Protein separation and identification using magnetic beads encoded with surface-enhanced Raman spectroscopy

This article presents a prototype of a surface-enhanced Raman spectroscopy (SERS)-encoded magnetic bead of 8 μm diameter. The core part of the bead is composed of a magnetic nanoparticle (NP)-embedded sulfonated polystyrene bead. The outer part of the bead is embedded with Ag NPs on which labeling molecules generating specific SERS bands are adsorbed. A silica shell is fabricated for further bioconjugation and protection of SERS signaling. Benzenethiol, 4-mercaptotoluene, 2-naphthalenethiol, and 4-aminothiophenol are used as labeling molecules. The magnetic SERS beads are used as substrates for protein sensing and screening with easy handling. As a model application, streptavidin-bound magnetic SERS beads are used to illustrate selective separation in a flow cytometry system, and the screened beads are spectrally recognized by Raman spectroscopy. The proposed magnetic SERS beads are likely to be used as a versatile solid support for protein sensing and screening in multiple assay technology.     

Electrochemical DNA biosensor based on silver nanoparticles/poly(3-(3-pyridyl) acrylic acid)/carbon nanotubes modified electrode

In this work, we present an electrochemical DNA sensor based on silver nanoparticles/poly(trans-3-(3-pyridyl) acrylic acid) (PPAA)/multiwalled carbon nanotubes with carboxyl groups (MWCNTs-COOH) modified glassy carbon electrode (GCE). The polymer film was electropolymerized onto MWCNTs-COOH modified electrode by cyclic voltammetry (CV), and then silver nanoparticles were electrodeposited on the surface of PPAA/MWCNTs-COOH composite film. Thiol group end single-stranded DNA (HS-ssDNA) probe was easily covalently linked onto the surface of silver nanoparticles through a 5′ thiol linker. The DNA hybridization events were monitored based on the signal of the intercalated adriamycin by differential pulse voltammetry (DPV). Based on the response of adriamycin, only the complementary oligonucleotides gave an obvious current signal compared with the three-base mismatched and noncomplementary oligonucleotides. Under the optimal conditions, the increase of reduction peak current of adriamycin was linear with the logarithm of the concentration of the complementary oligonucleotides from 9.0 × 10⁻¹² to 9.0 × 10⁻⁹ M with a detection limit of 3.2 × 10⁻¹² M. In addition, this DNA sensor exhibited an excellent reproducibility and stability during DNA hybridization assay.     

血雉的产卵时间和产卵间隔

鸟类的产卵时间和产卵间隔因种而异 ,并会影响其繁殖成功。采用在巢内放置含有温度数据自动记录装置的假卵 ,通过记录其温度的变化来监测雌鸟产卵行为的方法 ,对分布于我国青藏高原森林鸟类血雉(Ithaginiscruentus)的产卵时间和产卵间隔进行了研究。结果表明 ,雌鸟每次入巢产卵的时间是在 15 :16± 2 2min (13 :30～ 18:5 1) ,产卵高峰集中在 14:0 0～ 16 :0 0 ,每次在巢中停留 (2 2± 0 2 6 )h。产卵间隔为 (46 4± 3 0 8)h ,大多数 (75 % )产卵间隔 45 8～ 48 9h。由此推测 ,血雉产卵时间较为集中很可能与避开夜行性捕食者的活动高峰有关 ;而隔天产卵则是对高原恶劣环境条件的适应。     

长江水系钱草鱼遗传结构及变异性的RAPD研究

采用随机扩增多态性DNA（RAPD）技术对长江中游的湖北嘉鱼与江西瑞昌两个地理群体和中游的汉江和湘江两大水系的鲢和草鱼的群体进行了遗传学研究。发现长江水系链的遗传变异要高于草鱼,与现今生物量成反比的反常现象。鲢遗传多样性从大到小的分布是嘉鱼→湘江→瑞昌→汉江。草鱼遗传多样性从大到小的分布是瑞昌→汉江→湘江→嘉鱼,鲢和草鱼的遗传多样性地理分布并不一致。遗传分化指数Gst分别为12.3%和17.5%,表明鲢和草鱼的四个地理群体的遗传分化度较低,可能与地理较近和基因交流频繁有关。     

Core–shell silver nanoparticles for optical labeling of cells

Silver nanoparticles have been modified with self-assembled monolayers of hydroxyl-terminated long chain thiols and encapsulated with a silica shell. The resulting core–shell nanoparticles were used as optical labels for cell analysis using flow cytometry and microscopy. The excitation of plasmon resonances in nanoparticles results in strong depolarized scattering of visible light, permitting detection at the single nanoparticle level. The nanoparticles were modified with neutravidin via epoxide–azide coupling chemistry, to which biotinylated antibodies targeting cell surface receptors were bound. The nanoparticle labels exhibited long-term stability in solutions with high salt concentrations without aggregation or silver etching. Labeled cells exhibited two orders of magnitude enhancement of the scattering intensity compared with unlabeled cells.     

Anti-cancer activity of biosynthesized silver nanoparticles using Avicennia marina against A549 lung cancer cells through ROS/mitochondrial damages

The biosynthesized Ag NPs was synthesized by using marine mangrove plant extract Avicennia marina. The synthesized Ag NPs was confirmed by various physiochemical characterization including UV-spectrometer and XRD analysis. In addition, the shape and of the synthesized Ag NPs was morphologically identified by SEM initially and TEM finally. After confirmation, the anti-cancer property of synthesized Ag NPs was confirmed at 50 µg/mL concentration against A549 lung cancer cells by MTT assay. Further, the ability to stimulate the ROS generation and mitochondrial membrane at the IC₅₀ concentration of Ag NPs was confirmed by fluorescence microscopy using DCFH-DA and rhodamine 123 dyes respectively. Finally, the result was concluded that the synthesized Ag NPs has improved anti-cancer activity against A549 cells at lowest concentration.     

Effect of treatment with silver(I) complex of metronidazole on ocular rosacea: Design and formulation of new silver drug with potent antimicrobial activity

BackgroundMetronidazole with well-soluble silver nitrate forms a complex with potent activity (synergism) against bacterial strains and fungi with simultaneous lower side effect than in the case of the two agents administered separately.This study aimed to establish effectiveness in the treatment of ocular rosacea with new formulation of drops and ointment with silver(I) complex of metronidazole.MethodsThree patients suffering from serious ophthalmic complications of acne rosacea were treated with drops and ointments applied 3 times a day for 3 months. The uncorrected visual acuity testing (UCVA), pachymetry as well as subjective and objective tear film assessment were evaluated.ResultsIn all patients, we have improved UCVA as well as objective and subjective evaluation of the tear film parameters. No significant differences in corneal thickness were observed.ConclusionWell soluble silver(I) complex of metronidazole might be an alternative method in ophthalmic complications of acne rosacea treatment.     

An intracellular silver deposition method for targeted detection and chemical analysis of uncultured microorganisms

The vast majority of environmental bacteria remain uncultured, despite two centuries of effort in cultivating microorganisms. Our knowledge of their physiology and metabolic activity depends to a large extent on methods capable of analyzing single cells. Bacterial identification is a key step required by all currently used single-cell imaging techniques and is typically performed by means of fluorescent labeling. However, fluorescent cells cannot be visualized by ion- and electron microscopy and thus only correlative, indirect, cell identification is possible. Here we present a new method of bacterial identification by in situ hybridization coupled to the deposition of elemental silver nanoparticles (silver-DISH). We show that hybridized cells containing silver can be directly visualized by light microscopy, scanning electron microscopy, energy dispersive X-ray spectroscopy, secondary ion mass spectrometry (nanoSIMS), and confocal Raman micro-spectroscopy. Silver-DISH did not alter the isotopic (¹³C) and elemental composition of stable-isotope probed cells more than other available hybridization methods, making silver-DISH suitable for broad applications in stable-isotope labeling studies. Additionally, we demonstrate that silver-DISH can induce a surface-enhanced Raman scattering (SERS) effect, amplifying the Raman signal of biomolecules inside bacterial cells. This makes silver-DISH the only currently available method that is capable of delivering a SERS-active substrate inside specifically targeted microbial cells.     

Rice leaf extract synthesized silver nanoparticles: An in vitro fungicidal evaluation against Rhizoctonia solani,the causative agent of sheath blight disease in rice

Silver nanoparticles (Ag NP) were synthesized using rice leaf extract and optimized synthetic conditions were found to be 0.4 % leaf extract, 0.6 mM AgNO₃ and 30 min of autoclaving. Produced NP were characterized using UV–vis, DLS, zeta potential, XRD, TEM and FTIR. Ag NP formation was established from UV–vis spectra and NP showed zeta potential value of −27.4 mV. NP were spherical, polydisperse and average size was 16.5 ± 6.2 nm. Antifungal activity of Ag NP was assessed by poisoned food technique and resazurin broth dilution against mycelium and sclerotia of fungus R. solani, the causative agent of sheath blight disease in rice. Results confirmed effective hyphal growth inhibition and % growth inhibition was dose dependent (2.5–10 μg/mL). Ag NP showed enhanced mycelial inhibition (81.7–96.7 %) at 10 μg/mL. MIC values of Ag NP were in the range of 5–10 and 15–20 μg/mL towards fungal mycelium and sclerotia, respectively. Ag NP treatment (20 μg/mL) completely inhibited the disease incidence at 20 μg/mL. Ag NP treatment (10 μg/mL) caused 1.3 and 1.5 times enhancement in seedling vigor index. Hence, Ag NP can be utilized towards management and control of various fungal diseases of crops.