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The Methods of Choice for Extracellular Vesicles (EVs) Characterization
2017
In recent years, extracellular vesicles (EVs) have become a subject of intense study. These membrane-enclosed spherical structures are secreted by almost every cell type and are engaged in the transport of cellular content (cargo) from parental to target cells. The impact of EVs transfer has been observed in many vital cellular processes including cell-to-cell communication and immune response modulation; thus, a fast and precise characterization of EVs may be relevant for both scientific and diagnostic purposes. In this review, the most popular analytical techniques used in EVs studies are presented with the emphasis on exosomes and microvesicles characterization.
Journal Article
Green Synthesized ZnO Nanoparticles Mediated by Mentha Spicata Extract Induce Plant Systemic Resistance against Tobacco Mosaic Virus
2020
Globally, plant viral infection is one of the most difficult challenges of food security, where considerable losses in crop production occur. Nanoparticles are an effective control agent against numerous plant pathogens. However, there is limited knowledge concerning their effects against viral infection. In the present study, the green synthesis of zinc oxide nanoparticles (ZnO NPs) using aqueous leaf extract of Mentha spicata was achieved. X-ray diffraction patterns confirmed the crystalline nature of the prepared ZnO NPs. Dynamic light scattering and scanning electron microscopy analyses revealed that the resultant ZnO NPs were spherical in shape with a particle size ranged from 11 to 88 nm. Fourier transmission infrared spectroscopy detected different functional groups, capping and stability agents, and showed Zn-O bond within wavenumber of 487 cm−1. Under greenhouse conditions, the antiviral activity of biological synthesized ZnO NPs (100 µg/mL) against Tobacco mosaic virus (TMV) was evaluated. The double foliar application of the prepared ZnO NPs, 24 h before and 24 h after TMV-inoculation, was the most effective treatment that showed a 90.21% reduction of viral accumulation level and disease severity. Additionally, the transcriptional levels of PAL, PR-1 (salicylic acid marker gene), CHS, and POD genes were induced and up-regulated in all ZnO NPs treated plants. Notably, the results exhibited that aqueous extract of Mentha spicata was an effective reducing agent for the green synthesis of ZnO NPs, which showed significant antiviral activity. Finally, the detected protective and curative activity of ZnO NPs against TMV can encourage us to recommend its application for plant viral disease management. To our knowledge, this is the first study describing the antiviral activity of the green synthesized ZnO NPs.
Journal Article
Improved Genome Sequence of Wild Emmer Wheat Zavitan with the Aid of Optical Maps
2019
Wild emmer (Triticum turgidum ssp. dicoccoides) is the progenitor of all modern cultivated tetraploid wheat. Its genome is large (> 10 Gb) and contains over 80% repeated sequences. The successful whole-genome-shotgun assembly of the wild emmer (accession Zavitan) genome sequence (WEW_v1.0) was an important milestone for wheat genomics. In an effort to improve this assembly, an optical map of accession Zavitan was constructed using Bionano Direct Label and Stain (DLS) technology. The map spanned 10.4 Gb. This map and another map produced earlier by us with the Bionano’s Nick Label Repair and Stain (NLRS) technology were used to improve the current wild emmer assembly. The WEW_v1.0 assembly consisted of 151,912 scaffolds. Of them, 3,102 could be confidently aligned on the optical maps. Forty-seven were chimeric. They were disjoined and new scaffolds were assembled with the aid of the optical maps. The total number of scaffolds was reduced from 151,912 to 149,252 and N50 increased from 6.96 Mb to 72.63 Mb. Of the 149,252 scaffolds, 485 scaffolds, which accounted for 97% of the total genome length, were aligned and oriented on genetic maps, and new WEW_v2.0 pseudomolecules were constructed. The new pseudomolecules included 333 scaffolds (68.51 Mb) which were originally unassigned, 226 scaffolds (554.84 Mb) were placed into new locations, and 332 scaffolds (394.83 Mb) were re-oriented. The improved wild emmer genome assembly is an important resource for understanding genomic modification that occurred by domestication.
Journal Article
Measuring ecosystem services based on government intentions for future land use in Hubei Province: implications for sustainable landscape management
2021
ContextExploring how ecosystem service values (ESV) are likely to change based on government intentions to develop and protect land is essential for sustainable landscape management.Objectives(1) Simulate land use change under future baseline (BAS), resource consumption (CON), and resource protection (PRO) scenarios, based on forecasted land expropriation prices implemented by the government of Hubei Province. (2) Measure changes in ecosystem services influenced by future land use. (3) Provide sustainable landscape management strategies to control the risk of ecosystem service loss.MethodsThis research couples Computable General Equilibrium of Land Use Change and Dynamics of Land System (CGELUC-DLS) models to simulate land use changes and calculated ESV using the equivalent factor method.Results(1) Predicted areas of cultivated land, forest, and grassland throughout Hubei Province declined under the three scenarios between 2015 and 2025. (2) Compared with 2015, equivalent values per unit area of ecosystem services (ESVe) decreased by 2.27%, 4.01%, and 1.67% in 2025 under BAS, CON, and PRO scenarios, respectively. The future trend in ESVe reduction across western Hubei Province did abate in the PRO scenario. (3) Reasonably adjusting land expropriation prices is a regulatory approach that can serve to strengthen sustainable landscape management in China.ConclusionsESV will inevitably decline in the future due to continuous land use changes across Hubei Province. The government should implement diversified strategies to control ecosystem services loss, including adjusting land expropriation prices, adopting regional differentiated management strategies, and implementing intensive but sustainable land use policies.
Journal Article
Cancer Diagnosis through SERS and Other Related Techniques
by
Alvarez-Puebla, Ramon A.
,
Blanco-Formoso, Maria
in
Biomarkers
,
Biomarkers, Tumor - blood
,
Biomarkers, Tumor - urine
2020
Cancer heterogeneity increasingly requires ultrasensitive techniques that allow early diagnosis for personalized treatment. In addition, they should preferably be non-invasive tools that do not damage surrounding tissues or contribute to body toxicity. In this context, liquid biopsy of biological samples such as urine, blood, or saliva represents an ideal approximation of what is happening in real time in the affected tissues. Plasmonic nanoparticles are emerging as an alternative or complement to current diagnostic techniques, being able to detect and quantify novel biomarkers such as specific peptides and proteins, microRNA, circulating tumor DNA and cells, and exosomes. Here, we review the latest ideas focusing on the use of plasmonic nanoparticles in coded and label-free surface-enhanced Raman scattering (SERS) spectroscopy. Moreover, surface plasmon resonance (SPR) spectroscopy, colorimetric assays, dynamic light scattering (DLS) spectroscopy, mass spectrometry or total internal reflection fluorescence (TIRF) microscopy among others are briefly examined in order to highlight the potential and versatility of plasmonics.
Journal Article
The Antimicrobial Activities of Silver Nanoparticles from Aqueous Extract of Grape Seeds against Pathogenic Bacteria and Fungi
by
Al-Askar, Abdulaziz A.
,
Alharbi, Raedah I.
,
Al-Otibi, Fatimah
in
Antifungal agents
,
Antimicrobial agents
,
Aqueous solutions
2021
Grape seed extract (GSE) is a natural source of polyphenolic compounds and secondary metabolites, which have been tested for their possible antimicrobial activities. In the current study, we tested the antibacterial and antifungal activities of aqueous GSE and the biosynthesized silver nanoparticles loaded with GSE (GSE-AgNPs) against different pathogens. The biosynthesized GSE-AgNPs were assessed by UV spectroscopy, dynamic light scattering (DLS), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), and gas chromatography/mass spectrometry (GC/MS). The antimicrobial activities were assessed against different bacterial and fungal species. DLS analysis showed that GSE-AgNPs had a Z-Average of 91.89 nm while UV spectroscopy showed that GSE-AgNPs had the highest absorbance at a wavelength of ~415 nm. FTIR analysis revealed that both of GSE and GSE-AgNPs consisted of different functional groups, such as hydroxyl, alkenes, alkyne, and aromatic rings. Both FE-SEM and TEM showed that GSE-AgNPs had larger sizes and rough surfaces than GSE and AgNO3. The results showed significant antimicrobial activities of GSE-AgNPs against all tested species, unlike GSE, which had weaker and limited effects. More studies are needed to investigate the other antimicrobial activities of GSE.
Journal Article
The Role of Mucin in the Toxicological Impact of Polystyrene Nanoparticles
by
Medina, Carlos
,
Tajber, Lidia
,
Inkielewicz-Stepniak, Iwona
in
Apoptosis
,
Biotechnology
,
Diagnostic systems
2018
The development of novel oral drug delivery systems is an expanding area of research and both new approaches for improving their efficacy and the investigation of their potential toxicological effect are crucial and should be performed in parallel. Polystyrene nanoparticles (NPs) have been used for the production of diagnostic and therapeutic nanosystems, are widely used in food packaging, and have also served as models for investigating NPs interactions with biological systems. The mucous gel layer that covers the epithelium of the gastrointestinal system is a complex barrier-exchange system that it is mainly constituted by mucin and it constitutes the first physical barrier encountered after ingestion. In this study, we aimed to investigate the effect of polystyrene NPs on mucin and its potential role during NP–cell interactions. For this purpose, we evaluated the interaction of polystyrene NPs with mucin in dispersion by dynamic light scattering and with a deposited layer of mucin using a quartz crystal microbalance with dissipation technology. Next, we measured cell viability and the apoptotic state of three enterocyte-like cell lines that differ in their ability to produce mucin, after their exposure to the NPs. Positive charged NPs showed the ability to strongly interact and aggregate mucin in our model. Positive NPs affected cell viability and induced apoptosis in all cell lines independently of their ability of produce mucin.
Journal Article
Analytical Platform for Monitoring Aggregation of Monoclonal Antibody Therapeutics
2019
PurposeTo develop an analytical platform for the estimation as well as characterization of aggregates over the complete size spectrum (from invisible monomer to visible precipitates).MethodsTwo mAb samples were incubated at 30°C in different buffer systems of protein A chromatography for observing degradation due to aggregation. The aggregation in these samples was quantified by size exclusion chromatography (SEC), dynamic light scattering (DLS), and micro flow imaging (MFI).ResultsThe results obtained from various characterization tools were analysed in various size ranges - size exclusion chromatography (SEC) (1 nm - 25 nm), dynamic light scattering (DLS) (10 nm - 5 μm), and micro flow imaging (MFI) (2 μm - 300 μm). Since each characterization tool covers a particular size range, data from multiple tools was collected in the “handover” regions to demonstrate accuracy of the platform.ConclusionsBased on the observations from the experiments, an analytical platform has been proposed covering the whole size spectrum that would be of utility to those engaged in formulation development as well as other aspects related to stability of biotherapeutic products.
Journal Article
Green synthesis of silver nanoparticles and its antibacterial activity using fungus Talaromyces purpureogenus isolated from Taxus baccata Linn
2022
The present study is focused on the synthesis of silver nanoparticles (AgNPs) utilizing endophytic fungus Talaromyces purpureogenus, isolated from Taxus baccata Linn. Extracellular extract of Talaromyces purpureogenus has shown occurrence of secondary metabolites viz. terpenoids and phenols. Gas chromatography-mass spectroscopy analysis showed the presence of 16 compounds. Techniques like Ultraviolet–visible spectroscopy, Fourier transform infrared spectroscopy, dynamic light scattering, field emission gun scanning electron microscopy, high-resolution transmission electron microscopy, energy-dispersive X-ray spectroscopy and X-ray diffraction were employed to characterize the synthesized AgNPs. UV–Vis spectroscopy showed sharp peaks at 380–470 nm which indicates the presence of metallic silver. FTIR analysis showed the presence of various functional groups like phenols, hydroxyl groups, and primary amines. In DLS, Z-average size and PdI of synthesized AgNPs were 240.2 r.nm and 0.720 respectively, with zeta potential − 19.6 mV. In FEG-SEM and HRTEM the spherical AgNPs showed diameter in the range of 30–60 nm. In EDS analysis the weight percent of Ag is 67.26% and atomic percent is 43.13%. From XRD analysis the size of AgNPs was found to be 49.3 nm with face-centered cubic crystalline nature of fungal synthesized AgNPs. These nanoparticles have shown significant antibacterial activity against tested strains viz. Listeria monocytogenes (13 ± 0.29 mm), Escherichia coli (17 ± 0.14 mm), Shigella dysenteriae (18 ± 0.21 mm) and Salmonella typhi (14 ± 0.13 mm). These synthesized AgNPs have shown effective free radical scavenging activity against 2,2′-diphenyl-1-picrylhydrazyl. The present study showed that the endophytic fungus Talaromyces purpureogenus can be used as a prominent source to synthesize AgNPs by using biological, ecofriendly, and in a non-toxic way accompanied by antibacterial and antioxidant properties which further can reduce the harvesting pressure faced by Taxus baccata.
Journal Article
Biosynthesis of zinc oxide nanoparticles using leaf extract of Calotropis gigantea: characterization and its evaluation on tree seedling growth in nursery stage
by
Chaudhuri, Sadhan Kumar
,
Malodia, Lalit
in
Absorption spectra
,
Atomic force microscopy
,
Biomolecules
2017
Green synthesis of zinc oxide nanoparticles was carried out using
Calotropis
leaf extract with zinc acetate salt in the presence of 2 M NaOH. The combination of 200 mM zinc acetate salt and 15 ml of leaf extract was ideal for the synthesis of less than 20 nm size of highly monodisperse crystalline nanoparticles. Synthesized nanoparticles were characterized through UV–Vis spectroscopy, dynamic light scattering (DLS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), EDX (energy dispersive X-ray), and AFM (atomic force microscopy). Effects of biogenic zinc oxide (ZnO) nanoparticles on growth and development of tree seedlings in nursery stage were studied in open-air trenches. The UV–Vis absorption maxima showed peak near 350 nm, which is characteristic of ZnO nanoparticles. DLS data showed that single peak is at 11 nm (100%) and Polydispersity Index is 0.245. XRD analysis showed that these are highly crystalline ZnO nanoparticles having an average size of 10 nm. FTIR spectra were recorded to identify the biomolecules involved in the synthesis process, which showed absorption bands at 4307, 3390, 2825, 871, 439, and 420 cm
−1
. SEM images showed that the particles were spherical in nature. The presence of zinc and oxygen was confirmed by EDX and the atomic % of zinc and oxygen were 33.31 and 68.69, respectively. 2D and 3D images of ZnO nanoparticles were obtained by AFM studies, which indicated that these are monodisperse having size ranges between 1.5 and 8.5 nm. Significant enhancement of growth was observed in Neem (
Azadirachta indica
), Karanj (
Pongamia pinnata
), and Milkwood-pine (
Alstonia scholaris
) seedlings in foliar spraying ZnO nanoparticles to nursery stage of tree seedlings. Out of the three treated saplings,
Alstonia scholaris
showed maximum height development.
Journal Article