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Interaction of Ras Binding Domain (RBD) by chemotherapeutic zinc oxide nanoparticles: Progress towards RAS pathway protein interference.

ABSTRACT: Zinc oxide (ZnO) NP is considered as a nanoscale chemotherapeutic. Thus, the drug delivery of this inorganic NP is of considerable importance. Ras mutations are common in cancer and the activation of this signaling pathway is a hallmark in carcinoma, melanoma and many other aggressive malignancies. Thus, here we examined the binding and delivery of Ras binding domain (RBD), a model cancer-relevant protein and effector of Ras by ZnO NP. Shifts in zeta potential in water, PBS, DMEM and DMEM supplemented with FBS supported NP interaction to RBD. Fluorescence quenching of the NP was concentration-dependent for RBD, Stern-Volmer analysis of this data was used to estimate binding strength which was significant for ZnO-RBD (Kd < 10-5). ZnO NP interaction to RBD was further confirmed by pull-down assay demonstrated by SDS-PAGE analysis. The ability of ZnO NP to inhibit 3-D tumor spheroid was demonstrated in HeLa cell spheroids-the ZnO NP breaking apart these structures revealing a significant (>50%) zone of killing as shown by light and fluorescence microscopy after intra-vital staining. ZnO 100 nm was superior to ZnO 14 nm in terms of anticancer activity. When bound to ZnO NP, the anticancer activity of RBD was enhanced. These data indicate the potential diagnostic application or therapeutic activity of RBD-NP complexes in vivo which demands further investigation.

SUBMITTER: Mathew EN

PROVIDER: S-EPMC7744048 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Interaction of Ras Binding Domain (RBD) by chemotherapeutic zinc oxide nanoparticles: Progress towards RAS pathway protein interference.

Mathew Elza Neelima EN Hurst Miranda N MN Wang Baolin B Murthy Vaibhav V Zhang Yuntao Y DeLong Robert K RK

PloS one 20201216 12

Zinc oxide (ZnO) NP is considered as a nanoscale chemotherapeutic. Thus, the drug delivery of this inorganic NP is of considerable importance. Ras mutations are common in cancer and the activation of this signaling pathway is a hallmark in carcinoma, melanoma and many other aggressive malignancies. Thus, here we examined the binding and delivery of Ras binding domain (RBD), a model cancer-relevant protein and effector of Ras by ZnO NP. Shifts in zeta potential in water, PBS, DMEM and DMEM supple ...[more]

PMID: 33326476

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Project description:BackgroundSeed germination is a critical stage in plant life, and recent practices use nanomaterials for the improvement of plant seed germination indices. This study was conducted to assess the effect of laboratory prepared zinc oxide nanoparticles on the physiological and biochemical changes of lettuce seeds.MethodsLettuce seeds were soaked in a suspension of moderately polydisperse zinc oxide nanoparticles at two different concentrations (25 ppm or 50 ppm) and shaken for 3 h at 25 °C. Seeds treatment was followed subsequently by two to three days drying at ambient conditions. Treated seeds were stored for 3-4 weeks, at ambient conditions and then tested for germination in petri dishes. Germination was observed on daily basis and seedling length was measured. After imbibition and before the start of the visible germination, seeds were examined for topography and surface analysis using the scanning electron microscope and zinc uptake was measured by using the atomic absorption spectrometry and the energy dispersive X-ray. The pattern of mobilization of biomolecules was analyzed to detect any differences among different seed groups.ResultsThere was no loss of viability for the nanoparticles treated seeds. Indeed their germination was enhanced and their biomass increased. The activated performance of the nanoparticles imbibed seeds has been found to be correlated with an increased level of Zn inside lettuce seeds. The recorded measurements show a significant enhancement of seedling length. Interaction of zinc oxide nanoparticles with lettuce seeds mediates a variation in the biochemical processes. Changes detected in treated seeds were as following: reduced levels of the total carbohydrates (including simple saccharides and polysaccharides), higher capacity of protein synthesis, an elevated level of starch as well as an increased activity of antioxidant enzymes.Discussion and conclusionLettuce seeds primed with ZnO nanoparticles were found not only to maintain seed viability but even to exhibit a detectable level of germination enhancement compared to the control seeds. Overall, the promoted response of lettuce seeds during early stages of seed growth is encouraging for the application of ZnO NPs for seed priming for better germination indices.

Project description:BackgroundThe high doses of zinc oxide (ZnO) administered orally to piglets for the prevention of diarrhea and increase of growth rate can contaminate pig farms and the surrounding environment. Therefore, there is a need to find a replacement of high doses of dietary ZnO with an equally effective alternative. In the present study, the effect of two formulations of zinc phosphate-based nanoparticles (ZnA and ZnC NPs) on growth performance, intestinal microbiota, antioxidant status, and intestinal and liver morphology was evaluated. A total of 100 weaned piglets were randomly divided into 10 equal groups with the base diet (control) or the base diet supplemented with ZnA, ZnC, or ZnO at concentrations 500, 1000, and 2000?mg Zn per kilogram of diet. Supplements were given to animals for 10?days. Fecal samples were collected on day 0, 5, 10 and 20. At the end of the treatment (day 10), three piglets from each group were sacrificed and analyzed.ResultsComparing to that of control, the significantly higher piglet weight gain was observed in all piglet groups fed with ZnA (P?<?0.05). Differences in the total aerobic bacteria and coliform counts in piglet feces after NPs supplementation compared to that of control and ZnO groups were also found (P?<?0.05). The majority of aerobic culturable bacteria from the feces represented Escherichia (28.57-47.62%), Enterococcus (3.85-35.71%), and Streptococcus (3.70-42.31%) spp. A total of 542 Escherichia coli isolates were screened for the virulence genes STa, STb, Stx2, F4, and F18. The substantial occurrence of E. coli virulence factors was found on day 5, mainly in fimbrillary antigen and thermostable toxins, except for piglets fed by ZnC. Zn treatment decreased Zn blood levels in piglets fed with ZnO and ZnA (500?mg/kg) and increased in ZnC (2000?mg/kg) compared to that of control (P?<?0.05). The antioxidant status of piglets was affected only by ZnA. While some changes in the liver and the intestinal morphology of piglets with NPs were observed, none were serious as reflected by the normal health status and increased weigh gain performance.ConclusionsOur results indicate that ZnA NPs have a positive effect on the piglet growth performance even at the lowest concentration. The prevalence of E. coli virulence factors was lowest in pigs supplemented with ZnC. Zinc phosphate-based nanoparticles may be an effective alternative to ZnO.

Dataset Information

Interaction of Ras Binding Domain (RBD) by chemotherapeutic zinc oxide nanoparticles: Progress towards RAS pathway protein interference.

Publications

Interaction of Ras Binding Domain (RBD) by chemotherapeutic zinc oxide nanoparticles: Progress towards RAS pathway protein interference.

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