-fructose, α-D-glucose, D-mannose, methyl pyruvate, acetic acid, α-keto butyric acid, L-alaninamide, L-alanine, L-alanylglycine, L-asparagine, L-glutamic acid, L-proline, L-serine, L-threonine, inosine, uridine, glycerol, D L-α-glycerol phosphate, glucose-1-phosphate, and glucose-6-phosphate. The chemotaxonomic analyses indicated that 24:1 (20.3%), 18:1n-9 (16.9%), 24:0 (13.1%) 14:0 (10.9%), 22:0 (7.8%), 16:0 (7.6%), and 18:0 (5.5%) were the predominant structural fatty acids in Fno. The antimicrobial assays showed little variation between the isolates and high susceptibility to enrofloxacin, gentamicin, neomycin, streptomycin, amikacin, ciprofloxacin, gatifloxacin, nitrofurantoin, tobramycin, kanamycin, tetracycline, oxytetracycline, florfenicol, oxolinic acid, and streptomycin in all the Fno analyzed. In all the phylogenetic trees the Fno strains clustered together in independent branches confirming a high degree of homogeneity. Interestingly in five of the 11 trees i.e., mutS, putA, rpoB, 16SrRNA-ITS-23SrRNA, and concatenated sequence the two Francisella noatunensis ssp. diverged more from each other than from the closely related Francisella philomiragia (Fp). The phenotypic and genetic characterization confirmed the Fno isolates represent a solid phylo-phenetic taxon that in the current context of the genus seems to be misplaced within the species Fn. We propose the use of the present polyphasic approach in future studies to characterize strains of Fnn and Fp and verify their current taxonomic rank of Fno and other aquatic Francisella spp.

Salinity hinders agricultural productivity worldwide by distressing plant metabolism. Growth of blackgram (Vigna mungo L. var. Sulata), an adverse climate-resistant pulse, is arrested under salinity. Present research integrates study of physio-biochemical parameters and non-targeted metabolomics approach to explore the alterations in metabolic pathway during adaptive responses of nonprimed and haloprimed blackgram seedlings grown hydroponically under NaCl stress. Salinity provoked accumulation of peroxides, compatible solutes and phenolics which increased free radical scavenging activities of nonprimed seedlings under salinity. Pre-germination seed halopriming abrogated NaCl-mediated adversities in haloprimed plantlets favouring better growth. Thus, farmers may adopt seed halopriming technique to improve blackgram productivity in saline-prone fields. Additionally, metabolomics study uncovered numerous metabolites amongst which 35 compounds altered significantly under salinity. The candidate metabolites were aspartic acid, L-glutamic acid, L-proline, L-asparagine, DL-isoleucine, L-homoserine, citrulline, L-ornithine, D-altrose, D-allose, N-acetyl-D-mannosamine, fructose, tagatose, sucrose, D-glucose, maltose, glycerol-1-phosphate, D-sorbitol, benzoic acid, shikimic acid, 4-hydroxycinnamic acid, arbutin, succinic acid, pipecolic acid, fumaric acid, nicotinic acid, L-pyroglutamic acid, oxalic acid, glyceric acid, maleamic acid, adenine, guanosine, lauric acid, stearic acid and porphine. Comparing metabolic responses of nonprimed and haloprimed seedlings, it was clear that efficient alteration in carbohydrate metabolism, phenolics accumulation, amino acid, organic acid and nucleic acid metabolism were the key places of metabolic reprogramming for tolerating salinity. Overall, we report, for the first time, 35 contributory candidate compounds that constituted core fundamental metabolome invoking salinity tolerance in nonprimed and haloprimed blackgram. These metabolites may be targeted by biotechnologists to produce high vigour salt-tolerant transgenic blackgram via genetic engineering.

In this study, we showed that the dual addition of glucosyl ceramide synthase and ceramidase inhibitors to A549 cell culture led to the possibility of ceramide channel formation via endogenous palmitoyl-ceramide accumulation with an increase in cholesterol contents in the lysosome membrane as an initial step prior to initiation of necrotic cell death. In addition, the dual addition led to black circular structures of 10-20 nm, interpreted as stain-filled cylindrical channels on transmission electron microscopy. The formation of palmitoyl-ceramide channels in the lysosome membrane causes the liberation of cathepsin B from lysosomes for necrotic cell death. On the other hand, necrotic cell death in the dual addition was not caused by oxidative stress or cathepsin B activity, and the cell death was free from the contribution of the translation of Bax protein to the lysosome membrane.