Rapid simultaneous ultrasensitive immunodetection of five bacterial toxins

Rapid ultrasensitive detection of gastrointestinal pathogens presents a great interest for medical diagnostics and epidemiologic services. Though conventional immunochemical and polymerase chain reaction (PCR)-based methods are sensitive enough for many applications, they usually require several hours for assay, whereas as sensitive but more rapid methods are needed in many practical cases. Here, we report a new microarray-based analytical technique for simultaneous detection of five bacterial toxins: the cholera toxin, the E. coli heat-labile toxin, and three S. aureus toxins (the enterotoxins A and B and the toxic shock syndrome toxin). The assay involves three major steps: electrophoretic collection of toxins on an antibody microarray, labeling of captured antigens with secondary biotinylated antibodies, and detection of biotin labels by scanning the microarray surface with streptavidin-coated magnetic beads in a shear-flow. All the stages are performed in a single flow cell allowing application of electric and magnetic fields as well as optical detection of microarray-bound beads. Replacement of diffusion with a forced transport at all the recognition steps allows one to dramatically decrease both the limit of detection (LOD) and the assay time. We demonstrate here that application of this "active" assay technique to the detection of bacterial toxins in water samples from natural sources and in food samples (milk and meat extracts) allowed one to perform the assay in less than 10 min and to decrease the LOD to 0.1-1 pg/mL for water and to 1 pg/mL for food samples.     

超声空化对喇曼散射的影响

1　引言超声空化的过程是集中声场能量并迅即释放的过程。空化泡崩溃时 ,极短时间在空化泡周围的极小空间内 ,产生 50 0 0 K以上的高温和大约 5× 10 7Pa的高压 ,温度变化率高达 10 9K/ s,并伴生强烈的冲击波和时速达 4 0 0 km的射流 [1]。在这样特殊的物理环境下 ,物质的物理性质和一些物理现象是否发生变化或受环境的影响呢 ?作者曾经设计了一种特殊形状的喇曼光谱样品池 ,它既能让激光束透过样品 ,又能在样品中引入超声空化场 ,从而可研究超声空化对喇曼散射光谱的影响 [2 ]。作为该研究工作的继续 ,本文选择了四氯化碳和乙二醇两种样…     

超声空化效应对溶液电导率的影响

大功率超声作用于溶液时会产生空化效应,并影响溶液的物理化学性质,如电导率、液体粘度及液体表面张力等。文章对不同功率的超声作用于弱电解质溶液时其电导率的变化进行了实验研究,发现溶液电导率与超声波强度有关。当强度增大到有空化产生时,电导率开始减小;继续增大强度,溶液电导率减小到一定程度后有小幅回升现象出现。且液体内空化效应的出现将导致液体的电导率减小,空化达到一定强度时,空化效应引起的局部瞬态高温高压、冲击波和微射流导致的新导电粒子,会使电导率小幅回升。     

超声聚能器空化腐蚀的研究进展

空化腐蚀是影响超声聚能器工作效率和使用寿命的重要因素,研究超声聚能器的空化腐蚀具有重要意义。分析了超声聚能器材质、辐射面粗糙度、材料的微观组织结构和力学性能对其抗空化腐蚀的影响,探讨了抑制空化腐蚀的相关措施。     

相界面上超声空化气泡聚并、滑移促进的传质

根据超声空化泡在相间特殊运动时周围流体流动特性,以相间传质渗透理论为依据,结合流体动力学原理,在充分研究相界面上气泡行为的基础上,根据界面上超声空化气泡生长过程的动力学行为,导出了相界面上超声空化气泡聚并和滑移促进的传质模型。在此基础上,建立了相界面上气泡滑移、聚并及脱离界面逸出这三种气泡行为共同促进的传质模型,该模型可用于描述超声空化界面等相界面上的传质行为,为超声波强化传质过程提供了理论依据。     

非同频水平正交超声束的声化学效应

本文采用碘释放法观察到两个非同频超声束水平正交幅照的声化学产额能获得明显的增强效应     

Controlled effect of ultrasonic cavitation on hydrophobic/hydrophilic surfaces

Controlling cavitation at the solid surface is of increasing interest, as it plays a major role in many physical and chemical processes related to the modification of solid surfaces and formation of multicomponent nanoparticles. Here, we show a selective control of ultrasonic cavitation on metal surfaces with different hydrophobicity. By applying a microcontact printing technique we successfully formed hydrophobic/hydrophilic alternating well-defined microstructures on aluminium surfaces. Fabrication of patterned surfaces provides the unique opportunity to verify a model of heterogeneous nucleation of cavitation bubbles near the solid/water interface by varying the wettability of the surface, temperature and ultrasonic power. At the initial stage of sonication (up to 30 min), microjets and shock waves resulting from the collapsing bubbles preferably impact the hydrophobic surface, whereas the hydrophilic areas of the patterned Al remain unchanged. Longer sonication periods affect both surfaces. These findings confirm the expectation that higher contact angle causes a lower energy barrier, thus cavitation dominates at the hydrophobic surfaces. Experimental results are in good agreement with expectations from nucleation theory. This paper illustrates a new approach to ultrasound induced modification of solid surfaces resulting in the formation of foam-structured metal surfaces.     

Tensile strength and visible ultrasonic cavitation of superfluid4He

Motion picture photographs of cavitation in He II revealed new characteristics pertinent to the liquid's tensile strength and bubble dynamics. A cylindrical acoustic standing wave with a frequency of 50.58 kHz induced the cavitation in He II at a temperature of 2.09 K. Analysis of light diffracted by the sound gave measurements of the acoustic pressure amplitude which were used both for selecting the best drive frequency and for obtaining the tensile strength. Bubbles appeared to originate on pressure antinodes, expanded to a diameter of 0.5–1.0 mm in about 0.3 msec, and eventually fragmented into smaller bubbles. They originated where the negative pressure extremum was as small as –0.6 bar (+0, –50%), a tensile strength much smaller than the predictions of theories developed for the homogeneous nucleation of bubbles in classical liquids. The bubble fragments were frequently nonspherical and had widths of 0.1–0.2 mm. Small bubbles also displayed an unexpected preference to originate on the surface of a stainless steel tube inserted in the sound field. Subsequent to nucleation, bubbles were frequently attracted to acoustic pressure nodes in agreement with a theory of vibrations and forces originally developed for bubbles in normal liquids. Attempts to detect first and second sound radiated by cavitation are described.Work supported by the National Science Foundation Grant DMR75-15628.NSF Predoctoral Fellow.     

Features of cavitation and cavitation erosion of waveguides of powerful ultrasonic units at higher-than-average pressure of the media

The features of the acoustic effects of cavitation created by the rod waveguides of powerful ultrasonic magnetostriction units in different liquid media at a pressure to 5 MPa have been investigated. It has been established that, under the conditions of maximum acoustical activity of cavitation realized at the medium's higher-than-average pressure, the erosion damage to the waveguides is of an unusual structurized character and develops from the central part of the end to its periphery. A result of the erosion damage may be the total loss of operating capacity by the waveguides manufactured from the most erosion-resistant materials (stainless steel and titanium alloys) even after a few hours of tests. Translated from Inzhenerno-Fizicheskii Zhurnal, Vol. 81, No. 4, pp. 690–696, July–August, 2008.     

Application of an ultrasonic technique to creep cavitation in silicon nitride

A non-destructive method based on measurements of ultrasonic wave velocities and Young's modulus is proposed for quantification of creep cavitation in silicon nitride. Tensile creep tests of silicon nitride were conducted at 1400°C in air and the tests were periodically interrupted to measure the longitudinal and transverse ultrasonic wave velocities and Young's modulus. The velocities and Young's modulus decreased linearly with tensile creep strain. The volume fraction of cavities was estimated from the values of the ultrasonic wave velocities and Young's modulus, and compared with the cavity volume predicted from tensile creep strain. The dependence of Young's modulus on volume fraction of cavities is discussed.     

The evolution of a cavitation zone in a focused ultrasonic field

We have experimentally studied the generation of sonoluminescence (SL) and cavitation noise in the field of a focusing ultrasonic radiator during a gradual smooth increase in the applied voltage. In addition to the SL signal, we have recorded the output signal of a hydrophone situated behind the focal region of the radiator and measured the cavitation-noise spectrum. Four stages in the development of a cavitation zone have been distinguished as manifested by the specific character of dependences of the measured parameters on the voltage applied to the radiator. Spectral signs of the cavitation-noise characteristic of each stage of development of the cavitation zone.     

超声空化对溴化锂溶液气泡运动的影响

根据溴化锂溶液的物性参数方程,利用试验数据和超声作用于均相液体中空化气泡运动的动力学模型,推导出溴化锂溶液超声空化气泡运动方程。利用MATLAB对溴化锂溶液超声空化气泡的运动特性进行数值模拟,对过冷沸腾和饱和沸腾条件下溴化锂溶液沸腾气泡的运动变化特性进行研究,得到超声空化效应对气泡运动特性变化影响的模拟结果。为溴化锂溶液传热性能和吸收式制冷机组制冷效率的提高提供一定的理论依据。     

Destruction of bacterial spores by phenomenally high efficiency non-contact ultrasonic transducers

Conventional wisdom stipulates that high power ultrasound without direct or indirect transducer contact with the medium to be treated is not possible. This seemingly correct notion is based upon two major hurdles: inefficient transmission of ultrasound from the piezoelectric material into air/gases and exorbitant attenuation of ultrasound by gases. The latter is a natural phenomenon about which nothing can be done, and the former requires an un-conventional approach to transducer design. After many years of R& D in this area, we have finally succeeded in producing transducers that generate immense acoustic pressure in air in the frequency range of ∼50 kHz→10 MHz. By using these transducers without any contact with the material, we demonstrate destruction of 99.9% of dried bacterial spore samples of a close relative of anthrax, Bacillus thuringiensis. Following further refinement of the transducers and the mechanism of their excitation, we anticipate that non-contact ultrasound will have numerous applications including inactivation of agents of bioterrorism and sterilization of medical and surgical equipment, food materials, and air-duct systems of buildings, airplanes, space stations, and others. Electronic Publication     

基于共振峰的超声空化声发射信号分析

摘 要：为了简单有效地提取超声空化时频信息而估计空化强度,提出了应用基于全极点线性预测编码模型的共振峰分析方法进行分析,并使用其分析了19kHz超声作用下的空化场的声发射信号。将共振峰分析结果与常用的频谱分析和短时傅利叶分析结果进行了对比,发现共振峰分析方法更能从超声空化声发射信号的中提取出简单但关键的时频信息来。采用共振峰方法的分析结果表明,可以利用空化声发射的共振峰参数如基频频率和相对高频共振峰的幅值来分别反映超声空化的非线性振荡和瞬态空化,进而估计超声空化的强度。     

柱形料腔中超声空化效应的空间分布特性研究

研究了20 kHz的圆柱形料腔中超声空化效应的形成及其空间分布特性。应用柱贝塞尔函数,推导获取了柱形声场内超声传播的声能密度的分布,并采用有限元方法进行仿真分析。针对频率为20 kHz的功率超声实验,结合声学测量方法和鲁米诺声致化学发光方法,对理论分析结果进行了验证对照。结果表明:料腔半径R=50 mm,20 kHz谐振液位高度H=90 mm时,若功放电流<40 mA,超声空化效应出现在变幅杆端部区域;若40 mA≤功放电流≤80 mA,空化效应显著增强,空化效应的空间分布与场内声压分布一致,空化效应受声模态影响,形成远场空化效应的分布特性;若功放电流>80 mA,受非线性因素影响,谐振液位时,空化效应在声流作用下呈柱形拖尾状分布,并在底部壁面边界形成平铺状分布;非谐振液位高度等于75 mm时,超声空化效应随功率增加仅在变幅杆端部区域出现,且呈现局域空化分布特性。     

Bacillus globigii bugbeads: a model simulant of a bacterial spore

Nonpathogenic microorganisms are often used as simulants of biological pathogens during the initial phase of detection method development. While these simulants approximate the size, shape, and cellular organization of the microorganism of interest, they do not resemble its surface protein content, a factor particularly important in methods based on immunorecognition. Here, we develop and detect an artificial bacterial spore--B. globigii (BG) Bugbead-a particle mimicking the antigenic surface of BG spores. Two methods of spore protein extraction were compared both quantitatively (by protein concentration assay) and qualitatively (by SDS-PAGE and Western blot): extraction by mechanical disruption and extraction by chemical decoating. The former method was more efficient in producing more protein and a greater number of antigens. BG Bugbeads were made by conjugating the extracted proteins to 0.8-microm carboxyl-coated polystyrene particles via carbodiimide coupling. BG Bugbeads were successfully detected by a bead-based enzyme-labeled immunoassay with fluorescence detection with a detection limit of 6.9 x 10(3) particles/mL. Formation of the Bugbead-capture bead complex was confirmed by ESEM. The concept of a harmless artificial spore can be applied to developing improved simulants for pathogenic spore-forming microorganisms such as B. anthracis, C. botulinum, and B. cereus, which can to be used for method validation, instrument calibration, and troubleshooting.     

声流条件下超声空化气泡分布研究

研究了频率为20 kHz的超声作用在圆柱形料腔中出现声流现象时超声空化效应的空间分布特性。结合大振幅声源条件下的声辐射力,对声场内的声流现象进行了仿真分析,获取了不同超声功率和液位高度下的声流速度场分布,初步探究了声流条件下空化气泡的运动分布规律。采用超声空化效应的声致化学发光实验,对比研究了有、无声流条件时超声空化效应的空间分布特性。结果表明：功放电流高于80 mA(电功率为17.6 W)时,超声场可形成稳定的声致流动现象且可有效提高其声能辐射效率,大大增加了空化效应的作用区域,进而提高了声化学反应效率;声流条件下料腔内超声空化效应的分布区域与超声功率(振幅)、料腔液位高度相关,功放电流从40 mA(电功率为8.8 W)增加至120 mA(电功率为26.4 W)时,空化面积占比提高了100.86%,液位高度为60 mm时的空化面积占比较50 mm和70 mm时分别提高了13.11%和73.91%,提高超声功率及选择合理的料腔液位高度,可有效提高空化气泡扩散距离,增大空化分布面积;对于固定形状及尺寸料腔中的声场,声流速度达到一定阈值时,会出现空化效应增强,空化效应增强区域位于大于声流速...     

基于非线性超声空化效应的铝合金热浸镀工艺

在390℃热浸镀锌过程中施加20 kHz的超声波,采用数值分析法求解Keller-Miksis和Mettin方程,描述ZnAl8熔池中0～800 W超声空泡的生长规律和空化效应,研究超声功率对镀层ZnAl8合金组织的影响,以及对1050铝合金表面氧化膜的作用.结果表明:空化效应与超声功率呈现出非线性的变化规律,当超声功率为0～500 W时,空化以稳态效应为主,空化能量不足以消除镀层合金初生相的粗大枝晶组织,也不能消除铝合金氧化膜.当功率为600～800 W时,空化以瞬态崩溃破裂的方式释放能量,其中700～800 W的空化压强和温度效应可以将铝合金表面氧化膜击碎和熔蚀,为镀层中元素扩散提供了铺展润湿和物质传输的通道,镀层ZnAl8合金与1050铝合金基体之间形成了良好的冶金结合,镀层合金呈细小均匀的蔷薇组织.     

Effect of electroceramic particles on damping behaviour of aluminium hybrid composites produced by ultrasonic cavitation and mechanical stirring

In this study, electroceramics PBN and PLZT along with SiC were included in Al–3.96 wt.% Mg (A514.0) master alloy. Ultrasonic cavitation (UST) and mechanical stirring (MS) were employed to improve wettability and dispersion during casting. Two composite systems were produced: PBN system (5 wt.% PBN + 1 wt.% SiC and 15 wt.% PBN + 1 wt.% SiC) and the PLZT system (follows the same designation). The influence of fabrication method on the microstructures, particle distribution and wettability as well as electroceramic impact on dynamo-mechanical properties of prepared composites were investigated. Optical microscope (OM) and scanning electron microscope (SEM) results indicate that the processing technique was effective as it promoted wettability and homogeneous dispersion of particles throughout the Al matrix. Dynamic mechanical analysis (DMA) study of the composites demonstrated that the addition of the functional particles to the Al alloy matrix improved damping capacity (Tan δ) at 200 °C. The composites exhibited an increase in Tan δ of 24.3 ± 0.3% and 91.4 ± 0.2% for 5 and 15 wt.% PBN + 1 wt.% SiC and an increase of 19.7 ± 0.5% and 42.5 ± 0.3% for 5 and 15 wt.% PLZT + 1 wt.% SiC, respectively, when compared to the aluminium alloy matrix.     

In situ surface-etched bacterial spore detection using dipicolinic acid-europium-silica nanoparticle bioreporters

A basic approach was optimized for the synthesis of highly selective and sensitive in situ mesoporous (MCM) type imprinted silica polymers for the detection of dipicolinic acid (DPA) using europium as a reporter. DPA is a ubiquitous biochemical marker available during the germination event of endospore-forming bacteria such as Bacillus . Additionally, an MCM-MIP (molecularly imprinted polymeric phenomena) detector and a companion MCM-non-surface-MIP detector were synthesized using europium reporters for the sensing of DPA under optimized laboratory conditions. Our results showed that the in situ molecular imprinting process enabled rapid, selective detection of DPA with high sensitivity compared to MCM-MIP (imprinted for DPA; no DPA present), MCM-Non-MIP (no imprint present), and MCM-SR-MIP (imprinted with DPA present) detectors. The lower detection limit observed for DPA concentration is 5.49 × 10(-10) mol dm(-3) for MCM-MIP. The performance of the sensor in high-salt-water conditions, under photo-bleaching, and its reusability were also evaluated. The synthesized in situ MCM-MIP material should permit the detection of DPA for field assays related to suspect bacterial sporulation events.