Broad-specificity immunoassays for sulfonamide detection: immunochemical strategy for generic antibodies and competitors

Development of antibodies with broad specificity recognition for sulfonamide drugs was found to be surprisingly difficult when conventional immunochemical strategies were applied to hapten design. To improve the cross-reactivity pattern of antibodies for the family of sulfonamide drugs, a novel strategy based on the single-ring (fragment-derived) hapten moieties with different spacer substituent lengths was employed for the preparation of immunogens, coating conjugates, and enzyme competitors. The rabbit antibodies raised against a common (one-ring) p-aminobenzenesulfonamide hapten moiety (attached to a carrier protein through the N-1 position) in combination with a homologous hapten-peroxidase tracer allowed the detection of 15 sulfonamide species at the maximum residue limit level using direct ELISA. The two-ring 6-(4-aminobenzensulfonylamino)hexanoic hapten mimics, previously reported in the literature as a weak generic antigen, generated surprisingly superior immune responses in rabbits. The antibodies raised against this two-ring hapten were capable of detecting at least 19 and 17 sulfonamides in a direct ELISA system at the regulatory level with sensitivities corresponding to 20 and 50% binding inhibition, respectively. A negligible cross-reaction with N4 metabolites makes it possible to measure responses of parent sulfonamides in the presence of their metabolized forms. In skimmed milk, the highest limit of detection (LOD) for sulfacetamide defined as 20% inhibition was 65.2 microg x L(-1) (IC20 value), whereas the additional 18 sulfonamides tested exhibited LODs in the range of 0.2-36.8 microg x L(-1). This sensitivity allows simple multisulfonamide tests to be established for use in the laboratory or on site.     

On the Enzymatic Formation of Metal Base Pairs with Thiolated and pKa-Perturbed Nucleotides

The formation of artificial metal base pairs is an alluring and versatile method for the functionalization of nucleic acids. Access to DNA functionalized with metal base pairs is granted mainly by solid-phase synthesis. An alternative, yet underexplored method, envisions the installation of metal base pairs through the polymerization of modified nucleoside triphosphates. Herein, we have explored the possibility of using thiolated and pK_a-perturbed nucleotides for the enzymatic construction of artificial metal base pairs. The thiolated nucleotides S2C, S6G, and S4T as well as the fluorinated analogue 5FU are readily incorporated opposite a templating S4T nucleotide through the guidance of metal cations. Multiple incorporation of the modified nucleotides along with polymerase bypass of the unnatural base pairs are also possible under certain conditions. The thiolated nucleotides S4T, S4T, S2C, and S6G were also shown to be compatible with the synthesis of modified, high molecular weight single-stranded (ss)DNA products through TdT-mediated tailing reactions. Thus, sulfur-substitution and pK_a perturbation represent alternative strategies for the design of modified nucleotides compatible with the enzymatic construction of metal base pairs.     

Switching between negative and positive electrorheological effect of g-C3N4 by copper ions doping

Investigation of pristine g-C₃N₄ and copper ions doped g-C₃N₄ as dispersed phases in silicone-oil based electrorheological (ER) fluids is reported for the first time. Pristine g-C₃N₄ was prepared via standard thermal polycondensation of urea. Introduction of Cu ions into g-C₃N₄ polymeric network was performed by treatment of pristine powder by a solution containing tetraammoniumdiaquacopper(II) complex ions followed by annealing. The structure and properties of products were revealed with the aid of XRD, FTIR, UV–Vis and XPS, and complemented by SEM investigation of morphology, pycnometry, BET analysis, and conductivity measurements. The response of prepared ER fluids to an external electric field was examined by rheometry and the effects visualized with optical microscopy. While ER fluids based on g-C₃N₄ exhibited negative ER effect (decrease in viscosity when the field is switched-on), ER fluids based on a g-C₃N₄/Cu-doped analogue exhibited positive ER effect. Dielectric spectroscopy using Havriliak-Negami model revealed that introduction of the dopant almost doubled the dielectric relaxation strength while the relaxation time was halved. The ability of g-C₃N₄/Cu as a candidate for further studies necessary to increase the ER effect was demonstrated. Thereto, described modification of g-C₃N₄ by copper ions shall be applicable also for other metals forming ammonia complexes.     

Digitally reconstructing Van Gogh's Field with Irises near Arles part 3: Determining the original colors

In earlier articles, we determined the spatial distributions and concentrations of all pigments used by Van Gogh in his painting Field with Irises near Arles. The colors of some pigments are expected to have changed over time, especially those of chrome yellow, cochineal, and eosin lake. For all pigments in this painting, we made physical paint reconstructions by following historical sources on raw materials and production processes, and we determined their optical properties. We combined this with pigment concentration maps to reconstruct the original colors of the painting digitally. When substituting the reconstruction paints into the calculations, we found that technical‐scientific data was not sufficient to resolve several issues. In those cases, discussions within the broad interdisciplinary team allowed us to make informed decisions. These issues refer to the representation of the sky area, and the original contributions of the red lake pigments to local colors. The digitally reconstructed colors of the painting show that due to discoloring of red lake pigments, the irises in the field have changed from a warm purple to purplish blue, and many pink spots in the field have turned to white. The range of yellows in the field has decreased and partly turned to dark brown. The digital reconstruction gives a better understanding of the color scheme used by Van Gogh when compared to remarks the artist made in letters when describing this painting. Also, the original color composition is seen to be aligned with color theories on which Van Gogh based his work.     

An approach to torque and temperature thread by thread on tapping

During internal threading, small alterations in cutting parameters, tool geometry, or process characteristics produce considerable effects on torque and te     

Towards (dis)continuity of agricultural wetlands: Latvia’s polder landscapes after Soviet productivism

The concepts of agricultural regimes in advanced economies, such as productivism or non/neo/post-productivism, have been critically debated over the last decades to understand the transition and diversity of modern agriculture. We explore these concepts to understand the environmentally vulnerable landscape of agricultural wetlands in Latvia that, during the era of Soviet high modernism (productivist agricultural regime), have been converted into polders as part of a mass drainage movement. Today, these post-Soviet agro-polders can be characterised as antipodes in relation to integrity of heritage, ecology and the socio-economics of agricultural concerns. Building on case studies, wider political contexts and current debates on agricultural transitions, the paper traces the transition of wetlands through agricultural regimes and unfolds the various pathways for current polder landscapes. This paper concludes with critical notes on the contemporary co-existence of different agro-polder use of which the most incoherent are the non-productivist actions producing landscapes of mere agricultural appearance.     

Components for high speed atomic force microscopy

Many applications in materials science, life science and process control would benefit from atomic force microscopes (AFM) with higher scan speeds. To achieve this, the performance of many of the AFM components has to be increased. In this work, we focus on the cantilever sensor, the scanning unit and the data acquisition. We manufactured 10 microm wide cantilevers which combine high resonance frequencies with low spring constants (160-360 kHz with spring constants of 1-5 pN/nm). For the scanning unit, we developed a new scanner principle, based on stack piezos, which allows the construction of a scanner with 15 microm scan range while retaining high resonance frequencies (>10 kHz). To drive the AFM at high scan speeds and record the height and error signal, we implemented a fast Data Acquisition (DAQ) system based on a commercial DAQ card and a LabView user interface capable of recording 30 frames per second at 150 x 150 pixels.     

On total internal reflection investigation of nanoparticles by integrated micro-fluidic system

We report on a novel sensor for characterization of nanoparticles colloidal suspensions. We employ a diffraction grating under total internal reflection for investigation of nanodisperse fluids passing through an integrated microfluidic channel. Dispersions containing polymeric, metallic, and ferromagnetic nanoparticles are studied. Using this device, we can accurately determine in real-time the specific refractive index for the nanoparticle suspension and the nanoparticle concentration. The nanoparticle concentrations can be calculated with a resolution of 0.3-0.5 wt% for polymeric nanoparticles, 0.03-0.05 wt% for metallic nanoparticles, and 0.05-0.1 wt% for ferromagnetic nanoparticles. This translates to an effective refractive index that can be determined with an accuracy of 7 x 10(-4) for the polymeric and 2 x 10(-4) for the metallic and ferromagnetic dispersions.