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Search for "protein" in Full Text gives 382 result(s) in Beilstein Journal of Nanotechnology. Showing first 200.

Alcohol-perturbed self-assembly of the tobacco mosaic virus coat protein

  • Ismael Abu-Baker and
  • Amy Szuchmacher Blum

Beilstein J. Nanotechnol. 2022, 13, 355–362, doi:10.3762/bjnano.13.30

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  • Ismael Abu-Baker Amy Szuchmacher Blum Department of Chemistry, McGill University, Montréal, Québec, Canada 10.3762/bjnano.13.30 Abstract The self-assembly of the tobacco mosaic virus coat protein is significantly altered in alcohol–water mixtures. Alcohol cosolvents stabilize the disk aggregate
  • alcohol having the strongest effect and methanol the weakest. We discuss several effects that may contribute to preventing the proteinprotein interactions between disks that are necessary to form helical rods. Keywords: alcohol; hydrophobic effect; protein assembly; self-assembly; tobacco mosaic virus
  • applications [7][8]. Many viruses and VLPs form monodisperse particles due to the natural capsid protein symmetry and inter-subunit interactions, as well as interactions with encapsidated genetic material [9][10]. Viruses can be obtained in high yields by propagation in host organisms, and viral capsid
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Published 01 Apr 2022

Systematic studies into uniform synthetic protein nanoparticles

  • Nahal Habibi,
  • Ava Mauser,
  • Jeffery E. Raymond and
  • Joerg Lahann

Beilstein J. Nanotechnol. 2022, 13, 274–283, doi:10.3762/bjnano.13.22

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  • , or monodispersity. Electrohydrodynamic (EHD) jetting is a probate method to formulate synthetic protein nanoparticles (SPNPs), but a systematic understanding of the influence of crucial processing parameters, such as protein composition, on nanoparticle morphologies is still missing. Here, we address
  • this knowledge gap by evaluating formulation trends in SPNPs prepared by EHD jetting based on a series of carrier proteins and protein blends (hemoglobin, transferrin, mucin, or insulin). In general, blended SPNPs presented uniform populations with minimum diameters between 43 and 65 nm. Size
  • , blended SPNPs displayed higher circularity and lower anisotropy, as compared to single-protein SPNPs. Secondary statistical analysis indicated that blended SPNPs generally present combined features of their constituents, with some properties driven by the dominant protein constituent. Our study suggests
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Published 28 Feb 2022

Photothermal ablation of murine melanomas by Fe3O4 nanoparticle clusters

  • Xue Wang,
  • Lili Xuan and
  • Ying Pan

Beilstein J. Nanotechnol. 2022, 13, 255–264, doi:10.3762/bjnano.13.20

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  • experimental approaches including semi-quantitative PCR, western blot and immunohistochemistry, the heat shock protein HSP70 to be upregulated in response to PTT, suggesting this chaperone protein could be a plausible underlying mechanism for the observed therapeutic outcome. Altogether, our results highlight
  • the promise of Fe3O4 NPCs as a new PTT option to treat melanoma. Keywords: Fe3O4 nanoparticle clusters; heat shock protein 70; melanoma; near infrared; photothermal therapy; Introduction The global incidence of melanoma, one of the deadliest forms of cancer, has kept increasing annually over the
  • NIR irradiation. HSP70 is frequently activated in response to heat stress, and earlier studies have found that it promotes apoptosis at least by eliciting antitumor immune responses [21]. This chaperone protein has been shown in our earlier study to be involved in mediating the photothermal effects of
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Published 22 Feb 2022

Engineered titania nanomaterials in advanced clinical applications

  • Padmavati Sahare,
  • Paulina Govea Alvarez,
  • Juan Manual Sanchez Yanez,
  • Gabriel Luna-Bárcenas,
  • Samik Chakraborty,
  • Sujay Paul and
  • Miriam Estevez

Beilstein J. Nanotechnol. 2022, 13, 201–218, doi:10.3762/bjnano.13.15

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  • excretion [33]. When a TiO2 nanomaterial circulates through the body, certain biomolecules (such as proteins, phospholipids, or DNA contained in biological fluids or present in living cells) get adsorbed onto the surface of it very quickly, which is termed as “protein corona (PC)” formation. This protein
  • integrin clustering and focal adhesion development. In this context, Chen et al. employed the adsorption of functional proteins (bone morphogenetic protein 2 and sclerostin antibody) to modify TiO2 nanotube arrays to repair bone fractures [35]. The PC alters biodistribution, biological identity and
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Published 14 Feb 2022

A comprehensive review on electrospun nanohybrid membranes for wastewater treatment

  • Senuri Kumarage,
  • Imalka Munaweera and
  • Nilwala Kottegoda

Beilstein J. Nanotechnol. 2022, 13, 137–159, doi:10.3762/bjnano.13.10

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  • hydrophobic PVDF membrane to a hydrophilic membrane with a water contact angle of 22.72°. The nanohybrid membrane showed a dye removal efficiency of 88.9% and an adsorption capacity of 72.6 mg/g for IC [78]. Huong et al. developed a waste protein-immobilized cationic dye removal membrane from PAN. In this
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Published 31 Jan 2022

Bacterial safety study of the production process of hemoglobin-based oxygen carriers

  • Axel Steffen,
  • Yu Xiong,
  • Radostina Georgieva,
  • Ulrich Kalus and
  • Hans Bäumler

Beilstein J. Nanotechnol. 2022, 13, 114–126, doi:10.3762/bjnano.13.8

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  • /bjnano.13.8 Abstract Hemoglobin microparticles (HbMP) produced with a three-step procedure, including coprecipitation of hemoglobin with manganese carbonate, protein cross-linking, and dissolution of the carbonate template were shown to be suitable for application as artificial oxygen carriers. First
  • the HbMP fabrication process to dissolve the manganese carbonate template to produce pure protein particles. EDTA is widely utilized in medical and biological applications. Because of its chelating properties, it is used as an anticoagulant in blood samples [27]. It has also long been used to
  • inhibit DNA synthesis in bacteria, and similar effects are also seen on RNA and protein syntheses [44][45]. In addition, glutaraldehyde particularly acts on the outer layers of E. coli and cross-links lipoproteins and proteins there as well. This fixation of bacteria prevents the bacterial cells from
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Published 24 Jan 2022

Theranostic potential of self-luminescent branched polyethyleneimine-coated superparamagnetic iron oxide nanoparticles

  • Rouhollah Khodadust,
  • Ozlem Unal and
  • Havva Yagci Acar

Beilstein J. Nanotechnol. 2022, 13, 82–95, doi:10.3762/bjnano.13.6

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  • was recorded using a UV–vis spectrophotometer (Shimadzu UV-3600 UV–vis–NIR spectrophotometer). A calibration curve was prepared with bovine serum albumin (BSA) at concentration values of 0, 1, 2, 4, 6, 8, and 10 µg/mL in PBS to enable a correlation between the measured absorbance and protein
  • to the outer cell surface (without activation of caspase 3) and also leakage of cytosolic lactate dehydrogenase (LDH) from the cells [55][56]. It has also been known that unmodified polyplexes can switch on the intrinsic apoptotic pathway by activating the box protein and therefore releasing the
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Published 18 Jan 2022

Biocompatibility and cytotoxicity in vitro of surface-functionalized drug-loaded spinel ferrite nanoparticles

  • Sadaf Mushtaq,
  • Khuram Shahzad,
  • Tariq Saeed,
  • Anwar Ul-Hamid,
  • Bilal Haider Abbasi,
  • Nafees Ahmad,
  • Waqas Khalid,
  • Muhammad Atif,
  • Zulqurnain Ali and
  • Rashda Abbasi

Beilstein J. Nanotechnol. 2021, 12, 1339–1364, doi:10.3762/bjnano.12.99

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  • potential values is the interaction between NPs and serum proteins present in DMEM [30]. In cell culture media, NPs agglomerate with serum proteins and are therefore recruited in cells via the protein corona effect, which increases the bioavailability of NPs by many folds [31]. PMA-coated samples have a
  • p53 is a tumor suppressor protein also known as the “guardian of the genome”. It is involved in downstream regulation of genes involved in apoptosis, DNA repair, and cell cycle arrest. Overexpression of p53 is triggered by stress stimuli, such as hypoxia, ROS, ionizing radiations, and carcinogens
  • efflux chemotherapeutic drugs, a phenomenon known as MDR. P-glycoprotein belongs to the ABCB1 family of ABC proteins and is involved in the efflux of doxorubicin, paclitaxel, vincristine, rhodamine, and etoposide [63]. Conversely, multidrug resistance-associated protein 1 (MRP1) is a member of the ABCC1
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Published 02 Dec 2021

Self-assembly of amino acids toward functional biomaterials

  • Huan Ren,
  • Lifang Wu,
  • Lina Tan,
  • Yanni Bao,
  • Yuchen Ma,
  • Yong Jin and
  • Qianli Zou

Beilstein J. Nanotechnol. 2021, 12, 1140–1150, doi:10.3762/bjnano.12.85

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  • function due to the collective behavior of aggregates [24]. However, protein and peptide assemblies exhibit disadvantages, such as difficulty in obtaining high quantities of materials, corresponding high costs, and in some cases polydispersity and purification limitations. When using proteins and peptides
  • as functional materials it can be difficult to arrange assembled nanostructures at both microscopic and macroscopic scales [10]. Also, fine manipulation of noncovalent interactions and corresponding peptide and protein nanostructures remains a huge challenge [24]. Amino acids are the major components
  • . However, using these simple building blocks to design attractive materials remains a challenge due to the simplicity of the amino acids [26]. Most articles about self-assembly focus on peptide and protein self-assembly [15][29]. Therefore, it is of great significance to systematically summarize the latest
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Published 12 Oct 2021

Use of nanosystems to improve the anticancer effects of curcumin

  • Andrea M. Araya-Sibaja,
  • Norma J. Salazar-López,
  • Krissia Wilhelm Romero,
  • José R. Vega-Baudrit,
  • J. Abraham Domínguez-Avila,
  • Carlos A. Velázquez Contreras,
  • Ramón E. Robles-Zepeda,
  • Mirtha Navarro-Hoyos and
  • Gustavo A. González-Aguilar

Beilstein J. Nanotechnol. 2021, 12, 1047–1062, doi:10.3762/bjnano.12.78

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  • -cell lung cancer through the regulation of metastasis-associated protein 1 (MTA1), which is linked to tumor aggressiveness and metastasis, while its modulating effect is related to the suppression of the Wnt/β-catenin pathway. Curcumin also blocks pancreatic cancer metastases through PI3K/Akt/NF-ĸB
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Published 15 Sep 2021

An overview of microneedle applications, materials, and fabrication methods

  • Zahra Faraji Rad,
  • Philip D. Prewett and
  • Graham J. Davies

Beilstein J. Nanotechnol. 2021, 12, 1034–1046, doi:10.3762/bjnano.12.77

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  • from cancer research (25% enhanced absorption of the protein cancer drug AvastinTM compared with conventional hypodermic delivery) [14]. Incorporation of drug-loaded nanoparticles in dissolving microneedles also shows promise for dose concentration, for example using the antimicrobial carvacrol (CAR
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Published 13 Sep 2021

The role of convolutional neural networks in scanning probe microscopy: a review

  • Ido Azuri,
  • Irit Rosenhek-Goldian,
  • Neta Regev-Rudzki,
  • Georg Fantner and
  • Sidney R. Cohen

Beilstein J. Nanotechnol. 2021, 12, 878–901, doi:10.3762/bjnano.12.66

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  • (segmentation) is used as a good reference for image analysis approaches, that is, cell counting or tracking, and protein localization [12]. The segmentation can be performed as a “semantic segmentation”, which classifies image pixels into specific categories, while “instance segmentation” additionally segments
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Published 13 Aug 2021

Comprehensive review on ultrasound-responsive theranostic nanomaterials: mechanisms, structures and medical applications

  • Sepand Tehrani Fateh,
  • Lida Moradi,
  • Elmira Kohan,
  • Michael R. Hamblin and
  • Amin Shiralizadeh Dezfuli

Beilstein J. Nanotechnol. 2021, 12, 808–862, doi:10.3762/bjnano.12.64

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  • ) gas-filled structures that are stabilized by a lipid, surfactant, protein, or polymer shell, whose stiffness or rigidity can affect the final outcome of the MBs upon exposure to US [71][72][73]. During the cavitation event, backscattered energy leads to the expanding and shrinking of the MBs, which
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Published 11 Aug 2021

Silver nanoparticles induce the cardiomyogenic differentiation of bone marrow derived mesenchymal stem cells via telomere length extension

  • Khosro Adibkia,
  • Ali Ehsani,
  • Asma Jodaei,
  • Ezzatollah Fathi,
  • Raheleh Farahzadi and
  • Mohammad Barzegar-Jalali

Beilstein J. Nanotechnol. 2021, 12, 786–797, doi:10.3762/bjnano.12.62

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  • absolute telomere length (TL) measurement, and gene and protein assessment of the cells, respectively. It was found that 2.5 µg/mL Ag-NPs caused elongation of the telomeres and altered VEGF, C-TnI, VWF, SMA, GATA-4, TERT, and cyclin D protein and gene expression in the cardiomyogenically differentiated BM
  • -MSCs. Also, there was a significant increase in the protein and gene expression of Wnt3 and β-catenin as main components of pathways. We concluded that Ag-NPs could change the in vitro expression of cardiac markers of BM-MSCs via the Wnt3/β-catenin signaling pathway. Keywords: bone marrow-derived MSCs
  • that Ag-NPs induced mESCs cell cycle arrest at the G1 and S phases through inhibition of the hyperphosphorylation of Retinoblastoma protein [12]. Kalishwaralal et al. reported that Ag-NPs could inhibit cell proliferation, cell viability, and cell migration through activating caspase-3 and suppressing
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Published 02 Aug 2021

Recent progress in magnetic applications for micro- and nanorobots

  • Ke Xu,
  • Shuang Xu and
  • Fanan Wei

Beilstein J. Nanotechnol. 2021, 12, 744–755, doi:10.3762/bjnano.12.58

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  • -assisted gene transfection. Magnetic MNRs containing nucleic acids were delivered to target cells by magnetic fields. This method could change gene function or protein expression, which is of great significance for future research on gene transfer and gene therapy. Table 1 summarizes the relevant
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Published 19 Jul 2021

Fate and transformation of silver nanoparticles in different biological conditions

  • Barbara Pem,
  • Marija Ćurlin,
  • Darija Domazet Jurašin,
  • Valerije Vrček,
  • Rinea Barbir,
  • Vedran Micek,
  • Raluca M. Fratila,
  • Jesus M. de la Fuente and
  • Ivana Vinković Vrček

Beilstein J. Nanotechnol. 2021, 12, 665–679, doi:10.3762/bjnano.12.53

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  • significantly affect AgNPs and determine their colloidal stability and cellular interactions as evidenced earlier [27][31][32][33]. In the acidic medium of the stomach, AgNPs both agglomerate and dissolve [15][26][34]. The transformation will likely be incomplete due to protein corona formation and short
  • behaviour can be attributed to the loss of electrostatic repulsion between particles due to the complexation with counter ions present in media with high ionic strength [8][47]. The presence of proteins prevented AgNP agglomeration in m(CCM+BSA), m(CYS+BSA), m(GSH+BSA) due to the formation of protein corona
  • AgNPs enter the body where they gain a protein corona, aggregate, and dissolve to Ag+. Ionic silver may precipitate in the anion-rich environment of different tissues, where Ag binds to S, resulting in nanocrystals. Aggregation and corona–NP destabilisation can also lead to precipitation. Whole blood
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Published 07 Jul 2021

The preparation temperature influences the physicochemical nature and activity of nanoceria

  • Robert A. Yokel,
  • Wendel Wohlleben,
  • Johannes Georg Keller,
  • Matthew L. Hancock,
  • Jason M. Unrine,
  • D. Allan Butterfield and
  • Eric A. Grulke

Beilstein J. Nanotechnol. 2021, 12, 525–540, doi:10.3762/bjnano.12.43

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  • from partial nanoceria dissolution led to less cellular protein carbonyl formation, attributed to an increased amount of surface Ce3+ species. Surface reactivity was greater for the solvothermally synthesized nanoceria, which had more Ce3+ species at the surface. The results show that temperature
  • of nanoceria dissolution on its biological identity was assessed as the protein carbonyl level in response to partially dissolved nanoceria. The catalytic potential was assessed using a reactivity assay. Experimental Materials NM-211 and NM-212 were obtained from the European Commission Joint
  • the presence of protease inhibitors (4 µg/mL leupeptin, 4 µg/mL pepstatin A, and 5 µg/mL aprotinin). Protein was measured using the bicinchoninic acid method. Protein carbonyls were determined as described in [45]. Briefly, samples were derivatized with 10 mM 2,4-dinitrophenylhydrazine solution in 12
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Published 04 Jun 2021

A review on nanostructured silver as a basic ingredient in medicine: physicochemical parameters and characterization

  • Gabriel M. Misirli,
  • Kishore Sridharan and
  • Shirley M. P. Abrantes

Beilstein J. Nanotechnol. 2021, 12, 440–461, doi:10.3762/bjnano.12.36

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  • human serum albumin (HSA), fibrinogen and immunoglobulin (IgG), metallothionein (MT), and ceruloplasmin (CP), forming a protein corona (PC) during silver homeostasis [121][122]. The PC is a highly dynamic system and its composition dynamically changes over time, undergoing various transformations until
  • ceruloplasmin (CP), both involved in metal homeostasis. They concluded that protein diversity stabilizes and controls the dissolution and transport of AgNPs. Understanding the interaction between AgNPs and corona proteins is essential to establish new in vivo studies with AgNPs [122]. The human immune system
  • aminotransferases and also in the hematological parameters were observed between the control and groups with burn wounds and between groups treated with AgNPs and groups with burn wounds. In addition, there were no significant differences in lipid peroxidation, carbonyl and reduced glutathione (GSH) protein levels
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Published 14 May 2021

The impact of molecular tumor profiling on the design strategies for targeting myeloid leukemia and EGFR/CD44-positive solid tumors

  • Nikola Geskovski,
  • Nadica Matevska-Geshkovska,
  • Simona Dimchevska Sazdovska,
  • Marija Glavas Dodov,
  • Kristina Mladenovska and
  • Katerina Goracinova

Beilstein J. Nanotechnol. 2021, 12, 375–401, doi:10.3762/bjnano.12.31

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  • introduced the possibility of employing similar strategies for passive targeting as for solid tumors, but these tumors also have much in common regarding the expression of specific molecules as viable targets for therapy and/or homing of NDDSs. For example, overexpression of the EGFR gene or protein kinase
  • %, from 50% in the pre-imatinib era [7]. However, imatinib and related TKIs (the second-generation TKIs dasatinib, bosutinib, nilotinib, and the third-generation ponatinib) are not exclusively specific for the BCR-ABL fusion protein, and may also affect normal c-ABL and other kinases such as c-KIT. This
  • oncogene or mutational events [16]. The clinically relevant mutations arise in the kinase domain of BCR-ABL1 and lead to impaired TKI activity, mainly by preventing the fusion protein from adopting the correct conformation required for specific binding. More than 40 different point mutations have been
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Published 29 Apr 2021

Characterization, bio-uptake and toxicity of polymer-coated silver nanoparticles and their interaction with human peripheral blood mononuclear cells

  • Sahar Pourhoseini,
  • Reilly T. Enos,
  • Angela E. Murphy,
  • Bo Cai and
  • Jamie R. Lead

Beilstein J. Nanotechnol. 2021, 12, 282–294, doi:10.3762/bjnano.12.23

Graphical Abstract
  • that aggregation played a large role in NP transformations, except at high concentrations, due to protection by the PVP, and at later times because of possible protein interactions in the RPMI medium [38][39]. This result is in agreement with previous studies that found that aggregation/agglomeration
  • ., passivation, or as separate particles) or are bound to proteins. These leads to the formation of secondary NPs, chloro complexes and protein interactions in the RPMI medium yielding a Ag compounds that have a molecular mass above the nominal value of the 3 kDa filter membrane, which affects filter permeation
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Published 24 Mar 2021

Differences in surface chemistry of iron oxide nanoparticles result in different routes of internalization

  • Barbora Svitkova,
  • Vlasta Zavisova,
  • Veronika Nemethova,
  • Martina Koneracka,
  • Miroslava Kretova,
  • Filip Razga,
  • Monika Ursinyova and
  • Alena Gabelova

Beilstein J. Nanotechnol. 2021, 12, 270–281, doi:10.3762/bjnano.12.22

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  • functionalization to increase their accumulation in the tumor mass [3]. Numerous coating moieties have been employed to modify the surface properties of MNPs [16]. Among them, polyethylene glycol (PEG), a non-degradable polyether of the monomer ethylene glycol, and bovine serum albumin (BSA), a versatile protein
  • through CavME or lipid rafts. Co-localization studies confirmed the entrapment of fluorescently labeled RITC-BSA-SO-MNPs in clathrin-coated vesicles in A549 cells stably expressing green fluorescent protein (GFP)-clathrin. The results indicate that tuning of the MNP surface chemistry can potentially
  • cellular uptake of nanoparticles. Therefore, initial experiments were focused on the expression of clathrin heavy chain (CLHC), dynamin (Dyn), caveolin-1 (Cav1), and its phosphorylated form (pCav1) in A549 cells. The expression of CLHC and Cav1 was determined at the protein (Supporting Information File 1
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Published 23 Mar 2021

The nanomorphology of cell surfaces of adhered osteoblasts

  • Christian Voelkner,
  • Mirco Wendt,
  • Regina Lange,
  • Max Ulbrich,
  • Martina Gruening,
  • Susanne Staehlke,
  • Barbara Nebe,
  • Ingo Barke and
  • Sylvia Speller

Beilstein J. Nanotechnol. 2021, 12, 242–256, doi:10.3762/bjnano.12.20

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  • current sensing and motion termination results in a larger error signal on hard materials than on soft materials. Thus, the comparison between ion current error images of living and fixed cells can provide insight into the protein content in the ruffle volume, because fixation leads to crosslinking of
  • stiffness of the ruffle structure, even when a harder (fixed) protein content is present. Note that for living cells such contrast is absent as can be seen in Figure 3c and Figure 3d. Figure 4a shows an example of a peripheral ruffle. Peripheral ruffles sometimes are attributed to loosened lamellipodia
  • membrane would correspond to 20 nm. Values in this regime are in line with protein-aided cellular membrane curvatures. For example, intracellular membrane tubes exhibit diameters of a few tens of nanometers, a maximum curvature is assumed to be generated via the Bin/ampiphysin/Rvs (BAR) protein and
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Published 12 Mar 2021

A review on the biological effects of nanomaterials on silkworm (Bombyx mori)

  • Sandra Senyo Fometu,
  • Guohua Wu,
  • Lin Ma and
  • Joan Shine Davids

Beilstein J. Nanotechnol. 2021, 12, 190–202, doi:10.3762/bjnano.12.15

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  • studying cellular toxicity, response to new drugs [78] and environmental pollution. Silkworm (Bombyx mori) is an invertebrate insect widely used as a model organism in life sciences [79] since it has diverse mutant strains, a complete genome sequenced, and a protein database available [80][81]. There are
  • suppressed the activity of the enzyme SOD and of the protein HSP 1. This, in turn, activated the oxidative stress pathway and the production of ROS, which have toxic effects on the silkworm digestive system. Effects of nanomaterial exposure on silkworm tissues and organs The application of nanotechnology in
  • the silk fibers, enhancing their mechanical properties and promoting silk protein crystallization. This is in line with the research report by Wu et al. [168], who stated that silkworm silk containing Cu exhibited a good tensile strength of 360 MPa and a strain of 38%, which is 89% and 36% higher than
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Published 12 Feb 2021

Imaging of SARS-CoV-2 infected Vero E6 cells by helium ion microscopy

  • Natalie Frese,
  • Patrick Schmerer,
  • Martin Wortmann,
  • Matthias Schürmann,
  • Matthias König,
  • Michael Westphal,
  • Friedemann Weber,
  • Holger Sudhoff and
  • Armin Gölzhäuser

Beilstein J. Nanotechnol. 2021, 12, 172–179, doi:10.3762/bjnano.12.13

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  • of micrometers, which demonstrates the high depth of field of HIM compared to SEM [37]. In image 3a3, at the surface of the cell, a cluster of virus particles seems to be bound to the cell membrane (arrow). We suggest that this resembles the particle clustering by host defense protein BST-2 as it was
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Published 02 Feb 2021

A review on the green and sustainable synthesis of silver nanoparticles and one-dimensional silver nanostructures

  • Sina Kaabipour and
  • Shohreh Hemmati

Beilstein J. Nanotechnol. 2021, 12, 102–136, doi:10.3762/bjnano.12.9

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  • respect to reaction conditions. FTIR results show that similar to bacteria, carbonyl, amide, and hydroxy groups corresponding to the cellular protein are responsible for the synthesis and stabilization of AgNPs [178][179]. The less non-pathogenic behavior of fungi and their faster synthesis rate suggest
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Published 25 Jan 2021
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