RESUMO
Biotinylated peptide amphiphile (Biotin-PA) nanofibers, are designed as a noncovalent binding location for antigens, which are adjuvants to enhance, accelerate, and prolong the immune response triggered by antigens. Presenting antigens on synthetic Biotin-PA nanofibers generated a higher immune response than the free antigens delivered with a cytosine-phosphate-guanine oligodeoxynucleotides (CpG ODN) (TLR9 agonist) adjuvant. Antigen attached Biotin-PA nanofibers trigger splenocytes to produce high levels of cytokines (IFN-γ, IL-12, TNF-α, and IL-6) and to exhibit a superior cross-presentation of the antigen. Both Biotin-PA nanofibers and CpG ODN induce a Th-1-biased IgG subclass response; however, delivering the antigen with Biotin-PA nanofibers induce significantly greater production of total IgG and subclasses of IgG compared to delivering the antigen with CpG ODN. Contrary to CpG ODN, Biotin-PA nanofibers also enhance antigen-specific splenocyte proliferation and increase the proportion of the antigen-specific CD8(+) T cells. Given their biodegradability and biocompatibility, Biotin-PA nanofibers have a significant potential in immunoengineering applications as a biomaterial for the delivery of a diverse set of antigens derived from intracellular pathogens, emerging viral diseases such as COVID-19, or cancer cells to induce humoral and cellular immune responses against the antigens.
Assuntos
Adjuvantes Imunológicos/química , Nanofibras/química , Peptídeos/química , Peptídeos/imunologia , Adjuvantes Imunológicos/administração & dosagem , Animais , Apresentação de Antígeno , Células Apresentadoras de Antígenos/citologia , Células Apresentadoras de Antígenos/imunologia , Antígenos/administração & dosagem , Antígenos/química , Materiais Biocompatíveis/química , Biotecnologia , Biotina/análogos & derivados , Citocinas/metabolismo , Desenho de Fármacos , Imunidade Celular , Imunidade Humoral , Técnicas In Vitro , Masculino , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Nanofibras/administração & dosagem , Nanofibras/ultraestrutura , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Peptídeos/administração & dosagem , Engenharia de ProteínasRESUMO
Cyclodextrins (CD) are cyclic oligosaccharides produced from the enzymatic degradation of starch as a white powder form; on the other hand, they can be transformed into ultrathin electrospun fiber form by electrospinning technique. The electrospun cyclodextrin fibers (CD-F) can be quite attractive materials to encapsulate bacteria for bioremediation purposes. For instance, CD-F not only serve as a carrier matrix but also it serves as a feeding source for the encapsulated bacteria. In the present study, we demonstrate a facile approach by encapsulation of bacteria into CD-F matrix for wastewater treatment application. The natural and non-toxic properties of CD-F render a better bacterial viability for fibrous biocomposite. The encapsulated bacteria in CD-F exhibit cell viability for more than 7days at 4°C storage condition. Furthermore, we have tested the bioremediation capability of bacteria/CD-F biocomposite for the treatment of heavy metals (Nickel(II) and Chromium(VI)) and textile dye (Reactive Black 5, RB5). The bacteria/CD-F biocomposite has shown removal efficiency of Ni(II), Cr(VI) and RB5 as 70±0.2%, 58±1.4% and 82±0.8, respectively. As anticipated, the pollutants removal capabilities of the bacteria/CD-F was higher compare to free bacteria since bacteria can use CD as an extra carbon source which promotes their growth rate. This study demonstrates that CD-F are suitable platforms for the encapsulation of bacterial cells to develop novel biocomposites that have bioremediation capabilities for wastewater treatment.
Assuntos
Bactérias/metabolismo , Ciclodextrinas/metabolismo , Metais Pesados/metabolismo , Nanofibras/microbiologia , Naftalenossulfonatos/metabolismo , Águas Residuárias/microbiologia , Bactérias/isolamento & purificação , Biodegradação Ambiental , Ciclodextrinas/química , Técnicas Eletroquímicas , Concentração de Íons de Hidrogênio , Metais Pesados/isolamento & purificação , Microscopia Eletrônica de Varredura , Nanofibras/química , Nanofibras/ultraestrutura , Naftalenossulfonatos/isolamento & purificação , Eliminação de Resíduos Líquidos/métodos , Águas Residuárias/química , Poluentes Químicos da Água/química , Poluentes Químicos da Água/isolamento & purificação , Poluentes Químicos da Água/metabolismoRESUMO
Triclosan is considered as an important contaminant and is widely used in personal care products as an antimicrobial agent. This study demonstrates the biodegradation of triclosan by two freshwater microalgae and the acute toxicity of triclosan and 2,4-dichlorophenol. The effects of culture media and light on biodegradation of triclosan and the changing morphology of microalgae were systematically studied. Geitlerinema sp. and Chlorella sp. degraded 82.10% and 92.83% of 3.99 mg/L of triclosan at 10 days, respectively. The microalgal growth inhibition assay confirmed absence of toxic effects of triclosan on Chlorella sp., even at higher concentration (50 mg/L) after 72 h exposure. HPLC analysis showed that 2,4-dichlorophenol was produced as degradation product of triclosan by Geitlerinema sp. and Chlorella sp. This study proved to be beneficial to understand biodegradation and acute toxicity of triclosan by microalgae in order to provide aquatic environmental protection.
Assuntos
Organismos Aquáticos/efeitos dos fármacos , Praguicidas/toxicidade , Triclosan/toxicidade , Biodegradação Ambiental , Bioensaio/métodos , Chlorella/efeitos dos fármacos , Microalgas/efeitos dos fármacos , Poluentes Químicos da Água/toxicidadeRESUMO
In this study, preparation and application of novel biocomposite materials for textile dye removal which are produced by immobilization of specific bacteria onto electrospun nanofibrous webs are presented. A textile dye remediating bacterial isolate, Clavibacter michiganensis, was selected for bacterial immobilization, a commercial reactive textile dye, Setazol Blue BRF-X, was selected as the target contaminant, and electrospun polycaprolactone (PCL) and polylactic acid (PLA) nanofibrous polymeric webs were selected for bacterial integration. Bacterial adhesion onto nanofibrous webs was monitored by scanning electron microscopy (SEM) imaging and optical density (OD) measurements were performed for the detached bacteria. After achieving sufficient amounts of immobilized bacteria on electrospun nanofibrous webs, equivalent web samples were utilized for testing the dye removal capabilities. Both bacteria/PCL and bacteria/PLA webs have shown efficient remediation of Setazol Blue BRF-X dye within 48 h at each tested concentration (50, 100 and 200 mg/L), and their removal performances were very similar to the free-bacteria cells. The bacteria immobilized webs were then tested for five times of reuse at an initial dye concentration of 100 mg/L, and found as potentially reusable with higher bacterial immobilization and faster dye removal capacities at the end of the test. Overall, these findings suggest that electrospun nanofibrous webs are available platforms for bacterial integration and the bacteria immobilized webs can be used as starting inocula for use in remediation of textile dyes in wastewater systems.
Assuntos
Bactérias/metabolismo , Biodegradação Ambiental , Poluentes Químicos da Água/isolamento & purificação , Corantes/isolamento & purificação , Poliésteres , Indústria Têxtil , Águas ResiduáriasRESUMO
Thymol (THY)/γ-Cyclodextrin(γ-CD) inclusion complex (IC) encapsulated electrospun zein nanofibrous webs (zein-THY/γ-CD-IC-NF) were fabricated as a food packaging material. The formation of THY/γ-CD-IC (1:1 and 2:1) was proved by experimental (X-ray diffraction (XRD), thermal gravimetric analysis (TGA), 1H NMR) and computational techniques. THY/γ-CD-IC (2:1) exhibited higher preservation rate and stability than THY/γ-CD-IC (1:1). It is worth mentioning that zein-THY/γ-CD-IC-NF (2:1) preserved much more THY as observed in TGA and stability of THY/γ-CD-IC (2:1) was higher, as shown by a modelling study. Therefore, much more THY was released from zein-THY/γ-CD-IC-NF (2:1) than zein-THY-NF and zein-THY/γ-CD-IC-NF (1:1). Similarly, antibacterial activity of zein-THY/γ-CD-IC-NF (2:1) was higher than zein-THY-NF and zein-THY/γ-CD-IC-NF (1:1). It was demonstrated that zein-THY/γ-CD-IC-NF (2:1) was most effective in inhibiting the growth of bacteria on meat samples. These webs show potential application as an antibacterial food packaging material.
Assuntos
Nanofibras , Antibacterianos , Ciclodextrinas , Embalagem de Alimentos , Timol , ZeínaRESUMO
Burn injuries are one of the most common types of trauma worldwide, and their unique physiology requires the development of specialized therapeutic materials for their treatment. Here, we report the use of synthetic, functional and biodegradable peptide nanofiber gels for the improved healing of burn wounds to alleviate the progressive loss of tissue function at the post-burn wound site. These bioactive nanofiber gels form scaffolds that recapitulate the structure and function of the native extracellular matrix through signaling peptide epitopes, which can trigger angiogenesis through their affinity to basic growth factors. In this study, the angiogenesis-promoting properties of the bioactive scaffolds were utilized for the treatment of a thermal burn model. Following the excision of necrotic tissue, bioactive gels and control solutions were applied topically onto the wound area. The wound healing process was evaluated at 7, 14 and 21 days following injury through histological observations, immunostaining and marker RNA/protein analysis. Bioactive peptide nanofiber-treated burn wounds formed well-organized and collagen-rich granulation tissue layers, produced a greater density of newly formed blood vessels, and exhibited increased re-epithelialization and skin appendage development with minimal crust formation, while non-bioactive peptide nanofibers and the commercial wound dressing 3M™ Tegaderm™ did not exhibit significant efficiency over sucrose controls. Overall, the heparin-mimetic peptide nanofiber gels increased the rate of repair of burn injuries and can be used as an effective means of facilitating wound healing.
Assuntos
Queimaduras/terapia , Géis/química , Heparina/química , Nanofibras/química , Peptídeos/química , Alicerces Teciduais/química , Animais , Dicroísmo Circular , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Varredura , Cicatrização/fisiologiaRESUMO
The volatility and limited water solubility of linalool is a critical issue to be solved. Here, we demonstrated the electrospinning of polymer-free nanofibrous webs of cyclodextrin/linalool-inclusion complex (CD/linalool-IC-NFs). Three types of modified cyclodextrin (HPßCD, MßCD, and HPγCD) were used to electrospin CD/linalool-IC-NFs. Free-standing CD/linalool-IC-NFs facilitate maximum loading of linalool up to 12% (w/w). A significant amount of linalool (45-89%) was preserved in CD/linalool-IC-NFs, due to enhancement in the thermal stability of linalool by cyclodextrin inclusion complexation. Remarkably, CD/linalool-IC-NFs have shown fast-dissolving characteristics in which these nanofibrous webs dissolved in water within two seconds. Furthermore, linalool release from CD/linalool-IC-NFs inhibited growth of model Gram-negative (E. coli) and Gram-positive (S. aureus) bacteria to a great extent. Briefly, characteristics of liquid linalool have been preserved in a solid nanofiber form and designed CD/linalool-IC-NFs confer high loading capacity, enhanced shelf life and strong antibacterial activity of linalool.
Assuntos
Ciclodextrinas , Monoterpenos , Nanofibras , Monoterpenos Acíclicos , Antibacterianos , Escherichia coli , Staphylococcus aureus , beta-CiclodextrinasRESUMO
In this study, preparation and application of novel biocomposite materials that were produced by encapsulation of bacterial cells within electrospun nanofibrous webs are described. A commercial strain of Pseudomonas aeruginosa which has methylene blue (MB) dye remediation capability was selected for encapsulation, and polyvinyl alcohol (PVA) and polyethylene oxide (PEO) were selected as the polymer matrices for the electrospinning of bacteria encapsulated nanofibrous webs. Encapsulation of bacterial cells was monitored by scanning electron microscopy (SEM) and fluorescence microscopy, and the viability of encapsulated bacteria was checked by live/dead staining and viable cell counting assay. Both bacteria/PVA and bacteria/PEO webs have shown a great potential for remediation of MB, yet bacteria/PEO web has shown higher removal performances than bacteria/PVA web, which was probably due to the differences in the initial viable bacterial cells for those two samples. The bacteria encapsulated electrospun nanofibrous webs were stored at 4°C for three months and they were found as potentially storable for keeping encapsulated bacterial cells alive. Overall, the results suggest that electrospun nanofibrous webs are suitable platforms for preservation of living bacterial cells and they can be used directly as a starting inoculum for bioremediation of water systems.
Assuntos
Bactérias/metabolismo , Biodegradação Ambiental , Azul de Metileno/metabolismo , Nanofibras/microbiologia , Água/metabolismoRESUMO
This study evaluated the effect of intra-amniotic synbiotic inclusion and continued synbiotic supplementation in the diet on the performance, intestinal epithelium integrity, and cecal microflora of broiler chickens. In Experiment 1, 510 eggs containing viable embryos were divided into 3 groups of 170 eggs each. The first group was not injected and served as a negative control ( NC: ). The next group was injected with 0.9% NaCl and was the positive control ( PC: ). The synbiotic-injected group ( S: ) was injected with a 0.5% inulin and 1 × 106 Enterococcus faecium solution. The non-injected and synbiotic injected groups were further divided into 2 sets for Experiment 2 and the birds were offered either a basal or synbiotic supplemented diet (1% inulin and 2 × 109 E. faecium cfu/kg feed). One hundred ninety-six broiler hatchlings were randomly allocated in a 2 × 2 factorial arrangement that included an intra-amniotic treatment (non-injected or synbiotic injected) and a dietary treatment (basal or synbiotic supplemented diet). The results showed that the administration of an intra-amniotic synbiotic to embryonated eggs on d 17 of incubation did not affect the hatchability or hatching weight of the birds. However, intra-amniotic synbiotic inclusion had a positive effect on FCR at d 0 to 42 (P = 0.041) and d 22 to 42 (P = 0.036). There was no significant interaction effect on the growth performance of the birds between the intra-amniotic and dietary synbiotic treatment during different or entire experimental periods. Villus height and goblet and proliferating cell nuclear antigen ( PCNA: ) positive cell counts were positively influenced by intra-amniotic and dietary synbiotic treatments. Our results also indicated that intra-amniotic synbiotic injection followed by dietary supplementation with a synbiotic significantly increased Lactobacillus colonization and decreased coliform population in the broiler cecum. Cecal butyric acid concentration increased proportionally to the cecal Lactobacillus count with dietary synbiotic supplementation on d 42. In summary, combined intra-amniotic and dietary synbiotic treatments improved broiler intestinal integrity and increased cecal beneficial bacteria populations.
Assuntos
Ceco/microbiologia , Galinhas/microbiologia , Galinhas/fisiologia , Dieta/veterinária , Ácidos Graxos Voláteis/metabolismo , Intestinos/citologia , Simbióticos/administração & dosagem , Ração Animal/análise , Animais , Galinhas/anatomia & histologia , Galinhas/crescimento & desenvolvimento , Células Epiteliais/citologia , Injeções/veterinária , Masculino , Carne/análise , Óvulo/química , Distribuição AleatóriaRESUMO
The binding interaction between two dicationic styrylimidazo[1,2-a]pyridinium dyes and human serum albumin (HSA) was investigated at physiological conditions using fluorescence, UV-vis absorption, and circular dichroism (CD) spectroscopies. Analysis of the fluorescence titration data at different temperatures suggested that the fluorescence quenching mechanism of HSA by these dyes was static. The calculated thermodynamic parameters (ΔG°, ΔH° and ΔS°) indicated that hydrogen bonding and van der Waals forces played a major role in the formation of the dye-HSA complex. Binding distances (r) between dyes and HSA were calculated according to Förster's non-radiative energy transfer theory. Studies of conformational changes of HSA using CD measurements indicate that the α-helical content of the protein decreased upon binding of the dyes. Copyright © 2016 John Wiley & Sons, Ltd.
Assuntos
Corantes Fluorescentes/química , Imidazóis/química , Compostos de Piridínio/química , Albumina Sérica/química , Sítios de Ligação , Cátions/química , Dicroísmo Circular , Humanos , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta , TermodinâmicaRESUMO
The design and development of vaccines, which can induce cellular immunity, particularly CD8+ T cells hold great importance since these cells play crucial roles against cancers and viral infections. Covalent conjugation of antigen and adjuvant molecules has been used for successful promotion of immunogenicity in subunit vaccines; however, the stimulation of the CD8+ T-cell responses by this approach has so far been limited. This study demonstrates a modular system based on noncovalent attachment of biotinylated antigen to a hybrid nanofiber system consisting of biotinylated self-assembling peptide and CpG oligodeoxynucleotides (ODN) molecules, via biotin-streptavidin interaction. These peptide/oligonucleotide hybrid nanosystems are capable of bypassing prior limitations related with inactivated or live-attenuated virus vaccines and achieve exceptionally high CD8+ T-cell responses. The nanostructures are found to trigger strong IgG response and effectively modulate cross-presentation of their antigen "cargo" through close proximity between the antigen and peptide/ODN adjuvant system. In addition, the biotinylated peptide nanofiber system is able to enhance antigen uptake and induce the maturation of antigen-presenting cells. Due to its versatility, biocompatibility, and biodegradability with a broad variety of streptavidin-linked antigens, the nanosystem shown here can be utilized as an efficient strategy for new vaccine development.
RESUMO
We have proposed a new strategy for preparing free-standing nanofibrous webs from an inclusion complex (IC) of a well-known flavor/fragrance compound (limonene) with three modified cyclodextrins (HPßCD, MßCD, and HPγCD) via electrospinning (CD/limonene-IC-NFs) without using a polymeric matrix. The experimental and computational modeling studies proved that the stoichiometry of the complexes was 1:1 for CD/limonene systems. MßCD/limonene-IC-NF released much more limonene at 37, 50, and 75 °C than HPßCD/limonene-IC-NF and HPγCD/limonene-IC-NF because of the greater amount of preserved limonene. Moreover, MßCD/limonene-IC-NF has released only 25% (w/w) of its limonene, whereas HPßCD/limonene-IC-NF and HPγCD/limonene-IC-NF released 51 and 88% (w/w) of their limonene in 100 days, respectively. CD/limonene-IC-NFs exhibited high antibacterial activity against E. coli and S. aureus. The water solubility of limonene increased significantly and CD/limonene-IC-NFs were dissolved in water in a few seconds. In brief, CD/limonene-IC-NFs with fast-dissolving character enhanced the thermal stability and prolonged the shelf life along with antibacterial properties could be quite applicable in food and oral care applications.
Assuntos
Antibacterianos/química , Ciclodextrinas/química , Cicloexenos/química , Portadores de Fármacos/química , Nanofibras/química , Terpenos/química , Computadores Moleculares , Cristalografia por Raios X , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Escherichia coli/efeitos dos fármacos , Limoneno , Espectroscopia de Ressonância Magnética , Microscopia Eletrônica de Varredura , Óleos Voláteis/química , Tamanho da Partícula , Solubilidade , Staphylococcus aureus/efeitos dos fármacos , TermogravimetriaRESUMO
Triclosan is an antimicrobial agent and a persistent pollutant. The biodegradation of triclosan is dependent on many variables including the biodegradation organism and the environmental conditions. Here, we evaluated the triclosan degradation potential of two fungi strains, Rhodotorula mucilaginosa and Penicillium sp., and the rate of its turnover to 2,4-dichlorophenol (2,4-DCP). Both of these strains showed less susceptibility to triclosan when grown in minimal salt medium. In order to further evaluate the effects of environmental conditions on triclosan degradation, three different culture conditions including original thermal power plant wastewater, T6 nutrimedia and ammonium mineral salts medium were used. The maximum triclosan degradation yield was 48% for R. mucilaginosa and 82% for Penicillium sp. at 2.7 mg/L triclosan concentration. Biodegradation experiments revealed that Penicillium sp. was more tolerant to triclosan. Scanning electron microscopy micrographs also showed the morphological changes of fungus when cells were treated with triclosan. Overall, these fungi strains could be used as effective microorganisms in active uptake (degradation) and passive uptake (sorption) of triclosan and their efficiency can be increased by optimizing the culture conditions.
Assuntos
Meios de Cultura/análise , Poluentes Ambientais/metabolismo , Penicillium/metabolismo , Rhodotorula/metabolismo , Triclosan/metabolismo , Anti-Infecciosos Locais/metabolismo , Biodegradação Ambiental , Clorofenóis/metabolismo , Águas Residuárias/análiseRESUMO
Phenylboronic acid-functionalized, Ag shell-coated, magnetic, monodisperse polymethacrylate microspheres equipped with a glycoprotein-sensitive sandwich system were proposed as a surface-enhanced Raman scattering (SERS) substrate for quantitative determination of glycated hemoglobin (HbA1c). The magnetization of the SERS tag and the formation of the Ag shell on the magnetic support were achieved using the bifunctional reactivity of newly synthesized polymethacrylate microspheres. The hemolysate of human red blood cells containing both HbA1c and nonglycated hemoglobin was used for determination of HbA1c. The working principle of the proposed SERS tag is based on the immobilization of HbA1c by cyclic boronate ester formation between glycosyl residues of HbA1c and boronic acid groups of magnetic polymethacrylate microspheres and the binding of p-aminothiophenol (PATP)-functionalized Ag nanoparticles (Ag NPs) carrying another boronic acid ligand via cyclic boronate ester formation via unused glycosyl groups of bound HbA1c. Then, in situ formation of a Raman reporter, 4,4'-dimercaptoazobenzene from PATP under 785 nm laser irradiation allowed for the quantification of HbA1c bound onto the magnetic SERS tag, which was proportional to the HbA1c concentration in the hemolysate of human erythrocytes. The sandwich system provided a significant enhancement in the SERS signal intensity due to the plasmon coupling between Ag NPs and Ag shell-coated magnetic microspheres, and low HbA1c concentrations down to 50 ng/mL could be detected. The calibration curve obtained with a high correlation coefficient between the SERS signal intensity and HbA1c level showed the usability of the SERS protocol for the determination of the HbA1c level in any person.
Assuntos
Ácidos Borônicos/química , Eritrócitos/química , Hemoglobinas Glicadas/análise , Hemólise , Análise Espectral Raman , HumanosRESUMO
Oligonucleotides are promising drug candidates due to the exceptionally high specificity they exhibit toward their target DNA and RNA sequences. However, their poor pharmacokinetic and pharmacodynamic properties, in conjunction with problems associated with their internalization by cells, necessitates their delivery through specialized carrier systems for efficient therapy. Here, we investigate the effects of carrier morphology on the cellular internalization mechanisms of oligonucleotides by using self-assembled fibrous or spherical peptide nanostructures. Size and geometry were both found to be important parameters for the oligonucleotide internalization process; direct penetration was determined to be the major mechanism for the internalization of nanosphere carriers, whereas nanofibers were internalized by clathrin- and dynamin-dependent endocytosis pathways. We further showed that glucose conjugation to carrier nanosystems improved cellular internalization in cancer cells due to the enhanced glucose metabolism associated with oncogenesis, and the internalization of the glucose-conjugated peptide/oligonucleotide complexes was found to be dependent on glucose transporters present on the surface of the cell membrane.
Assuntos
Nanofibras , Nanosferas , Clatrina , Endocitose , Humanos , Oligonucleotídeos , PeptídeosRESUMO
Following the rapid uptake of contaminants in the first few hours of exposure, plants typically attempt to cope with the toxic burden by releasing part of the sorbed material back into the environment. The present study investigates the general trends in the release profiles of different metal(loid)s in the aquatic macrophyte Lemna minor and details the correlations that exist between the release of metal(loid) species. Water samples with distinct contamination profiles were taken from Nilüfer River (Bursa, Turkey), Yeniçaga Lake (Bolu, Turkey), and Beysehir Lake (Konya, Turkey) and used for release studies; 36 samples were tested in total. Accumulation and release profiles were monitored over five days for 11 metals and a metalloid ((208)Pb, (111)Cd, (52)Cr,(53)Cr,(60)Ni,(63)Cu,(65)Cu,(75)As,(55)Mn, (137)Ba, (27)Al, (57)Fe, (66)Zn,(68)Zn) and correlation, cluster and principal component analyses were employed to determine the factors that affect the release of these elements. Release profiles of the tested metal(loid)s were largely observed to be distinct; however, strong correlations have been observed between certain metal pairs (Cr/Ni, Cr/Cu, Zn/Ni) and principal component analysis was able to separate the metal(loid)s into three well-resolved groups based on their release.
Assuntos
Araceae/metabolismo , Metais/metabolismo , Poluentes da Água/metabolismo , Biodegradação Ambiental , Água Doce , TurquiaRESUMO
We describe the binding characteristics of two natural borates (colemanite and ulexite) to calf thymus DNA by UV-vis absorbance spectroscopy, circular dichroism (CD) spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, and a competitive DNA binding assay. Our results suggest that colemanite and ulexite interact with calf thymus DNA under a non-intercalative mode of binding and do not alter the secondary structure of the DNA helix. The FT-IR spectroscopy results indicate that the two borates might interact with DNA through sugar-phosphate backbone binding.
Assuntos
Boratos/química , DNA/química , Animais , Bovinos , Dicroísmo Circular , Etídio/química , Conformação de Ácido Nucleico , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier , TimoRESUMO
This study demonstrates the acute toxicity of lanthanum oxide nanoparticles (La2O3 NP) on two sentinel aquatic species, fresh-water microalgae Chlorella sp. and the crustacean Daphnia magna. The morphology, size and charge of the nanoparticles were systematically studied. The algal growth inhibition assay confirmed absence of toxic effects of La2O3 NP on Chlorella sp., even at higher concentration (1000 mg L(-1)) after 72 h exposure. Similarly, no significant toxic effects were observed on D. magna at concentrations of 250 mg L(-1) or less, and considerable toxic effects were noted in higher concentrations (effective concentration [EC50] 500 mg L(-1); lethal dose [LD50] 1000 mg L(-1)). In addition, attachment of La2O3 NP on aquatic species was demonstrated using microscopy analysis. This study proved to be beneficial in understanding acute toxicity in order to provide environmental protection as part of risk assessment strategies.
Assuntos
Lantânio/toxicidade , Nanopartículas/toxicidade , Óxidos/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Clorófitas , Daphnia , Água Doce , Microalgas , Testes de ToxicidadeRESUMO
A drug delivery system designed specifically for oligonucleotide therapeutics can ameliorate the problems associated with the in vivo delivery of these molecules. The internalization of free oligonucleotides is challenging, and cytotoxicity is the main obstacle for current transfection vehicles. To develop nontoxic delivery vehicles for efficient transfection of oligonucleotides, we designed a self-assembling peptide amphiphile (PA) nanosphere delivery system decorated with cell penetrating peptides (CPPs) containing multiple arginine residues (R4 and R8), and a cell surface binding peptide (KRSR), and report the efficiency of this system in delivering G-3129, a Bcl-2 antisense oligonucleotide (AON). PA/AON (peptide amphiphile/antisense oligonucleotide) complexes were characterized with regards to their size and secondary structure, and their cellular internalization efficiencies were evaluated. The effect of the number of arginine residues on the cellular internalization was investigated by both flow cytometry and confocal imaging, and the results revealed that uptake efficiency improved as the number of arginines in the sequence increased. The combined effect of cell penetration and surface binding property on the cellular internalization and its uptake mechanism was also evaluated by mixing R8-PA and KRSR-PA. R8 and R8/KRSR decorated PAs were found to drastically increase the internalization of AONs compared to nonbioactive PA control. Overall, the KRSR-decorated self-assembled PA nanospheres were demonstrated to be noncytotoxic delivery vectors with high transfection rates and may serve as a promising delivery system for AONs.
Assuntos
Peptídeos Penetradores de Células/química , Nanosferas/química , Oligonucleotídeos Antissenso/química , Sobrevivência Celular/efeitos dos fármacos , Peptídeos Penetradores de Células/efeitos adversos , Dicroísmo Circular , Citometria de Fluxo , Humanos , Células MCF-7 , Microscopia Confocal , Microscopia Eletrônica de TransmissãoRESUMO
Lysozyme is an important polypetide used in medical and food applications. We report a novel magnetic strong cation exchange beads for efficient purification of lysozyme from chicken egg white. Magnetic chitosan (MCHT) beads were synthesized via phase inversion method, and then grafted with poly(glycidyl methacrylate) (p(GMA)) via the surface-initiated atom transfer radical polymerization (SI-ATRP). Epoxy groups of the grafted polymer, were modified into strong cation-exchange groups (i.e., sulfonate groups) in the presence of sodium sulfite. The MCTH and MCTH-g-p(GMA)-SO3H beads were characterized by ATR-FTIR, SEM, and VSM. The sulphonate groups content of the modified MCTH-g-p(GMA)-4 beads was found to be 0.53mmolg(-1) of beads by the potentiometric titration method. The MCTH-g-p(GMA)-SO3H beads were first used as an ion-exchange support for adsorption of lysozyme from aqueous solution. The influence of different experimental parameters such as pH, contact time, and temperature on the adsorption process was evaluated. The maximum adsorption capacity was found to be 208.7mgg(-1) beads. Adsorption of lysozyme on the MCTH-g-p(GMA)-SO3H beads fitted to Langmuir isotherm model and followed the pseudo second-order kinetic. More than 93% of the adsorbed lysozyme was desorbed using Na2CO3 solution (pH 11.0). The purity of the lysozyme was checked by HPLC and SDS gel electrophoresis. In addition, the MCTH-g-p(GMA)-SO3H beads prepared in this work showed promising potential for separation of various anionic molecules.