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Now a days the chemistry of Benzo[1,3]oxazine-2,4-dione moiety enhanced important due to their marvelous activities in medicinal field. Hence, some novel benzo[1,3]oxazine-2,4-dione derivatives were synthesized by various methods including eco-friendly methods like microwave irradiation and mortar pastel. These methods are then compared with conventional method. Comparatively study of % yield and reaction time in different method and solvents is described. The structure characterization of synthesized compound was carried out by 1H NMR, IR and mass spectrometry. From the result, it is observed that the microwave irradiation method is superior than the other two method to the synthesis of Benzo[1,3] oxazine-2,4-dione derivatives. [ABSTRACT FROM AUTHOR]

Background: Frequently isolated in clinical settings and environmental sources, members of the Mycobacterium aviumintracellulare complex (MAIC) comprising genetically distinct species and subtypes holds significant agents responsible for opportunistic infections. The work is focused on molecular identification of a novel Mycobacterium species within the MAIC isolated from an aged dog suffering from pulmonary infection in a rural area of Meghalaya, India. Methods & Materials: The dog's nasal swabs were collected and after decontamination inoculated into Lowenstein Jensen Media. Since phenotypic characteristics were inconclusive and time consuming, molecular analysis was decisively adopted for the study. Genus confirmation was done on the basis of hsp65 gene amplification which was then subjected to PRA (PCR-Restriction Enzyme Pattern Analysis) using enzymes BstEII and HaeIII. To further speciate the isolate and determine its phylogenetic status, two important additional housekeeping genes rpoB, and 16S rRNA gene were included. Results: At the end of three months a smooth, single, nonpigmented colony was observed showing strong acid fast bacilli and amplified the genus-specific hsp65 gene. PRA-hsp65 profiles could only infer the isolate to be a Non-tuberculous Mycobacteria (NTM). The dog was instantly prescribed anti-tuberculosis drug and is currently undergoing therapy. BLAST analysis in NCBI exhibited no homology in the three genes with comparatively low similarity cut-offs. The 16S rRNA gene depicted 99% closeness to the reference strain of M. yongonense, rpoB gene showed 97% similarity to M. Indicus pranii, while non-MAIC member M. genevense was the closest hit for hsp65 with 97% similarity. All the three genes were individually subjected to phylogenetic analysis in MEGA6 taking together the reference ATCC sequences of MAIC and the closest BLAST hits which also presented similar ambiguous output with no conclusion on the isolate's species. Concatenation of the three sequences ultimately presented higher lucid discrimination and determined the isolate (HNP-1) as a novel entity within the MAIC (Figure 1). Conclusion: Phylogenetic analysis indicate the isolate as a distinct new member within MAIC with a unique clinical manifestation in the present dog's case by way of being less lethal as compared to other reported cases. [ABSTRACT FROM AUTHOR]