Number of found documents: 307
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Surface functionalization of the biological gold nanoparticles for micro-rna targeting
Pourali, Parastoo; Benada, Oldřich; Benson, Veronika
2021 - English
Among non-viral gene carriers with low toxicity and high transfection efficiency, the use of gold nanoparticles (AuNPs) is of particular interest due to their biocompatibility and special properties. This is the first time we attempted to functionalize the surface of the biological AuNPs in order to conjugate them with antimiR-135b through electrostatic interactions and knockdown the microRNA-135b gene expression inside the cells. A fungal strain, Fusarium oxysporum, was cultured in Sabouraud Dextrose Broth (SDB), centrifuged, and the mycelium-free supernatant was challenged with 1 mmol final concentration of HAuCl4.3H2O and incubated for 24 h at 37°C in a shake flask. AuNPs were characterized by visible spectrophotometry, Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), Energy-Dispersive X-ray spectroscopy (EDS), and a zetasizer. The washed and sterilized AuNPs were used for cytotoxicity and conjugation assays. First transferrin (Tf) and then polyethylenimine (PEI) were used to functionalize and change the surface charge of the AuNPs and then antimiR-135b was conjugated to the AuNPs trough electrostatic interactions. Their association was confirmed by visible spectrophotometry and electrophoresis. Confocal microscopy was used to investigate the internalization of the AuNPs-antimiR-135b complex. The results proved the formation of AuNPs with a maximum absorption peak at 528 nm, round and oval shapes (15-20 nm), and average zeta potential of -21.02 mV. The AuNPs-antimiR-135b showed delayed electrophoresis unlike antimiR-135b or AuNPs alone. Functionalized AuNPs did not cause any toxicity in cell culture and confocal microscopy showed successful transfection of AuNPs-antimiR-135b into the vast majority of 4T1 cells. We concluded that the biological AuNPs were non-toxic and they could carry antimiR-135b to enable gene silencing Keywords: Gold nanoparticles (AuNPs); Biological method; Conjugation; AntimiR-135b Fulltext is available at external website.
Surface functionalization of the biological gold nanoparticles for micro-rna targeting

Among non-viral gene carriers with low toxicity and high transfection efficiency, the use of gold nanoparticles (AuNPs) is of particular interest due to their biocompatibility and special properties. ...

Pourali, Parastoo; Benada, Oldřich; Benson, Veronika
Mikrobiologický ústav, 2021

Enzymatic detoxification of cyanide using cyanide hydratases
Sedova, Anastasia; Rucká, Lenka; Glozlová, Michaela; Novotný, Petr; Martínková, Ludmila; Bojarová, Pavla
2021 - English
Cyanide, a strong chelator of metals in vital enzymes and proteins, is very toxic for most living organisms. Wastewaters from the mining, metallurgical and chemical industries contain significant concentrations of free cyanide. Though it can be largely eliminated by physicochemical methods, these processes may in turn lead to the formation of other waste. In addition, no effective methods have been found to neutralize cyanide spills coming from industrial accidents. The use of enzymes to remove cyanide is an attractive alternative, which is environmentally friendly and applicable to high cyanide concentrations. Cyanide hydratases (CynHs) are of considerable interest for the decomposition of hazardous cyanide wastes. Here we investigate the biochemical properties of new CynHs from Stereum hirsutum and Exidia glandulosa, it is of fundamental importance to preserve the enzyme activity at alkaline pH as cyanide waste is stored under these conditions. Keywords: free cyanide; cyanide hydratase; cyanide spills; cyanide waste Available at various institutes of the ASCR
Enzymatic detoxification of cyanide using cyanide hydratases

Cyanide, a strong chelator of metals in vital enzymes and proteins, is very toxic for most living organisms. Wastewaters from the mining, metallurgical and chemical industries contain significant ...

Sedova, Anastasia; Rucká, Lenka; Glozlová, Michaela; Novotný, Petr; Martínková, Ludmila; Bojarová, Pavla
Mikrobiologický ústav, 2021

Purification of water soluble bioactive polysaccharides from biomass of heterotrophic mutant alga Chlorella vulgaris G11.
Sushytskyi, L.; Lukáč, Pavol; Bleha, R.; Čopíková, J.; Vannucci, Luca; Sinica, A.; Kaštánek, P.
2019 - English
Water extract obtained from biomass of mutant alga Chlorella vulgaris G11 was deproteinized and then purified with AEC on DEAE Sepharose FF and SEC on Sepaphadex G-75 superfine columns. Two high MW (>1000 kDa and 98,3 kDa) polysaccharides (CPS1 and CPS3) were eluted with water and 0.5M NaCl on DEAE column, and heteropolysaccharide with MW of 15 kDa (CPS2) was also eluted with water on DEAE. The former fractions were identified as branched rhamnans, the latter one as branched xylogalactofuranan of unusual structure. CPS1 shown properties of unstable polimer: according to GPC analysis and by using light scattering and RI detectors it was found, that some fraction of polymers (260-800 kDa) have self-assembling properties in aqueous solution, and MW of this fraction decreased during some time while polimer with initially bigger MW (>1000 kDa) became bigger. CPS3 was eluted with 0.3-0.5 M of NaCl on DEAE Sepharose FF column and have MW of 98,3 kDa. Cell toxicity of the products were tested in vitro on cell cultures for different concentrations Keywords: Mutant alga; Chlorella vulgaris G11; chromatography; polymer; cell cultures Available at various institutes of the ASCR
Purification of water soluble bioactive polysaccharides from biomass of heterotrophic mutant alga Chlorella vulgaris G11.

Water extract obtained from biomass of mutant alga Chlorella vulgaris G11 was deproteinized and then purified with AEC on DEAE Sepharose FF and SEC on Sepaphadex G-75 superfine columns. Two high MW ...

Sushytskyi, L.; Lukáč, Pavol; Bleha, R.; Čopíková, J.; Vannucci, Luca; Sinica, A.; Kaštánek, P.
Mikrobiologický ústav, 2019

Nanofiber dressing consisting of antisense rna-functionalized nanodiamonds for therapy of non-healing wounds in diabetic individuals
Neuhoferová, Eva; Petráková, V.; Vocetková, K.; Kindermann, Marek; Křivohlavá, Romana; Benson, Veronika
2018 - English
Non-healing wounds are serious complication in diabetic patients and represent an attractive challenge for development of suitable carrier system possessing constant and localized release of therapeutic biomolecule into the wound without any undesired side effects. Given the fact that these non-healing wounds are result of impaired balance in metalloproteinases synthesized by immune cells residing the wounds, gene therapy offering knock down of such enzymes is of great interest. \nHere we challenged a development of functional and biocompatible wound dressing enabling controlled release of trackable carrier loaded with therapeutic siRNA. Our dressing consists of scaffold from degradable polymer nanofibers enriched with fluorescent nanodiamond particles (FND). We have previously shown the nanodiamond particles are great carriers for antisense RNAs. Their advantages represent high biocompatibility, stable luminescence giving us the possibility to track the carrier system in the wound, and effective release of antisense RNA in the wound. Embedding of nanodiamond-siRNA systems into nanofiber scaffold enables continuous release of siRNA and maintaining the stable siRNA concentration in the wound site resulting in a promotion of wound healing. \nWe developed FND-siRNA complexes specific to MMP-9 that efficiently inhibit the expression of target MMP-9 mRNA. The complexes were embedded into core/shell nanofibers from PVA and PCL, visualized by confocal microscopy, and characterized by electron microscopy. Real-time PCR was used to assess the silencing effect of siRNA that has been delivered to target murine fibroblasts by FND released from nanofiber dressing. Nanofiber system with embedded FNDs was applied on wounds in diabetic animal models to evaluate its suitability regarding short and long term toxicity, efficacy, and handling in vivo. \n Keywords: Electrospun nanofibers; fluorescent nanodiamonds; RNA interference Available at various institutes of the ASCR
Nanofiber dressing consisting of antisense rna-functionalized nanodiamonds for therapy of non-healing wounds in diabetic individuals

Non-healing wounds are serious complication in diabetic patients and represent an attractive challenge for development of suitable carrier system possessing constant and localized release of ...

Neuhoferová, Eva; Petráková, V.; Vocetková, K.; Kindermann, Marek; Křivohlavá, Romana; Benson, Veronika
Mikrobiologický ústav, 2018

BETA-GLUCANS: WHAT APPLICATIONS? A GENERAL OVERVIEW
Vannucci, Luca; Mishra, Rajbardhan; Lukáč, Pavol; Tenti, Paolo; Rajsiglová, Lenka; Větvička, Václav; Šíma, Petr
2018 - English
Glucans are polysaccharides derived from various sources (e.g. mushrooms, algae, yeast, barley). They can target immune receptors, like Dectin-1 expressed on dendritic cells - antigen presenting cells. Otherwise, interest is rising for their use not only for modulation of immunity, but also of cholesterol levels, or as possible vaccine adjuvants, nanocarriers and functional food components. We will shortly review some aspects in human and veterinarian applications. Keywords: Antitumor immune-responses; growth-performance; gastrointestinal-tract Available at various institutes of the ASCR
BETA-GLUCANS: WHAT APPLICATIONS? A GENERAL OVERVIEW

Glucans are polysaccharides derived from various sources (e.g. mushrooms, algae, yeast, barley). They can target immune receptors, like Dectin-1 expressed on dendritic cells - antigen presenting ...

Vannucci, Luca; Mishra, Rajbardhan; Lukáč, Pavol; Tenti, Paolo; Rajsiglová, Lenka; Větvička, Václav; Šíma, Petr
Mikrobiologický ústav, 2018

BETA-GLUCANS: WHAT APPLICATIONS? A GENERAL OVERVIEW
Vannucci, Luca; Mishra, Rajbardhan; Lukáč, Pavol; Tenti, Paolo; Rajsiglová, Lenka; Větvička, Václav; Šíma, Petr
2018 - English
Glucans are polysaccharides derived from various sources (e.g. mushrooms, algae, yeast, barley). They can target immune receptors, like Dectin-1 expressed on dendritic cells - antigen presenting cells. Otherwise, interest is rising for their use not only for modulation of immunity, but also of cholesterol levels, or as possible vaccine adjuvants, nanocarriers and functional food components. We will shortly review some aspects in human and veterinarian applications. Keywords: antitumor immune-responses; growth-performance; gastrointestinal-tract Available at various institutes of the ASCR
BETA-GLUCANS: WHAT APPLICATIONS? A GENERAL OVERVIEW

Glucans are polysaccharides derived from various sources (e.g. mushrooms, algae, yeast, barley). They can target immune receptors, like Dectin-1 expressed on dendritic cells - antigen presenting ...

Vannucci, Luca; Mishra, Rajbardhan; Lukáč, Pavol; Tenti, Paolo; Rajsiglová, Lenka; Větvička, Václav; Šíma, Petr
Mikrobiologický ústav, 2018

Beta-Glucans: What Applications? A General Overview
Vannucci, Luca; Mishra, Rajbardhan; Lukáč, Pavol; Tenti, Paolo; Rajsiglová, Lenka; Větvička, V.; Šíma, Petr
2018 - English
Glucans are polysaccharides derived from various sources (e.g. mushrooms, algae, yeast, barley). They can target immune receptors, like Dectin-1 expressed on dendritic cells - antigen presenting cells. Otherwise, interest is rising for their use not only for modulation of immunity, but also of cholesterol levels, or as possible vaccine adjuvants, nanocarriers and functional food components. We will shortly review some aspects in human and veterinarian applications. Keywords: antitumor immune-responses; growth-performance; gastrointestinal-tract Available at various institutes of the ASCR
Beta-Glucans: What Applications? A General Overview

Glucans are polysaccharides derived from various sources (e.g. mushrooms, algae, yeast, barley). They can target immune receptors, like Dectin-1 expressed on dendritic cells - antigen presenting ...

Vannucci, Luca; Mishra, Rajbardhan; Lukáč, Pavol; Tenti, Paolo; Rajsiglová, Lenka; Větvička, V.; Šíma, Petr
Mikrobiologický ústav, 2018

Nanodiamonds as an innovative system for intracellular delivery of mirna-34a inprostatic cancer therapy
Bitti, G.; Abate, M.; Neuhoferová, Eva; Kindermann, Marek; Petráková, V.; Boccellino, M.; Quagliuolo, L.; Filová, Eva; Benson, Veronika; Caraglia, M.; Amler, Evžen
2018 - English
The microRNA(miRNA)-34a is an important regulator of tumor suppression. It controls the expression of several target proteins involved in cell cycle, differentiation and apoptosis, and antagonizes processes that are necessary for basic cancer cell viability as well as cancer stemness, metastasis, and chemoresistance. It is downregulated in numerous cancer types, including prostatic cancer, and inhibits malignant growth by repressing genes involved in various oncogenic signaling pathways. Given the anti-oncogenic activity of miR-34a, here we proved the substantial benefits of a new therapeutic concept based on nanotechnology delivery of miRNA mimics. In order to monitor the miRNA-34a replacement, we used a fluorescent nanodiamond particles (FND) system with linked miRNA-34a mimic, which was delivered to PC3 and DU145 prostatic cancer cell lines. We used functionalized nanodiamonds coated with polyethylenimine to transfer miRNA-34a into PC3 and DU145 prostatic cancer cell lines and we measured the zeta-potential of these complexes before using them for in vitro experiments. A replacement of miRNA-34 was observed by monitoring levels of miRNA-34 via real-time PCR. Moreover, our in vitro experiments demonstrated that miRNA-34a replacement, using this FND delivery system, decreased viability and induced apoptosis in prostatic cancer cell lines. Our findings suggest the replacement of oncosuppressor miRNA-34a provides an effective strategy for cancer therapy and the FND-based delivery systems seems to be an excellent strategy for a safe and effective targeting of the tumor. Keywords: miR-34a; nanodiamonds; prostate cancer Available at various institutes of the ASCR
Nanodiamonds as an innovative system for intracellular delivery of mirna-34a inprostatic cancer therapy

The microRNA(miRNA)-34a is an important regulator of tumor suppression. It controls the expression of several target proteins involved in cell cycle, differentiation and apoptosis, and antagonizes ...

Bitti, G.; Abate, M.; Neuhoferová, Eva; Kindermann, Marek; Petráková, V.; Boccellino, M.; Quagliuolo, L.; Filová, Eva; Benson, Veronika; Caraglia, M.; Amler, Evžen
Mikrobiologický ústav, 2018

Effect of nanodiamond particles on neuronal tissue
Šimková, Veronika; Benson, Veronika
2018 - English
High-Pressure High-Temperature nanodiamonds are considered to be highly biocompatible due to\ntheir low acute toxicity, which makes them suitable for biomedical applications. However, next to interaction with the cell membrane, nanoscale allows particles to penetrate the plasma membrane and even accumulate in the cell cytoplasm. Recently it brought up questions regarding sustainable nanoparticle application in vivo and potential toxicity hazards. The neuronal tissue consists of neurons and glial cells, and it is separated by a bloodbrain barrier from the peripheral blood system. Neurons are nerve cells forming a complex network of synapses that carry signals in the brain. The neurons are nourished and protected by glial cells. This project aims to find out whether nanodiamonds pass the blood-brain barrier when applied intravenously as a drug carrier and whether they directly interact with the neuronal tissue. Keywords: nanodiamonds; cell cytoplasm; neuronal tissue Available at various institutes of the ASCR
Effect of nanodiamond particles on neuronal tissue

High-Pressure High-Temperature nanodiamonds are considered to be highly biocompatible due to\ntheir low acute toxicity, which makes them suitable for biomedical applications. However, next to ...

Šimková, Veronika; Benson, Veronika
Mikrobiologický ústav, 2018

Development and optimization of the production of recombinant epimerases for pharmaceutical industry. Scale-up of a submerged cultivation technology and down-stream processing
Prell, Aleš; Kujan, Petr; Sobotka, Miroslav; Šafář, Hynek; Vavruška, Tomáš
2017 - English
Development and optimization of the production of recombinant epimerases for pharmaceutical industry. Scale-up of a submerged cultivation technology and down-stream processing. Keywords: enzyme production Available at various institutes of the ASCR
Development and optimization of the production of recombinant epimerases for pharmaceutical industry. Scale-up of a submerged cultivation technology and down-stream processing

Development and optimization of the production of recombinant epimerases for pharmaceutical industry. Scale-up of a submerged cultivation technology and down-stream processing.

Prell, Aleš; Kujan, Petr; Sobotka, Miroslav; Šafář, Hynek; Vavruška, Tomáš
Mikrobiologický ústav, 2017

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