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• Multi-domain vaccines are valuable types of active cancer immunotherapy. • The vaccine constructed from three proteins: IL-13Rα2 , TNC , and PTPRZ-1. • Each domain was fused together with appropriate linkers. • Different strategies were applied to enhance immunogenicity of multi-domain vaccine. • The whole vaccine features were evaluated immunoinformatically. Glioblastoma multiform is the most common of primary malignant brain tumors in adults. Currently, surgical resection of the tumor mass, followed by adjuvant radiotherapy and chemotherapy are standard treatments for glioblastoma multiform but so far are not effective treatments. Thus, the development of a vaccine, as a safe and efficient strategy for prophylactic or therapeutic purposes against glioblastoma multiform is very necessary. The present study aimed to design the multi-domain vaccine for glioblastoma multiform. An in silico approach was used to select the most potent domains of proteins to induce the host's B- and T-cell immune response against glioblastoma multiform. IL-13Rα-2 (amino acid positions 27–144), TNC (amino acid positions 1900–2100), and PTPRZ-1(amino acid positions 731–884) were found to have potent inducible immune responses. So, we considered them for fusing with a linker A(EAAAK) 3 A to construct the multi-domain recombinant vaccine. The immuno-informatics analysis of the designed recombinant vaccine construct was performed to evaluate its efficacy. Although the designed recombinant vaccine construct did not show allergen property, its antigenicity was estimated at 0.78. The Physico-chemical properties of the recombinant vaccine construct were characterized and revealed the potency of the vaccine candidate. Then its secondary and tertiary structures, mRNA structure, molecular docking, and immune simulation were predicted using bioinformatics tools. Next, the designed recombinant vaccine construct was synthesized, and cloned into the pET28a vector and expressed in E. coli BL21. Besides, the circular dichroism spectroscopy was utilized for the investigation of the secondary structure changes of the recombinant vaccine construct. The results of the verification assessment of the recombinant vaccine construct expression indicated that in silico analysis was relatively accurate, and relatively change occurred on the protein secondary structure. In our future plan, the vaccine candidate that was confirmed by in silico tools should be validated by further in vitro and in vivo experimental studies. [ABSTRACT FROM AUTHOR]

• Various methods and nanomaterials used to deliver the CRISPR/Cas9 discussed. • Recent progress in the delivery of CRISPR/Cas9 system via nanomaterials presented. • Three different forms of CRISPR/Cas9 discussed in more detail point by point. • Both biological aspect and the structure of nanomaterials discussed. • Outline future challenges in the field of CRISPR/Cas9 components delivery summarized. Exploiting bacterial DNA-acting enzymes and expanding their repertoire for genome engineering have been major technological and conceptual advances in molecular biology over the last decade. The CRISPR-Cas9 system offers many attractive superiorities, such as multiplexing, high precision, low cost, and simplicity compared to other strategies/systems/approaches known to date for gene editing. The efficient co-delivery of Cas9 and single guide RNA(s) into a desired cell and subsequent correct targeting of selected genomic fragment(s) are among the most critical and determining issues for CRISPR-Cas9-based genome engineering. CRISPR/Cas9 components can be transported into target cells via various delivery methods, including physical methods (such as electroporation and microinjection) as well as viral and non-viral methods. Physical and viral methods, with all their privileges, still suffer from disadvantages including induction of immune responses, cell damage, lack of high specificity, etc. We are witnessing a remarkable increase in the employment of nanomaterials as non-viral carriers for the delivery of the CRISPR/Cas9 system. Nanoparticles have so far presented numerous advantages such as ease of synthesis, high efficiency, low cost, size tunability, non-mutagenicity, non-immunogenicity, etc. with regard to the delivery of CRISPR/Cas9. Here, we will review the recent progress in the delivery of CRISPR/Cas9 system components via nanomaterials and outline future challenges. [ABSTRACT FROM AUTHOR]

For the purpose of the current study, hairy root induction in S. bornmuelleri , which is an important medicinal plant, was examined using a particular protocol. Accordingly, some factors such as four strain types of Agrobacterium rhizogenes (A4, A13, MSU440 and ATCC15834), three different explants, namely stem, petiole and leaf, two co-cultivation media, i.e. full and half-MS were studied. Besides, two inoculation methods including injection and immersion as well as three inoculation times (5, 7 and 10 min) were closely taken into account. Utilizing injection method by MSU440 strain, hairy root induction took place in stem explants, and a remarkable increase in transformation frequency (100%) was observed in half-strength MS medium. Methyl jasmonate (MeJA, 100 μM), methyl-b-cyclodextrin (b-CD, 0.7, 7 and 14 mM) and Chitosan (Chi, 50, 100 and 200 mg/l) were used either individually or in a combined way to elicitation. Based on the HPLC results, production of chrysin, wogonin and baicalein increased 9.15, 10.56 and 13.25 times after elicitation of hairy roots by MeJA + Chi. In addition, transcripts of FNSП2 and MYB 7, two important genes involved in the flavonoid biosynthesis pathway, were studies. By applying Chi and MeJA + Chi elicitor, the expression of both genes increased noticeably. It can be concluded that the mentioned hairy root culture system of S. bornmuelleri can be an alternative to flavonoids production. Moreover, there is a direct and positive relationship between the expression of FNSП2 and MYB 7 genes as well as the level of three flavonoids. • Highest transformation frequency of S. bornmuelleri stem explants using MSU440 strain. • Increased expression of FNSП2 and MYB7 genes under elicitor treatments. • MeJA+ Chitosan as the most effective elicitor treatment for increasing the root specific flavonoids. • MYB7 Transcription Factor enhances the gene expression of FNSП2 under elicitor treatments. [ABSTRACT FROM AUTHOR]

Insulin-like growth factor-1 receptor (IGF-1R) is expressed in malignant and normal breast tissue, and its intermittent activation by multiple IGF-1 signaling pathways leads to neoplasm cell proliferation, impaired apoptosis, increased survival, and resistance to cytotoxic therapeutic agents. Therefore, simultaneous suppression of the receptor and its cognate ligand would be a powerful promising strategy inhibiting malignant phenotypes of breast cancer cells. In the present study, Methoxypoly(ethylene glycol) - Poly(caprolactone) was hybridized with Dimethyldioctadecylammonium bromide (DDAB) cationic lipid (mPEG-PCL-DDAB) nanoparticles (NPs) and used as a carrier for simultaneous delivery of lycopene and insulin-like growth factor 1 receptor-specific lycopene encapsulated-mPEG-PCL-DDAB nanoparticle/siRNA to MCF-7 breast cancer cells. Then, the antitumor effects of this construct were evaluated in vitro. The results demonstrated that the synthesized mPEG-PCL-DDAB nanoparticle had suitable physicochemical properties. The use of mPEG-PCL-DDAB nanoparticle-loaded anti-insulin-like growth factor 1 receptor-siRNA and lycopene dramatically induced the process of apoptosis and arrested cell cycle in the MCF-7 tumor cell lines. In general, the findings of this study demonstrated the potency of mPEG-PCL-DDAB nanoparticles for dual delivery of siRNA, and lycopene in breast cancer cell lines followed the induction of apoptosis. [ABSTRACT FROM AUTHOR]

In the present study, we successfully synthesized nanocarriers (NCs) based on Y-shaped miktoarm copolymers, Poly Ethylene Glycol-Lysine-(Poly Caprolactone) 2 (PEG-Lys-PCL 2), which were loaded by baicalein (B) through the nanoprecipitation process to assess their in-vitro and in-vivo properties. We applied various methods and measurements including proton nuclear magnetic resonance (HNMR), dynamic light scattering (DLS), differential scanning calorimetry (DSC), Fourier-transform infrared spectroscopy (FTIR), MTT assay, hemolysis test, lethal dose, real-time PCR, and Morris water maze. The results of DLS indicated that the size and zeta potential of the obtained NCs and B-loaded NCs were acceptable. Also, in-vivo and in-vitro biocompatibility examinations proved that miktoarm-based NCs were safe, and all rats treated with miktoarm-based NCs did not exhibit any remarkable weight loss during the experiment. The results of the Morris water maze (in-vivo test) revealed that the normal saline-treated group, as well as B-miktoarm + Scopolamine (M + B + S) and B-miktoarm-Tween80 + Scopolamine (M + B + T + S) pretreatment groups, spent more time in the target quadrant. Thus, this experiment showed that pretreatment of rats with M + B + S and M + B + T + S had the most effects on spatial memory. According to quantitative PCR analysis, we hypothesized that, in comparison with other experimental groups, pretreatment of rats with M + B + T + S could be more effective in preventing cholinergic dysfunction, brain oxidative stress and cognitive deficits which cause by Scopolamine HBr. This outcome may be partially due to the upregulation of DHCR24, SELADIN, and SIRT6 in entire of the hippocampal region of normal saline-treated and M + B + T + S pretreatment groups. These results may be because mimicking the cell membrane structure would be an excellent feature for miktoarm, and partial coating of Tween-80 can play a critical role for PEG-Lys-PCL 2 -based NCs in crossing the brain cell membrane, and they can easily be uptaken by the cells. Eventually, all of the obtained data confirmed that PEG-Lys-PCL 2 miktoarm star copolymers are suitable for delivering therapeutic agents to the brain for the treatment of Alzheimer's disease (AD). Also, it seems that baicalein should be taken into account as a potent compound for the treatment of AD. [ABSTRACT FROM AUTHOR]

This work described a folic acid conjugated delivery of chrysin-loaded bovine serum albumin nanoparticles, which could overcome the nonspecific targeting disadvantage. Chrysin (5, 7-dihydroxyflavone) is a natural flavonoid which have some significant biological effects on the processes of chemical defense. Chrysin loaded bovine serum albumin nanoparticles (Chrysin-BSA NPs) were synthesized by a simple desolvation procedure. Afterward, folic acid (FA) was conjugated to the surface of Chrysin-BSA NPs by carbodiimide chemistry (Chrysin-BSA-FA NPs). The resultant Chrysin-BSA-FA NPs showed a spherical shape, with a hydrodynamic diameter of 97.5 ± 5.8 nm (mean ± SD) nm and a ζ-potential of −11.3 mV. The in vitro drug release study of chrysin presented a sustained and controlled release pattern. Hemolysis assay and cytotoxicity study results on HFF-2 cell line show that as prepared BSA NPs are biocompatible. Both the Chrysin-BSA NPs and Chrysin-BSA-FA NPs prompted an enhanced cancer cell cytotoxic effect in contrast to chrysin solution. These data recommended that the folate-modified chrysin -loaded vehicle, which demonstrated better biocompatibility and potential superiority, could be a suitable cancer therapy in targeting tumors in the future. [ABSTRACT FROM AUTHOR]

This study described a curcumin (CUR) loaded bovine serum albumin nanoparticles (BSA@CUR NPs), which could solubilize the poorly water-soluble drug and increase the therapeutic efficacy of the drug. BSA@CUR NPs were synthesized by a simple coacervation procedure. The resultant BSA@CUR NPs showed a spherical shape, with a diameter of 92.59 ± 16.75 nm (mean ± SD) nm and a ζ-potential of – 9.19 mV. The in vitro drug release study of CUR showed a sustained and controlled release pattern. Cellular toxicity of BSA NPs was also investigated on HFF2 cell lines. Additionally, a hemolysis test of as prepared NPs were performed for investigation of hemocompatibility. Hemolysis assay and cytotoxicity study results on HFF-2 cell line show that as prepared BSA NPs are biocompatible. The in vitro anticancer activity of the BSA@CUR NPs were performed by MTT assay on MCF-7 cancer cells. These results suggest that BSA@CUR NPs are a new drug delivery system for cancer therapy. [ABSTRACT FROM AUTHOR]

Amphiphilic FA-L-PEG-PCL (PEG: Poly ethylene glycol-hydrophilic segment, FA: Folic acid-targeting agent, L: Lysine-linker, PCL: Poly caprolactone-hydrophobic segment) copolymer was synthesized. Proton nuclear magnetic resonance (HNMR), Fourier-transform infrared spectroscopy (FT-IR), Dynamic light scattering (DLS), atomic force microscopy (AFM), dynamic scanning colorimetry (DSC) were used for characterization of synthesizes copolymers. For determining cytotoxicity of our copolymers, we used from MTT assay, Hemolysis assay and lethal dose 50 (LD50) test. These tests revealed that copolymers had least in vitro and in vivo cytotoxicity and they are categorized as practically none toxic. These copolymers were self-assembled into Round-shaped folate-functionalized micelles in aqueous medium for the folate receptor (FR)-mediated targeted delivery of Tamoxifen (TMX)-the anticancer drug- to cancer cells. The drug loading capacity and in vitro pH responsive controlled release performance showed that these micelles had potential as drug delivery systems (DDS) for hydrophobic anti-cancer drugs such as TMX. FA-L-PEG-PCL micelles was non-cytotoxic in high concentrations. Loaded-TMX micelles obviously showed an increase in killing of the cancer cells. [ABSTRACT FROM AUTHOR]

In this study, iron oxide magnetic bovine serum albumin core–shell nanoparticles (BSA coated IONPs) with narrow particle size distribution were synthesized under one-pot reaction via the desolvation and chemical co-precipitation method. Functionalized IONPs were characterized by X-ray diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM) techniques. Furthermore, vibrating sample magnetometer (VSM) analysis show these nanoparticles (NPs) have an excellent magnetic properties. Cellular toxicity of IONPs was also investigated on HFF2 cell lines. Additionally, a hemolysis test of as prepared core-shell NPs were performed. The presence of albumin as a biomolecule coating on the surface of IONPs showed an improving effect to reduce the cytotoxicity. The properties of the designed NPs propose the BSA coated IONPs as a promising candidate for multifunctional biomedical applications. [ABSTRACT FROM AUTHOR]

Curcumin (CUR) has been associated with anti-inflammatory, antimicrobial, antioxidant, anti-amyloid, and antitumor effects, but its application is limited because of its low aqueous solubility and poor oral bioavailability. To progress the bioavailability and water solubility of CUR, we synthesized five series of mono methoxy poly (ethylene glycol)-poly (ε-caprolactone) (mPEG-PCL) diblock copolymers. The structure of the copolymers was characterized by H NMR, FTIR, DSC and GPC techniques. In this study, CUR was encapsulated within micelles through a single-step nano-precipitation method, leading to formation of CUR-loaded mPEG-PCL (CUR/mPEG-PCL) micelles. The resulting micelles were characterized further by various techniques such as dynamic light scattering (DLS) and atomic force microscopy (AFM). The cytotoxicity of void CUR, mPEG-PCL and CUR/mPEG-PCL micelles was compared to each other by performing MTT assay of the treated MCF-7 and 4T1 cell line. Study of the in vivo pharmacokinetics of the CUR-loaded micelles was also carried out on selected copolymers in comparison with CUR solution formulations. The results showed that the zeta potential of CUR-loaded micelles was about −11.5 mV and the average size was 81.0 nm. CUR was encapsulated into mPEG-PCL micelles with loading capacity of 20.65 ± 0.015% and entrapment efficiency of 89.32 ± 0.34%. The plasma AUC (0–t), t 1/2 and C max of CUR micelles were increased by 52.8, 4.63 and 7.51-fold compared to the CUR solution, respectively. In vivo results showed that multiple injections of CUR-loaded micelles could prolong the circulation time and increase the therapeutic efficacy of CUR. These results suggested that mPEG-PCL micelles would be a potential carrier for CUR. [ABSTRACT FROM AUTHOR]

The application of nanoparticles (NPs) as radio-sensitizers and carriers has opened up a new horizon to overcome the limitations of chemo and radiotherapy. In this study, bovine serum albumin-coated Bi 2 S 3 NPs (Bi 2 S 3 @BSA NPs) were synthesized and evaluated in terms of their ability to be used as a radio-sensitizer and carrier for methotrexate (MTX). Physicochemical properties of MTX conjugated Bi 2 S 3 @BSA NPs (Bi 2 S 3 @BSA-MTX NPs) were characterized by DLS, TEM, FTIR, UV/Vis, and XRD analyses. After the evaluation of cellular uptake and intracellular localization, the cytotoxicity of the combination of Bi 2 S 3 @BSA-MTX NPs and X-Ray radiation was analyzed against the SW480 cell line. The synthesized NPs exhibited spherical-like shapes and homogenous morphology, possessing a hydrodynamic diameter of 140.2 ± 5.71 nm (mean ± SD) and zeta potential of −25 mV. Also, the release study showed that the release of MTX is faster and higher in the presence of the proteinase K enzyme than the absence of the enzyme. The results of in-vitro chemo-radiation therapy indicated that the viability of treated cells with Bi 2 S 3 @BSA-MTX NPs is significantly lower than the cells treated with Bi 2 S 3 @BSA NPs. Furthermore, cells treated with Bi 2 S 3 @BSA-MTX NPs showed a lower degree of viability when combined with X-Ray radiation in comparison with the absence of irradiation, which confirmed the ability of the Bi 2 S 3 @BSA-MTX NPs as radio-sensitizer. [ABSTRACT FROM AUTHOR]