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1.
BackgroundAlthough nanoparticles (NPs) have many advantages, it has been proved that they may be absorbed by and have toxic effects on the human body. Recent research has tried to evaluate and compare the nanotoxicity of gold nanoparticles (AuNPs) produced by two types of microorganisms in vitro by two different methods. AuNPs were produced by Bacillus cereus and Fusarium oxysporum, and their production was confirmed by visible spectral, transmission electron microscope, and X-ray diffraction (XRD) analyses. The human fibroblast cell line CIRC-HLF was treated with AuNPs, and the induced nanotoxicity was measured using direct microscopic and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays.ResultsThe results showed that the produced AuNPs had a maximum absorbance peak around 510–530 nanometer (nm), with spherical, hexagonal, and octagonal shapes and average sizes around 20–50 nm. The XRD results confirmed the presence of GNPs in the microbial culture supernatants. An MTT assay showed that GNPs had dose-dependent toxic effects, and microscopic analysis showed that GNPs induced cell abnormalities in doses lower than the determined half-maximal inhibitory concentrations (IC50s).ConclusionsIn conclusion, the biologically produced AuNPs had toxic effects in the cell culture, and direct techniques such as microscopic evaluation instead of indirect methods such as MTT assay were more useful for assessing the nanotoxicity of the biologically produced AuNPs. Thus, the use of only MTT assay for nanotoxicity evaluation of AuNPs is not desirable.  相似文献   

2.
Wang L  Li PC 《Biomicrofluidics》2010,4(3):32209
Two simple gold nanoparticle (GNP)-based DNA analysis methods using a microfluidic device are presented. In the first method, probe DNA molecules are immobilized on the surface of a self-assembled submonolayer of GNPs. The hybridization efficiency of the target oligonulceotides was improved due to nanoscale spacing between probe molecules. In the second method, target DNA molecules, oligonulceotides or polymerase chain reaction (PCR) amplicons, are first bound to GNPs and then hybridized to the immobilized probe DNA on a glass slide. With the aid of GNPs, we have successfully discriminated, at room temperature, between two PCR amplicons (derived from closely related fungal pathogens, Botrytis cinerea and Botrytis squamosa) with one base-pair difference. DNA analysis on the microfluidic chip avoids the use of large sample volumes, and only a small amount of oligonucelotides (8 fmol) or PCR products (3 ng), was needed in the experiment. The whole procedure was accomplished at room temperature in 1 h, and apparatus for high temperature stringency was not required.  相似文献   

3.
BackgroundCyclodextrin glucanotransferase (CGTase) from Amphibacillus sp. NPST-10 was covalently immobilized onto amino-functionalized magnetic double mesoporous core–shell silica nanospheres (mag@d-SiO2@m-SiO2-NH2), and the properties of the immobilized enzyme were investigated. The synthesis process of the nanospheres included preparing core magnetic magnetite (Fe3O4) nanoparticles, coating the Fe3O4 with a dense silica layer, followed by further coating with functionalized or non-functionalized mesoporous silica shell. The structure of the synthesized nanospheres was characterized using TEM, XRD, and FT-IR analyses. CGTase was immobilized onto the functionalized and non-functionalized nanospheres by covalent attachment and physical adsorption.ResultsThe results indicated that the enzyme immobilization by covalent attachment onto the activated mag@d-SiO2@m-SiO2-NH2, prepared using anionic surfactant, showed highest immobilization yield (98.1%), loading efficiency (96.2%), and loading capacity 58 µg protein [CGTase]/mg [nanoparticles]) which were among the highest yields reported so far for CGTase. Compared with the free enzyme, the immobilized CGTase demonstrated a shift in the optimal temperature from 50°C to 50–55°C, and showed a significant enhancement in the enzyme thermal stability. The optimum pH values for the activity of the free and immobilized CGTase were pH 8 and pH 8.5, respectively, and there was a significant improvement in pH stability of the immobilized enzyme. Moreover, the immobilized CGTase exhibited good operational stability, retaining 56% of the initial activity after reutilizations of ten successive cycles.ConclusionThe enhancement of CGTase properties upon immobilization suggested that the applied nano-structured carriers and immobilization protocol are promising approach for industrial bioprocess for production of cyclodextrins using immobilized CGTase.  相似文献   

4.
The poor solubility of many newly discovered drugs has resulted in numerous challenges for the time-controlled release of therapeutics. In this study, an advanced drug delivery platform to encapsulate and deliver hydrophobic drugs, consisting of poly (lactic-co-glycolic acid) (PLGA) nanoparticles incorporated within poly (ethylene glycol) (PEG) microgels, was developed. PLGA nanoparticles were used as the hydrophobic drug carrier, while the PEG matrix functioned to slow down the drug release. Encapsulation of the hydrophobic agents was characterized by fluorescence detection of the hydrophobic dye Nile Red within the microgels. In addition, the microcomposites prepared via the droplet-based microfluidic technology showed size tunability and a monodisperse size distribution, along with improved release kinetics of the loaded cargo compared with bare PLGA nanoparticles. This composite system has potential as a universal delivery platform for a variety of hydrophobic molecules.  相似文献   

5.
This paper presents a convenient strategy to modify the surface of whole-Teflon microfluidic chips by coating the channel walls with a thin layer of polydopamine (PDA) film, which is formed by oxidation-induced self-polymerization of dopamine in alkaline solution. Two coating strategies, static incubation and dynamic flow, are demonstrated and used for tuning the physical and chemical properties of the coated channel walls. The functionalized surfaces were investigated with the contact angle, X-ray photoelectron spectroscopy, and atomic force microscopy measurements. The coating time was optimized according to the fluorescent intensity of the green fluorescent protein immobilized on the modified surface. Applications of the PDA-modified Teflon microchips in bioanalysis were demonstrated with a typical sandwich immunoassay. Moreover, long-term cell culture experiments on modified and native Teflon chips revealed that the chip biocompatibility can be greatly improved with PDA coating. The results indicate that the surface properties of the Teflon can be easily controlled by the PDA modification, thus greatly expanding the application scope of whole-Teflon chips for various chemical and biological research fields.  相似文献   

6.
A new strategy for magnetically manipulating and isolating adherent cells with extremely high post-collection purity and viability is reported. Micromolded magnetic elements (termed microrafts) were fabricated in an array format and used as culture surfaces and carriers for living, adherent cells. A poly(styrene-co-acrylic acid) polymer containing well dispersed magnetic nanoparticles was developed for creating the microstructures by molding. Nanoparticles of γFe(2)O(3) at concentrations up to 1% wt.∕wt. could be used to fabricate microrafts that were optically transparent, highly magnetic, biocompatible, and minimally fluorescent. To prevent cellular uptake of nanoparticles from the magnetic polymer, a poly(styrene-co-acrylic acid) layer lacking γFe(2)O(3) nanoparticles was placed over the initial magnetic microraft layer to prevent cellular uptake of the γFe(2)O(3) during culture. The microraft surface geometry and physical properties were altered by varying the polymer concentration or layering different polymers during fabrication. Cells plated on the magnetic microrafts were visualized using standard imaging techniques including brightfield, epifluorescence, and confocal microscopy. Magnetic microrafts possessing cells of interest were dislodged from the array and efficiently collected with an external magnet. To demonstrate the feasibility of cell isolation using the magnetic microrafts, a mixed population of wild-type cells and cells stably transfected with a fluorescent protein was plated onto an array. Microrafts possessing single, fluorescent cells were released from the array and magnetically collected. A post-sorting single-cell cloning rate of 92% and a purity of 100% were attained.  相似文献   

7.
Ultrasmall superparamagnetic iron oxide nanoparticles coated with gummic acid have been investigated as possible constituents of aqueous ferrofluids for biomedical applications and especially for MRI contrast agent. The structural characteristics and the size of the nanoparticles have been analyzed as well as the magnetic properties. In order to evaluate any possible capabilities as a contrast agent, the relaxation time, T2, of hydrogen protons in the colloidal solutions of nanoparticles have been measured in order to gain information on the relaxation behavior compared to other MRI contrast agents. The in vitro cytotoxicity of the obtained magnetic nanoparticles of iron oxide coated with gummic acid was investigated by two separate methods (MTT and FACS analysis) and by using three different normal and transformed cell lines. Our results showed that the synthesized nanoparticles had no toxic effect on any of the cell lines used.  相似文献   

8.
Building on recent breakthroughs in the field of microfluidic-based capture of rare cancer cells circulating in the blood, the present article reports on the use of Herceptin functionalized PDMS devices designed to efficiently capture from blood cancer cells, overexpressing the tyrosine kinase human epidermal growth factor receptor (HER2). The identification of patients overexpressing HER2 is critical as it typically associates with an aggressive disease course in breast cancer and poor prognosis. Importantly, HER2 positive patients have been found to significantly benefit from Herceptin (Trastuzumab), a humanized monoclonal antibody (MAb) against HER2. Disposable PDMS devices prepared using standard soft lithography were functionalized by the plasma polymerization of an epoxy-containing monomer. The epoxy-rich thin film (AGEpp) thus created could be conjugated with Herceptin either directly or through a polyethylene glycol interlayer. The properties and reactivity toward the monoclonal antibody conjugation of these coatings were determined using x-ray photoelectron spectroscopy; direct conjugation provided a good compromise in reactivity and resistance to biologically nonspecific fouling and was selected. Using the breast cancer cell line SK-BR-3 as a model for cells overexpressing HER2, the immunocapture efficacy of the Herceptin functionalized PDMS was demonstrated in model studies. Validation studies confirmed the ability of the device to efficiently capture (～80% capture yield) HER2 positive cells from full blood.  相似文献   

9.
胡文祥  恽榴红 《科技通报》1995,11(2):115-119
用热力学辅之以分子力学和量子化学来研究药物与受体相互使用，而其中的轨道作用可用药效基集团中各点轨道作用的线性组合来表达，各点轨道作用可用局域轨道模型来处理，部分概念和公式在药物分子与胆碱能受体相互作用研究中得到了支持和验证。  相似文献   

10.
明胶水溶液中银离子的光还原   总被引：3，自引：0，他引：3  
利用紫外线辐射ＡｇＮＯ３明胶（ＣＨ３）２ＣＨＯＨ 水溶液制备了银纳米颗粒,并用吸收光谱和透射电镜跟踪研究了其形成过程．发现增大明胶和银离子浓度可以加速银颗粒的生成,其粒径随明胶浓度增大而减小,银颗粒的光还原过程属于准一级反应;明胶保护下的银纳米颗粒具有很高的稳定性．提出了ＡｇＮＯ３明胶（ＣＨ３）２ＣＨＯＨ水溶液中Ａｇ ＋ 的光还原及明胶对银颗粒的稳定机理  相似文献   

11.
Definable surface chemistry is essential for many applications of microfluidic polymer systems. However, small cross-section channels with a high surface to volume ratio enhance passive adsorption of molecules that depletes active molecules in solution and contaminates the channel surface. Here, we present a one-step photochemical process to coat the inner surfaces of closed microfluidic channels with a nanometer thick layer of poly(ethylene glycol) (PEG), well known to strongly reduce non-specific adsorption, using only commercially available reagents in an aqueous environment. The coating consists of PEG diacrylate (PEGDA) covalently grafted to polymer surfaces via UV light activation of the water soluble photoinitiator benzoyl benzylamine, a benzophenone derivative. The PEGDA coating was shown to efficiently limit the adsorption of antibodies and other proteins to <5% of the adsorbed amount on uncoated polymer surfaces. The coating could also efficiently suppress the adhesion of mammalian cells as demonstrated using the HT-29 cancer cell line. In a subsequent equivalent process step, protein in aqueous solution could be anchored onto the PEGDA coating in spatially defined patterns with a resolution of <15 μm using an inverted microscope as a projection lithography system. Surface patterns of the cell binding protein fibronectin were photochemically defined inside a closed microfluidic device that was initially homogeneously coated by PEGDA. The resulting fibronectin patterns were shown to greatly improve cell adhesion compared to unexposed areas. This method opens for easy surface modification of closed microfluidic systems through combining a low protein binding PEG-based coating with spatially defined protein patterns of interest.  相似文献   

12.
BackgroundA biosurfactant produced by Pseudomonas aeruginosa cultivated in a low-cost medium formulated with 2.5% vegetable oil refinery residue and 2.5% corn steep liquor and distilled water was employed to stabilize silver nanoparticles in the liquid phase. The particles were initially synthesized using NaBH4 as reducing agent in biosurfactant reverse micelles and were extracted from the micellar solution to disperse in heptane.ResultsA silver particle size in the range of 1.13 nm was observed. The UV–vis absorption spectra proposed that silver nanoparticles could be formed in the reverse micelles and relatively stabilized for at least 3 months without passivator addition. The Transmission Electron Microscope (TEM) shows that the silver nanoparticles are of spherical form and relatively uniform.ConclusionsThis process provided a simpler route for nanoparticle synthesis compared to existing systems using whole organisms or partially purified biological extracts, showing that the low-cost biosurfactant can be used for nanoparticle synthesis as a non-toxic and biodegradable stabilizing agent.  相似文献   

13.
BackgroundRhodotorula glutinis is capable of synthesizing numerous valuable metabolites with extensive potential industrial usage. This paper reports the effect of initial culture medium pH on growth and protein, lipid, and carotenoid biosynthesis by R. glutinis.ResultsThe highest biomass yield was obtained in media with pH 4.0–7.0, and the value after 72 h was 17.2–19.4 gd.w./L. An initial pH of the medium in the range of 4.0–7.0 has no significant effect on the protein (38.5–41.3 g/100 gd.w.), lipid (10.2–12.7 g/100 gd.w.), or carotenoid (191.7–202.9 μg/gd.w.) content in the biomass or on the profile of synthesized fatty acids and carotenoids. The whole pool of fatty acids was dominated by oleic (48.1–53.4%), linoleic (21.4–25.1%), and palmitic acids (13.0–15.8%). In these conditions, the yeast mainly synthesized torulene (43.5–47.7%) and β-carotene (34.7–38.6%), whereas the contribution of torularhodin was only 12.1–16.8%. Cultivation in medium with initial pH 3.0 resulted in a reduction in growth (13.0 gd.w./L) and total carotenoid (115.8 μg/gd.w.), linoleic acid (11.5%), and torularhodin (4.5%) biosynthesis.ConclusionThe different values of initial pH of the culture medium with glycerol and deproteinized potato wastewater had a significant effect on the growth and protein, lipid, and carotenoid biosynthesis by R. glutinis.  相似文献   

14.
A porous silicon (PSi) based microarray has been integrated with a microfluidic system, as a proof of concept device for the optical monitoring of selective label-free DNA-DNA interaction. A 4 × 4 square matrix of PSi one dimensional photonic crystals, each one of 200 μm diameter and spaced by 600 μm, has been sealed by a polydimethylsiloxane (PDMS) channels circuit. The PSi optical microarray elements have been functionalized by DNA single strands after sealing: the microfluidic circuit allows to reduce significantly the biologicals and chemicals consumption, and also the incubation time with respect to a not integrated device. Theoretical calculations, based on finite element method, taking into account molecular interactions, are in good agreement with the experimental results, and the developed numerical model can be used for device optimization. The functionalization process and the interaction between DNA probe and target has been monitored by spectroscopic reflectometry for each PSi element in the microchannels.  相似文献   

15.
The separation of target nucleic acid sequences from biological samples has emerged as a significant process in today''s diagnostics and detection strategies. In addition to the possible clinical applications, the fundamental understanding of target and sequence specific hybridization on surface modified magnetic beads is of high value. In this paper, we describe a novel microfluidic platform that utilizes a mobile magnetic field in static microfluidic channels, where single stranded DNA (ssDNA) molecules are isolated via nucleic acid hybridization. We first established efficient isolation of biotinylated capture probe (BP) using streptavidin-coated magnetic beads. Subsequently, we investigated the hybridization of target ssDNA with BP bound to beads and explained these hybridization kinetics using a dual-species kinetic model. The number of hybridized target ssDNA molecules was determined to be about 6.5 times less than that of BP on the bead surface, due to steric hindrance effects. The hybridization of target ssDNA with non-complementary BP bound to bead was also examined, and non-specific hybridization was found to be insignificant. Finally, we demonstrated highly efficient capture and isolation of target ssDNA in the presence of non-target ssDNA, where as low as 1% target ssDNA can be detected from mixture. The microfluidic method described in this paper is significantly relevant and is broadly applicable, especially towards point-of-care biological diagnostic platforms that require binding and separation of known target biomolecules, such as RNA, ssDNA, or protein.  相似文献   

16.
An event-triggered leader-following consensus problem for multi-agent systems with nonlinear dynamics was investigated in this study. The interaction topologies among the agents that we considered are randomly switched ones, governed by a semi-Markov process with partially unknown rates. By building the state error model between the leader and followers, the consensus problem is first converted into a stability problem. Moreover, an event-triggered transmission scheme based on sampling data was proposed to reduce communication redundancy. The consensus controller and event-triggered parameters can be designed effectively. By constructing a Lyapunov–Krasovskii functional (LKF) with a triple integral, the sufficient conditions required to guarantee the event-triggered consensus can be reached with respect to the linear matrix inequalities (LMIs). Ultimately, the validity of the theoretical results is demonstrated by a numerical example.  相似文献   

17.
BackgroundSynthesis of selenium nanoparticles from selenite by Shewanella sp. HN-41 demonstrated that particle size depended on the reaction time and biomass of cells. The slow reaction and low biomass tended to form small particles. In this study, Shewanella sp. HN-41 was introduced into the anode of a nonexternal circuit bioelectrochemical system (nec_BES) to convert chemical energy from lactate to low electron current to the cathode, where selenite was reduced.ResultsOur experiment with two systems, one bioelectrochemical system with a cathode flushed with nitrogen and the other with a no-nitrogen-flushing cathode, showed that the former could not produce Se nanoparticles after 21 d, but the latter formed them with an average size of 37.7 nm. The SEM and TEM images demonstrated that the particle size of 10 nm occupied over 10% and most of the particles were in the range of 30–60 nm. The XRD result and SAED image demonstrated no clear peaks of crystal and proved that the Se nanoparticles are amorphous.ConclusionsThe clean Se nanoparticles were synthesized and completely separated from bacterial cells in the bioelectrochemical system. This study opened a new approach for the biological synthesis of metal nanoparticles. Finally, the Se products in the range of 30–60 nm can be tested for antimicrobial activities in medical applications.How to cite: Ho CT, Nguyen T-H, Lam T-T, et al. Biogenic synthesis of selenium nanoparticles by Shewanella sp. HN-41 using a modified bioelectrochemical system. Electron J Biotechnol 2021;54. https://doi.org/10.1016/j.ejbt.2021.07.004  相似文献   

18.
We present an immunoaggregation assay chip for multiplexed biomarkers detection. This chip is based on immunoaggregation of antibody functionalized microparticles (Ab-MPs) to quantify concentrations of multiple biomarkers simultaneously. A mixture of multiple types of Ab-MPs probes with different sizes and magnetic properties, which were functionalized by different antibodies, was used for the multiplexed assay. The interactions between biomarkers and their specific Ab-MPs probes caused the immunoaggregation of Ab-MPs. A two-stage micro resistive pulse sensor was used to differentiate and count the Ab-MP aggregates triggered by different biomarkers via size and magnetic property for multiplexed detection. The volume fraction of each type of Ab-MP aggregates indicates the concentration of the corresponding target biomarker. In our study, we demonstrated multiplexed detection of two model biomarkers (human ferritin and mouse anti-rabbit IgG) in 10% fetal bovine serum, using anti-ferritin Ab and anti-mouse IgG Ab functionalized MPs. We found that the volume fraction of Ab-MP aggregates increased with the increased biomarker concentrations. The detection ranges from 5.2 ng/ml to 208 ng/ml and 3.1 ng/ml to 5.12 × 104 ng/ml were achieved for human ferritin and mouse anti-rabbit IgG. This bioassay chip is able to quantitatively detect multiple biomarkers in a single test without fluorescence or enzymatic labeling process and hence is promising to serve as a useful tool for rapid detection of multiple biomarkers in biomedical research and clinical applications.  相似文献   

19.
文章分别考察了日本岛津制作所年轻工程师田中耕一和德国法兰克福大学两位教授上个世纪80年代展开MALDI研究的经纬及其相关研究成果的发表情况。文章指出：尽管双方各自独立地开发出了MALDI法,但前者当年只是以专利的形式,而后者则是以论文的形式公开自己的研究结果,不过,无论是从实验研究,还是从论文发表来看,田中都称得上是使用MALDI法对蛋白质分子展开质谱分析的第一人。文章最后还对田中这位非西方出身,只有大学本科学历,且无SCI论文的企业工程师最终脱颖而出,荣获诺贝尔化学奖一事的意蕴进行了分析。  相似文献   

20.
In an effort to enhance the odds of “organized spontaneity,” this article advances analytics for selection of participants, format of

interaction, and milieu within which the interaction occurs. The analysis proceeds in a two-part process. First, three creative environments far removed from the present-day academic forums—18th-century French salons, the 18th-century London coffeehouses, and Mensa—are examined to generate analytical distance from our current practices. Second, with these three locales in the background, current practices are analyzed and the following thoughts are offered for deliberation: (1) Low threshold to entry is not necessarily a bad thing. (2) Creative environments require an artful mix of homogeneity and heterogeneity. (3) Sociality, which is essential for a creative environment, limits the extent to which a disagreement can be pushed. (4) The selection–format–milieu interaction greatly shapes the character of what gets produced in a creative environment.  相似文献   

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