This work examines the biological activity of essential oils of Cinnamomum camphora leaves, C. glaucescens fruit, and C. tamala root from Nepal. The oils were screened for phytotoxic activity against lettuce and perennial ryegrass, brine shrimp lethality, and antibacterial, antifungal, cytotoxic, insecticidal, and nematicidal activities. C. camphora leaf essential oil was phytotoxic to lettuce, antifungal to Aspergillus niger, and insecticidal, particularly toward midge and butterfly larvae, fruit flies, and fire ants. C. camphora oil was also toxic to brine shrimp and human breast tumor cells. C. glaucescens fruit essential oil showed notable nematicidal activity, as well as termiticidal and mosquito larvicidal activity. The root essential oil of C. tamala was toxic to mosquito larvae and fire ants.

USP9X is a conserved deubiquitinase (DUB) that regulates multiple cellular processes. Dysregulation of USP9X has been linked to cancers and X-linked intellectual disability. Here, we report the crystal structure of the USP9X catalytic domain at 2.5-Å resolution. The structure reveals a canonical USP-fold comprised of fingers, palm, and thumb subdomains, as well as an unusual β-hairpin insertion. The catalytic triad of USP9X is aligned in an active configuration. USP9X is exclusively active against ubiquitin (Ub) but not Ub-like modifiers. Cleavage assays with di-, tri-, and tetraUb chains show that the USP9X catalytic domain has a clear preference for K11-, followed by K63-, K48-, and K6-linked polyUb chains. Using a set of activity-based diUb and triUb probes (ABPs), we demonstrate that the USP9X catalytic domain has an exo-cleavage preference for K48- and endo-cleavage preference for K11-linked polyUb chains. The structure model and biochemical data suggest that the USP9X catalytic domain harbors three Ub binding sites, and a zinc finger in the fingers subdomain and the β-hairpin insertion both play important roles in polyUb chain processing and linkage specificity. Furthermore, unexpected labeling of a secondary, noncatalytic cysteine located on a blocking loop adjacent to the catalytic site by K11-diUb ABP implicates a previously unreported mechanism of polyUb chain recognition. The structural features of USP9X revealed in our study are critical for understanding its DUB activity. The new Ub-based ABPs form a set of valuable tools to understand polyUb chain processing by the cysteine protease class of DUBs.

Glyphosate [N‐(phosphonomethyl) glycine] is a broad‐ spectrum systemic herbicide. Because of the ever‐increasing application of glyphosate in agricultural fields and detection of its presence in soils and environmental waters, there is a growing concern that it may potentially harm animal and human populations and the environment. In this study, we determined the mechanism and reaction pathway of birnessite‐catalyzed degradation of glyphosate by using one‐dimensional (1H, 13C, and 31P) and two‐dimensional homo‐ and hetero‐nuclear NMR correlation spectroscopies. The NMR results showed the presence of several degradation products including orthophosphate, aminomethylphosphonic acid (AMPA), glycine, and sarcosine. Two‐dimensional NMR and correlation spectroscopies confirmed the identity of products and contaminants in the reactions. Our results show that the glyphosate degradation reaction is very fast, with the majority of reaction products formed within 1 min of reaction. Degradation depends linearly on the birnessite concentration, indicating sorption as the prerequisite for glyphosate degradation. Based on the reaction kinetics and NMR data, we propose two glyphosate degradation pathways: one starts with C–P bond cleavage and the other with C–N bond cleavage. Interestingly, the dominance of one pathway over the other was found to vary with the birnessite/glyphosate ratio. This information could be exploited further to identify conditions under which glyphosate degradation occurs primarily under the C–P bond cleavage pathway because this pathway does not produce AMPA, a compound with a longer half‐life than glyphosate and with known animal and plant toxicity.

Degradation of glyphosate in the presence of manganese oxide and UV light was analyzed using phosphate oxygen isotope ratios and density function theory (DFT). The preference of C-P or C-N bond cleavage was found to vary with changing glyphosate/manganese oxide ratios, indicating the potential role of sorption-induced conformational changes on the composition of intermediate degradation products. Isotope data confirmed that one oxygen atom derived solely from water was incorporated into the released phosphate during glyphosate degradation, and this might suggest similar nucleophilic substitution at P centers and C-P bond cleavage both in manganese oxide- and UV light-mediated degradation. The DFT results reveal that the C-P bond could be cleaved by water, OH or OH, with the energy barrier opposing bond dissociation being lowest in the presence of the radical species, and that C-N bond cleavage is favored by the formation of both nitrogen- and carbon-centered radicals. Overall, these results highlight the factors controlling the dominance of C-P or C-N bond cleavage that determines the composition of intermediate/final products and ultimately the degradation pathway.

The essential oils from the leaves of Artemisia dubia, A. indica, and A. vulgaris growing wild in Nepal were obtained by hydrodistillation and analyzed by GC-MS. The major components in A. dubia oil were chrysanthenone (29.0%), coumarin (18.3%), and camphor (16.4%). A. indica oil was dominated by ascaridole (15.4%), isoascaridole (9.9%), trans-p-mentha-2,8-dien-1-ol (9.7%), and trans-verbenol (8.4%). The essential oil of Nepalese A. vulgaris was rich in α-thujone (30.5%), 1,8-cineole (12.4%), and camphor (10.3%). The essential oils were screened for phytotoxic activity against Lactuca sativa (lettuce) and Lolium perenne (perennial ryegrass) using both seed germination and seedling growth, and all three Artemisia oils exhibited notable allelopathic activity. A. dubia oil showed in-vitro cytotoxic activity on MCF-7 cells (100% kill at 100 μg/mL) and was also marginally antifungal against Aspergillus niger (MIC = 313 μg/mL). DFT calculations (B3LYP/6-31G*) revealed thermal decomposition of ascaridole to be energetically accessible at hydrodistillation and GC conditions, but these are spin-forbidden processes. If decomposition does occur, it likely proceeds by way of homolytic peroxide bond cleavage rather than retro-Diels-Alder elimination of molecular oxygen.