基于DNA分子导线的纳米生物传感器

DNA分子导线具有独特的导电性能和塞贝克(Seebeck)效应,它是构筑电化学纳米生物传感器和热电偶生物传感器的理想材料。文章简要介绍了DNA分子导线的制备方法及导电机理,以及基于DNA分子导线的纳米生物传感器的分子识别机制,着重分析了基于DNA分子导线的纳米生物传感器的传感原理。文章还介绍了基于DNA分子导线的纳米生物传感器在基因分析、单碱基突变检测等方面的应用。     

50-Hz electromagnetic environment and the incidence of childhood tumors in Stockholm County

The magnetic fields from overhead power lines and other electromagnetic sources were determined at the birth and diagnosis dwellings of all tumor cases reported in the county of Stockholm during the years 1958-73 for individuals 0-18 years of age. The study was limited to 716 cases having a permanent address in the county both at time of birth and diagnosis. An equivalent number of controls was matched to the cases according to church district of birth, age, and sex. Outside each dwelling, the occurrence of visible electrical constructions (6-200-kV high-voltage wires, substations, transformers, electric railroads, and subways) within 150 m of the dwelling was noted. Also, the 50-Hz magnetic field was measured outside the main entrance of the dwelling. Visible 200-kv wires were noted at 45 of 2,098 dwellings and were found twice as frequently among cases as among controls (P less than .05). The magnetic field measured at the dwelling varied between 0.0004 to 1.9 microT (mean value 0.069 microT). The magnetic field was higher (0.22 microT) at dwellings with visible 200-kV wires than at those without such wires. Magnetic fields of 0.3 microT or more were measured at 48 dwellings, and were found twice as frequently among cases as among controls (P less than .05). The difference was most pronounced for dwellings of nervous system tumors and was less for leukemias.     

Crystal structure of phosphatidylglycerophosphatase (PGPase), a putative membrane-bound lipid phosphatase, reveals a novel binuclear metal binding site and two "proton wires"

Phosphatidylglycerophosphatase (PGPase), an enzyme involved in lipid metabolism, catalyzes formation of phosphatidylglycerol from phosphatidylglycerophosphate. Phosphatidylglycerol is a multifunctional phospholipid, found in the biological membranes of many organisms. Here, we report the crystal structure of Listeria monocytogenes PGPase at 1.8 A resolution. PGPase, an all-helical molecule, forms a homotetramer. Each protomer contains an independent active site with two metal ions, Ca(2+) and Mg(2+), forming a hetero-binuclear center located in a hydrophilic cavity near the surface of the molecule. The binuclear center, conserved ligands, metal-bound water molecules, and an Asp-His dyad form the active site. The catalytic mechanism of this enzyme is likely to proceed via binuclear metal activated nucleophilic water. The binuclear metal-binding active-site environment of this structure should provide insights into substrate binding and metal-dependent catalysis. A long channel with inter-linked linear water chains, termed "proton wires," is observed at the tetramer interface. Comparison of similar water chain structures in photosynthetic reaction centers (RCs), Cytochrome f, gramicidin, and bacteriorhodopsin, suggests that PGPase may conduct protons via proton wires.     

应用缝合锚结合联合腱外侧半肌腱重建喙锁韧带治疗肩锁关节脱位

目的:观察治疗肩锁关节脱位的不同手术方式的治疗效果,以确定最佳的手术方法。方法:将60例肩锁关节脱位病人随机分为治疗组和对照组,分别采用缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动的治疗方法及克氏针张力带内固定方法,术后3个月及6个月对患者进行JOA总体情况评定、JOA评分比较、肩锁关节疼痛VAS评定情况比较及安全性评价。结果:应用缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动治疗肩锁关节脱位3个月及6个月后JOA总体评定情况结果及.JOA评分结果优于克氏针张力带治疗方法;应用缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动治疗肩锁关节脱位3个月及6个月后肩锁关节疼痛程度均轻于同时期克氏针张力带治疗方法。结论:缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动治疗肩锁关节脱位是一种安全有效的临床治疗方法。     

DNA frayed wires: Differential polymerization of d(AnGm) oligonucleotides

Oligodeoxyribonucleotides with terminal runs of contiguous guanines, d(A_nG_m), spontaneously associate into high molecular weight complexes that resolve on polyacrylamide gels as a regular ladder pattern of bands with low mobility. The aggregates, which we call frayed wires, arise from the interaction between the guanine residues of the oligonucleotides; the adenine tracts are single stranded and can take part in Watson–Crick interactions. Oligonucleotides, with different arm‐to‐stem ratios and total length, readily associate in the presence of Mg²⁺ to form aggregates consisting of an integer number of strands. The type of the observed aggregates is determined by the length of the guanine run. Oligonucleotides with six guanines form four‐ and eight‐stranded complexes; there is no further polymerization. An increase in the number of guanine residues to 10 and 15 leads to polymerization resulting in a ladder pattern of up to 9 bands and an intense signal at the top of the gel. The relative population of any given species in a frayed wire sample is governed by the guanine stem length and is not affected to any substantial extent by arms up to 40 bases long. The type and concentration of the cation in the solution affect the degree of aggregation, with Na⁺ and K⁺ promoting the formation of complexes comprised of 2–4 strands and Mg²⁺ being the most effective in facilitating polymerization. The electrophoretic behavior of frayed wires was analyzed in the framework of the Ogston theory. The free mobility of frayed wires in the solution is close to the values reported for single‐stranded DNA, indicating the equivalence of the charge density of the two conformations. The retardation coefficients for frayed wires arising from a single kind of parent strand increase with the introduction of each additional strand. There is no correlation between the retardation coefficient and the type of parent strand; rather, the magnitude of the retardation coefficient is determined by the total molecular weight of the complex. The values of the retardation coefficients are consistently higher than those for double‐stranded DNA and they display much stronger dependence on the total molecular weight. Presumably, the distinct structural and dynamic characteristics of the two conformations account for their different electrophoretic behavior. © 1999 John Wiley & Sons, Inc. Biopoly 49: 287–295, 1999     

A study of the interactions that stabilize DNA frayed wires

Oligodeoxyribonucleotides (ODNs) with long, terminal runs of consecutive guanines, and either a dA or dT tract at the other end form higher-order structures called DNA frayed wires. These aggregates self-assemble into species consisting of 2, 3, 4, 5, … associated strands. Some of the remarkable features of these structures are their extreme thermostability and resistance to chemical denaturants and nucleases. However, the nature of the molecular interactions that stabilize these structures remains unclear. Based on dimethyl sulfate (DMS) methylation results, our group previously proposed DNA frayed wires to be a unique set of nucleic-acid assemblies in which the N7 of guanine does not participate in the guanine–guanine interactions. To probe the hydrogen bonding involved in the stabilization of d(A₁₅G₁₅) frayed wires, we used Raman spectroscopy in which the DNA sample is held in photonic crystal fibers. This technique significantly enhances the signals thus allowing the use of very low laser power. Based on our results for d(A₁₅G₁₅) and those of incorporating the isoelectronic guanine analog pyrazolo[3,4,-d]pyrimidine or PPG, into a frayed wire-forming sequence, we provide evidence that these structures are based on the G-quadruplex model. Furthermore, from the Raman spectrum, we observed markers that are consistent with the presence of deoxyguanosine residues in the syn conformation, this suggests the presence of anti-parallel G-quadruplexes. To identify the species that contain syn guanine residues, we used circular dichroism and gel electrophoresis to study an ODN in which all of the guanine residues were brominated, d(A₁₅^8-BrG₁₅). In the presence of potassium, d(A₁₅^8-BrG₁₅) forms what appears to be an anti-parallel dimeric G-quadruplex. To our knowledge, this is the first report of a DNA sequence having all its guanine residues replaced by 8-bromo-guanine and maintaining its ability to form a G-quadruplex structure.     

应用缝合锚结合联合腱外侧半肌腱重建喙锁韧带治疗肩锁关节脱位

目的：观察治疗肩锁关节脱位的不同手术方式的治疗效果,以确定最佳的手术方法。方法：将60例肩锁关节脱位病人随机分为治疗组和对照组,分别采用缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动的治疗方法及克氏针张力带内固定方法,术后3个月及6个月对患者进行JOA总体情况评定、JOA评分比较、肩锁关节疼痛VAS评定情况比较及安全性评价。结果：应用缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动治疗肩锁关节脱位3个月及6个月后JOA总体评定情况结果及JOA评分结果优于克氏针张力带治疗方法;应用缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动治疗肩锁关节脱位3个月及6个月后肩锁关节疼痛程度均轻于同时期克氏针张力带治疗方法。结论：缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动治疗肩锁关节脱位是一种安全有效的临床治疗方法。     

Utilizing Waste Cable Wires for High‐Performance Fiber‐Based Hybrid Supercapacitors: An Effective Approach to Electronic‐Waste Management

In recent years, electronic waste (e‐waste) such as old cable wires, fans, circuit boards, etc., can be often seen in large piles of leftover in dumping yards. Employing these e‐waste sources for energy storage devices not only increases the economic value but also decreases the reliance on fossil fuels. In this context, waste cable wires are utilized to obtain precious copper (Cu) fibers and used as a cost‐effective current collector for the fabrication of fiber‐based hybrid supercapacitor (FHSC). With the braided Cu fibers, forest‐like nickel oxide nanosheet grafted carbon nanotube coupled copper oxide nanowire arrays (NiO NSs@CNTs@CuO NWAs/Cu fibers) are designed via simple wet‐chemical approaches. As a battery‐type material, the forest‐like NiO NSs@CNTs@CuO NWAs/Cu fiber electrode shows superior electrochemical properties including high specific capacity (230.48 mA h g^?1) and cycling stability (82.72%) in aqueous alkaline electrolyte. Moreover, a solid‐state FHSC is also fabricated using forest‐like NiO NSs@CNTs@CuO NWAs/Cu fibers as a positive electrode and activated carbon coated carbon fibers as a negative electrode with a gel electrolyte, which also shows a higher energy and power densities of 26.32 W h kg^?1 and 1218.33 W kg^?1, respectively. The flexible FHSC is further employed as an energy source for various electronic gadgets, demonstrating its suitability for wearable applications.     

In vivo disposition and stability of DNA frayed wires in mice

DNA frayed wires (DNA_FW) are an alternate form of DNA organization formed by the self-association of several strands of guanine-rich oligonucleotides. The purpose of this study was to define for the first time the blood clearance kinetics, tissue distribution, and stability of DNA_FW in vivo in mice. Single bolus doses (1200 pmol/mouse) of ³²P-DNA_FW and ³²P-random DNA were administered intravenously (IV) and intraperitoneally (IP) followed by scheduled blood, urine, fecal and tissue samplings. Blood clearance kinetics was described well by a first order two-compartment open model. The overall half-lives of elimination from the central compartment (T_1/2)_K10 were 3.57 ± 0.1 h for IV and 2.38 ± 0.11 h for IP. In contrast, random DNA was completely degraded after 15 min regardless of the route of administration. Tissue distribution results demonstrated that DNA_FW were primarily distributed and retained in the liver, intestines, kidneys, and heart. Low levels could also be detected in brain. Autoradiographs of blood, tissues, feces and urine extracts established that DNA_FW remained intact after administration as no measurable levels of metabolites or degradation products were found after 24 h. ³²P-DNA_FW was primarily eliminated via hepato-biliary excretion into feces after either IV or IP administration (51.8 ± 4.53% and 36.2 ± 3.4%, respectively). The improved stability and longer half-life of DNA_FW, previously shown in vitro, is also seen in vivo, indicating that DNA_FW may provide a stable delivery system for DNA gene therapies. In conclusion, this is the first study demonstrating the in vivo stability, pharmacokinetics, and disposition of DNA superstructures.     

Analysis of proton wires in the enzyme active site suggests a mechanism of c‐di‐GMP hydrolysis by the EAL domain phosphodiesterases

We report for the first time a hydrolysis mechanism of the cyclic dimeric guanosine monophosphate (c‐di‐GMP) by the EAL domain phosphodiesterases as revealed by molecular simulations. A model system for the enzyme‐substrate complex was prepared on the base of the crystal structure of the EAL domain from the BlrP1 protein complexed with c‐di‐GMP. The nucleophilic hydroxide generated from the bridging water molecule appeared in a favorable position for attack on the phosphorus atom of c‐di‐GMP. The most difficult task was to find a pathway for a proton transfer to the O3' atom of c‐di‐GMP to promote the O3'? P bond cleavage. We show that the hydrogen bond network extended over the chain of water molecules in the enzyme active site and the Glu359 and Asp303 side chains provides the relevant proton wires. The suggested mechanism is consistent with the structural, mutagenesis, and kinetic experimental studies on the EAL domain phosphodiesterases. Proteins 2016; 84:1670–1680. © 2016 Wiley Periodicals, Inc.     

The effect of wire plane tilt and olive wires on proximal tibia fracture fragment stability and fracture site motion

To analyze the effect of the tilt angle relationship between the crossed wire plane and the bone axis on the stiffness of fine wire external fixation, load-deformation behavior was compared across different tilt angles (0 degree, 10 degrees, and 20 degrees) of the plane containing crossed smooth or olive wires under identical conditions of central axial compression, medial compression-bending, posterior compression-bending, posteromedial compression-bending, and torsion. Stiffness values were calculated from the load-deformation and torque-angle curves. A tilt angle of 20 degrees with olive wires provided significantly greater stiffness compared to smooth wires at any angle in any loading condition (p < 0.05). A tilt angle of 20 degrees with olive wires was also significantly more stiff than a tilt angle of 0 degree with olive wires in any loading condition. In torsion, olive wires with 10 degrees and 20 degrees tilt were not significantly different, while in posterior bending olive wires with 10 degrees tilt were significantly stiffer than olive wires with 0 degree or 20 degrees tilt. With smooth wires, tilting the wire plane caused a decrease in stiffness in posterior bending, posteromedial bending, and torsion. Overall, the use of olive wires in conjunction with tilting the wire plane enhances the fixation stiffness for proximal tibia fractures while allowing more options for wire configurations that avoid neurovascular and musculotendinous structures, and wounds.     

Membrane transport through α-heltical bundles. IV. Preliminary model building investigation of helix-helix interactions

Previously we made order of magnitude estimates that suggested the possibility of forming proton wires between facing α-helices joined by knobs-into-holes packing (Dunker, Marvin, Zaleske & Jones, 1976; Dunker & Marvin, 1978). Such structures may be a feature of membrane proteins. Since our original work, another type of packing, called ridges-into-grooves, has been identified as a way of meshing adjoining α-helices. Using precision (CPK) molecular models, we have investigated the possibility of forming proton wires: (1) in order to improve on our previous order of magnitude estimates, and (2) in order to evaluate the different kinds of packing interfaces for their ability to support stereochemically feasible proton wires.We found knobs-into-holes and ridges-into=grooves packing to support exactly the same proton wires. The positions of the side chains are determined more by the geometry of the hydrogen-bonding network rather than by the type of packing originally used to locate the appropriate residues. Thus, we suggest that a new name is needed for such packing, which we propose to call “H-bond packing”.In our earlier investigations we arbitrarily restricted our attention to proton wires parallel to the packing interface. By lifting this restriction, we found it possible to construct many additional types of wires. The model building exercises suggested that both parallel-to-the-interface and non-parallel-to-the-interface wires are feasible, except that the non-parallel wires are restricted in length depending on the angle with the interface, whereas the parallel wires apparently can be continued indefinitely. The various types of wires share local hydrogen bonding patterns and so could easily connect together.In our model building of several representative wires, we investigated the limitations with regard to type of and combinations of side chains. We also determined possible variations in the origins of such side chains on the helical backbones.These preliminary model building studies provide the basis for determining possible hydrogen bonding between the helical segments of membrane proteins. From these data we are formulating preliminary proposals for the helix-helix interactions of the bacteriorhodopsin molecule, which are to be presented in future paper.     

Automatic localization of curved wires used in brachytherapy. The COUREP program.

The localization of sources used for brachytherapy has become more difficult since radium sources are replaced by flexible wires such as Iridium 192 wires. We describe the COUREP program which automatically reconstructs the real 3-D arrangement of curved wires from two orthogonal radiographs of the implant. The data used by the program consist mainly of the projections of the wires on the radiographs expressed as the co-ordinates of points selected on these projections individually on each radiograph. The output consists of the 3-D co-ordinates of points located on the wires with optionally a graphic output including the plotting of a perspective view of the implant.     

Analysis of the electrophoretic migration of DNA frayed wires

We analyzed the electrophoretic behaviour of the unusual multi-stranded DNA complexes, frayed wires, in polyacrylamide gels under non-denaturing conditions. Frayed wires arise from the association of several strands of a parent oligonucleotide that possesses long terminal runs of consecutive guanines. According to the structural model proposed for frayed wires, there are two distinct conformational domains, a guanine stem and single stranded arms displaced from the stem. The presence of the two domains affects the electrophoretic migration of the frayed wires, resulting in a greater retardation compared to that of double stranded DNA of the same molecular weight. The degree of retardation is determined by the relative length of the stem and the arms; the complexes with longer arms display a stronger dependence on the total molecular weight. Reptation plots (mobility x molecular weight vs. molecular weight) were used to study the electrophoretic behaviour of frayed wires that arise from the different parent oligonucleotides. The plots are unique for each type of frayed wire. The characteristic parameter, the position of the maximum of the reptation plot, depends on the type of the frayed wire as well as the total gel concentration. The plots become similar when we replot the mobility data taking into account only the single stranded arms of the frayed wires. The positions of the maximum and the overall shape are very close for the four types of frayed wires studied.     

Tensioning technique and fixator stability of a circular external fixator with threaded fine wires

Hybrid external fixators have become more popular over recent years, and although biomechanical tests show varying results from paper to paper, a consistent finding is shear motion occurring at the fracture/osteotomy site during loading. This can be reduced to some extent by frame configuration, and with the use of olive stop wires, but both of these have limitations. We have investigated the use of threaded fine wires in a circular hybrid fixator. A reliable tensioning method is described which minimises movement at the wire-bone interface during tensioning, and threaded wires are seen to offer a significant improvement in fracture site shear stiffness when compared to smooth fine wires.     

Thermal activation of DNA frayed wire formation

Dynamic light scattering has been used to study the formation of stable multistranded DNA complexes called frayed wires. DNA frayed wires arise from the indefinite self-association of oligonucleotides with long terminal tracks of guanines, e.g. d(A15G15). The complexes are stabilized via guanine-guanine interactions resulting in the formation of a guanine stem. Non-guanine portions of the oligonucleotide are disposed away from the stem and form single stranded arms. The indefinite nature of the self-association of these oligonucleotides leads to a distribution of aggregate molecular weights. The distribution arises from aggregated species that differ from one another by the number of self-associated oligonucleotides. In light-scattering experiments, the autocorrelation functions collected for frayed wires are bimodal. The slow mode, often observed for samples containing DNA and other polyelectrolytes, has been ascribed to the formation of large unspecific aggregates due to electrostatic or other long-range interactions. We attribute the fast mode to the translational diffusion of the polydisperse population in the frayed wire sample. We use the mean of the fast mode to characterize the growth of the frayed wires. Consistent with the gel electrophoresis studies, the aggregation of frayed wires is promoted by the presence of magnesium ions and incubation at high temperature. The rate of aggregate formation increases with temperature, indicating the positive activation energy for the reaction. We propose an energy diagram for the formation/disruption of frayed wires revealing the catalytic-like role of the complementary strand in the denaturation of high molecular weight complexes.     

Laser radiation scattering from the wires and fibers of imploding arrays on the Angara-5-1 facility

A method is developed for measurements of laser radiation scattering by wires and fibers in different types of imploding arrays in the initial stage of plasma production at discharge currents per wire of up to 2 kA for aluminum arrays and up to 8 kA for tungsten arrays. The experiments were carried out on the Angara-5-1 facility at a current density in the wires of 10⁸ A/cm² and current growth rate of ～10¹³ A/s. It is found that the indicatrix of laser radiation reflected from the wires (fibers) in cylindrical and conical arrays is modified at currents of 0.1–10 kA per wire (fiber). The experimental data on the reflection and scattering of laser radiation from wires and fibers are compared with the results of numerical simulations of their electric explosion in vacuum. It is proposed that the change in the reflection indicatrix of laser radiation is caused by the onset of thermal instabilities. The typical size of density and temperature inhomogeneities on the wire surface is in a range of 10–20 μm, which probably results in a transition from specular to diffuse reflection of laser radiation. A simultaneous abrupt (over 2–3 ns) reduction in the reflection intensity from several wires of an array indicates a homogeneous distribution of the discharge current over the irradiated wires. This closes the issue of the quality of the contact between the wires and the electrodes. The obtained experimental information is of considerable importance for the development of numerical codes for simulations of the implosion of wire arrays and the refinement of the wire parameters in the initial stage of plasma production.     

Comparison of mechanical properties of orthodontic nickel-titanium wires]

Two packages each, containing 10 wires per package, of different batches of 25 different types of orthodontic archwires made of super-elastic nickel-titanium alloys measuring 0.41 x 0.56 mm2, were investigated. The wires were characterized by obtaining the following measurements at an ambient temperature of 37 degrees: a three-point bending test with the supporting points spaced 10 mm apart, and determination of the torque/bending angle curves using a pure bending test. The force/deflection curves provided the parameters characterizing the super-elastic unloading plateau: average force, slope and endpoint. From the torque/bending angle curves, the parameters average torque, plateau endpoint and the elasticity parameters were determined. Average force (0.8-4.5 N), endpoint (0.2-0.9 mm) and the slope of the unloading plateau (0.2-2.1 N/mm) of the three-point bending test clearly differed for individual wires. Significant differences were also seen for average torque (1.5-11.5 Nmm), unloading plateau endpoint (2.7-20.0 degrees) and elasticity parameters epsilon 4, E4, E5 and E6 in the pure bending test. Individual batches showed only minor differences. The results permit the conclusion to be drawn that super-elasticity is applicable to only a small portion of the wires examined. Although other wires showed super-elastic behaviour, the unloading plateaus has a force level of up to 6 N, and cannot be recommended for orthodontic application. The super-elastic plateau is often of use only for deflections greater than 1.5 mm. The use of super-elastic archwires made of nickel-titanium alloys makes sense only when the elastic properties of the respective wires are known. This makes the provision by the manufacturer of relevant data on the elastic properties of wires a necessity.     

Comparative Reliability Studies and Analysis of Au,Pd-Coated Cu and Pd-Doped Cu Wire in Microelectronics Packaging

This paper compares and discusses the wearout reliability and analysis of Gold (Au), Palladium (Pd) coated Cu and Pd-doped Cu wires used in fineline Ball Grid Array (BGA) package. Intermetallic compound (IMC) thickness measurement has been carried out to estimate the coefficient of diffusion (D_o) under various aging conditions of different bonding wires. Wire pull and ball bond shear strengths have been analyzed and we found smaller variation in Pd-doped Cu wire compared to Au and Pd-doped Cu wire. Au bonds were identified to have faster IMC formation, compared to slower IMC growth of Cu. The obtained weibull slope, β of three bonding wires are greater than 1.0 and belong to wearout reliability data point. Pd-doped Cu wire exhibits larger time-to-failure and cycles-to-failure in both wearout reliability tests in Highly Accelerated Temperature and Humidity (HAST) and Temperature Cycling (TC) tests. This proves Pd-doped Cu wire has a greater potential and higher reliability margin compared to Au and Pd-coated Cu wires.     

Quantitative detection and biological propagation of scrapie seeding activity in vitro facilitate use of prions as model pathogens for disinfection

Prions are pathogens with an unusually high tolerance to inactivation and constitute a complex challenge to the re-processing of surgical instruments. On the other hand, however, they provide an informative paradigm which has been exploited successfully for the development of novel broad-range disinfectants simultaneously active also against bacteria, viruses and fungi. Here we report on the development of a methodological platform that further facilitates the use of scrapie prions as model pathogens for disinfection. We used specifically adapted serial protein misfolding cyclic amplification (PMCA) for the quantitative detection, on steel wires providing model carriers for decontamination, of 263K scrapie seeding activity converting normal protease-sensitive into abnormal protease-resistant prion protein. Reference steel wires carrying defined amounts of scrapie infectivity were used for assay calibration, while scrapie-contaminated test steel wires were subjected to fifteen different procedures for disinfection that yielded scrapie titre reductions of ≤10(1)- to ≥10(5.5)-fold. As confirmed by titration in hamsters the residual scrapie infectivity on test wires could be reliably deduced for all examined disinfection procedures, from our quantitative seeding activity assay. Furthermore, we found that scrapie seeding activity present in 263K hamster brain homogenate or multiplied by PMCA of scrapie-contaminated steel wires both triggered accumulation of protease-resistant prion protein and was further propagated in a novel cell assay for 263K scrapie prions, i.e., cerebral glial cell cultures from hamsters. The findings from our PMCA- and glial cell culture assays revealed scrapie seeding activity as a biochemically and biologically replicative principle in vitro, with the former being quantitatively linked to prion infectivity detected on steel wires in vivo. When combined, our in vitro assays provide an alternative to titrations of biological scrapie infectivity in animals that substantially facilitates the use of prions as potentially highly indicative test agents in the search for novel broad-range disinfectants.