Superoxide dismutase 2 knockdown leads to defects in locomotor activity, sensitivity to paraquat, and increased cuticle pigmentation in Tribolium castaneum

Citation: Tabunoki, H., Gorman, M. J., Dittmer, N. T., & Kanost, M. R. (2016). Superoxide dismutase 2 knockdown leads to defects in locomotor activity, sensitivity to paraquat, and increased cuticle pigmentation in Tribolium castaneum. Scientific Reports, 6, 8. doi:10.1038/srep29583Insects can rapidly adapt to environmental changes through physiological responses. The red flour beetle Tribolium castaneum is widely used as a model insect species. However, the stress-response system of this species remains unclear. Superoxide dismutase 2 (SOD2) is a crucial antioxidative enzyme that is found in mitochondria. T. castaneum SOD2 (TcSOD2) is composed of 215 amino acids, and has an iron/manganese superoxide dismutase domain. qRT-PCR experiments revealed that TcSOD2 was present through all developmental stages. To evaluate TcSOD2 function in T. castaneum, we performed RNAi and also assessed the phenotype and antioxidative tolerance of the knockdown of TcSOD2 by exposing larvae to paraquat. The administration of paraquat resulted in significantly higher 24-h mortality in TcSOD2 knockdown larval groups than in the control groups. The TcSOD2 knockdown adults moved significantly more slowly, had lower ATP content, and exhibited a different body color from the control groups. We found that TcSOD2 dsRNA treatment in larvae resulted in increased expression of tyrosinase and laccase2 mRNA after 10 days. This is the first report showing that TcSOD2 has an antioxidative function and demonstrates that T. castaneum may use an alternative antioxidative system when the SOD2-based system fails

Superoxide dismutase 2 knockdown leads to defects in locomotor activity, sensitivity to paraquat, and increased cuticle pigmentation in Tribolium castaneum

Citation: Tabunoki, H., Gorman, M. J., Dittmer, N. T., & Kanost, M. R. (2016). Superoxide dismutase 2 knockdown leads to defects in locomotor activity, sensitivity to paraquat, and increased cuticle pigmentation in Tribolium castaneum. Scientific Reports, 6, 8. doi:10.1038/srep29583Insects can rapidly adapt to environmental changes through physiological responses. The red flour beetle Tribolium castaneum is widely used as a model insect species. However, the stress-response system of this species remains unclear. Superoxide dismutase 2 (SOD2) is a crucial antioxidative enzyme that is found in mitochondria. T. castaneum SOD2 (TcSOD2) is composed of 215 amino acids, and has an iron/manganese superoxide dismutase domain. qRT-PCR experiments revealed that TcSOD2 was present through all developmental stages. To evaluate TcSOD2 function in T. castaneum, we performed RNAi and also assessed the phenotype and antioxidative tolerance of the knockdown of TcSOD2 by exposing larvae to paraquat. The administration of paraquat resulted in significantly higher 24-h mortality in TcSOD2 knockdown larval groups than in the control groups. The TcSOD2 knockdown adults moved significantly more slowly, had lower ATP content, and exhibited a different body color from the control groups. We found that TcSOD2 dsRNA treatment in larvae resulted in increased expression of tyrosinase and laccase2 mRNA after 10 days. This is the first report showing that TcSOD2 has an antioxidative function and demonstrates that T. castaneum may use an alternative antioxidative system when the SOD2-based system fails

Review

The chalcogen elements oxygen, sulfur, and selenium are essential constituents of side chain functions of natural amino acids. Conversely, no structural and biological function has been discovered so far for the heavier and more metallic tellurium element. In the methionine series, only the sulfur-containing methionine is a proteinogenic amino acid, while selenomethionine and telluromethionine are natural amino acids that are incorporated into proteins most probably because of the tolerance of the methionyl-tRNA synthetase; so far, methoxinine the oxygen analogue has not been discovered in natural compounds. Similarly, the chalcogen analogues of tryptophan and phenylalanine in which the benzene ring has been replaced by the largely isosteric thiophene, selenophene, and more recently, even tellurophene are fully synthetic mimics that are incorporated with more or less efficiency into proteins via the related tryptophanyl- and phenylalanyl-tRNA synthetases, respectively. In the serine/cysteine series, also selenocysteine is a proteinogenic amino acid that is inserted into proteins by a special translation mechanism, while the tellurocysteine is again most probably incorporated into proteins by the tolerance of the cysteinyl-tRNA synthetase. For research purposes, all of these natural and synthetic chalcogen amino acids have been extensively applied in peptide and protein research to exploit their different physicochemical properties for modulating structural and functional properties in synthetic peptides and rDNA expressed proteins as discussed in the following review

Timing molecular motion and production with a synthetic transcriptional clock

The realization of artificial biochemical reaction networks with unique functionality is one of the main challenges for the development of synthetic biology. Due to the reduced number of components, biochemical circuits constructed in vitro promise to be more amenable to systematic design and quantitative assessment than circuits embedded within living organisms. To make good on that promise, effective methods for composing subsystems into larger systems are needed. Here we used an artificial biochemical oscillator based on in vitro transcription and RNA degradation reactions to drive a variety of “load” processes such as the operation of a DNA-based nanomechanical device (“DNA tweezers”) or the production of a functional RNA molecule (an aptamer for malachite green). We implemented several mechanisms for coupling the load processes to the oscillator circuit and compared them based on how much the load affected the frequency and amplitude of the core oscillator, and how much of the load was effectively driven. Based on heuristic insights and computational modeling, an “insulator circuit” was developed, which strongly reduced the detrimental influence of the load on the oscillator circuit. Understanding how to design effective insulation between biochemical subsystems will be critical for the synthesis of larger and more complex systems

Cuticular protein with a low complexity sequence becomes cross-linked during insect cuticle sclerotization and is required for the adult molt

Citation: Mun, S., Noh, M. Y., Dittmer, N. T., Muthukrishnan, S., Kramer, K. J., Kanost, M. R., & Arakane, Y. (2015). Cuticular protein with a low complexity sequence becomes cross-linked during insect cuticle sclerotization and is required for the adult molt. Scientific Reports, 5, 11. doi:10.1038/srep10484In the insect cuticle, structural proteins (CPs) and the polysaccharide chitin are the major components. It has been hypothesized that CPs are cross-linked to other CPs and possibly to chitin by quinones or quinone methides produced by the laccase2-mediated oxidation of N-acylcatechols. In this study we investigated functions of TcCP30, the third most abundant CP in protein extracts of elytra (wing covers) from Tribolium castaneum adults. The mature TcCP30 protein has a low complexity and highly polar amino acid sequence. TcCP30 is localized with chitin in horizontal laminae and vertically oriented columnar structures in rigid cuticles, but not in soft and membranous cuticles. Immunoblot analysis revealed that TcCP30 undergoes laccase2-mediated cross-linking during cuticle maturation in vivo, a process confirmed in vitro using recombinant rTcCP30. We identified TcCPR27 and TcCPR18, the two most abundant proteins in the elytra, as putative crosslinking partners of TcCP30. RNAi for the TcCP30 gene had no effect on larval and pupal growth and development. However, during adult eclosion, similar to 70% of the adults were unable to shed their exuvium and died. These results support the hypothesis that TcCP30 plays an integral role as a cross-linked structural protein in the formation of lightweight rigid cuticle of the beetle

Lead-free high-temperature dielectrics with wide operational range

The dielectric, electrical and structural properties of (1-x) (0.94Bi(1/2)Na(1/2)TiO(3)-0.06BaTiO(3))-xK(0.5)Na(0.5)NbO(3) (BNT-BT-xKNN) with x=0.09, 0.12, 0.15, and 0.18 were investigated as potential candidates for high-temperature capacitors with a working temperature far beyond 200 degrees C. Temperature dependent dielectric permittivity (epsilon) showed two local broad maxima that at the optimal composition of KNN (x=0.18) are combined to form a plateau. This then results in a highly temperature-insensitive permittivity up to similar to 300 degrees C at the expense of a small reduction in absolute permittivity values. High-temperature in situ x-ray diffraction study showed pseudocubic symmetry without obvious structural changes, which implies that the dielectric anomalies observed could only be a consequence of a slight change in space group. BNT-BT-0.18KNN showed a permittivity of similar to 2150 at the frequency of 1 kHz at 150 degrees C with a normalized permittivity epsilon/epsilon(150 degrees C) varying no more than +/- 10% from 43 to 319 degrees C. With very good electrical properties persisting up to 300 degrees C, i.e., a resistivity on the order of magnitude of 10(8) Omega m and the RC constant of about 1 s, the examined BNT-BT-xKNN compositions present a good starting point for the development of high-temperature capacitor materials.open343

Code generation based on inference and controlled natural language input

Over time the level of abstraction embodied in programming languages has continued to grow. Paradoxically, most programming languages still require programmers to conform to the language\u27s rigid constructs. These constructs have been implemented in the name of efficiency for the computer. However, the continual increase in computing power allows us to consider techniques not so limited. To this end, we have created CABERNET, a Controlled Natural Language (CNL) based approach to program creation. CABERNET allows programmers to use a simple outline-based syntax. This syntax enables increased programmer efficiency. CNLs have previously been used to document requirements. We have taken this approach beyond the typical application of creating requirements documents to creating functional programs. Using heuristics and inference to analyze and determine the programmer\u27s intent, the CABERNET toolchain can create functional mobile applications. This approach allows programs to align with how humans think rather than how computers process information. Using customizable templates, a CABERNET application can be processed to run on multiple run-time environments. Since processing a CABERNET program file results in a native application program, performance is maintained. This research explores whether a CNL-based programming tool can provide a readable, flexible, extensible, and easy-to-learn development methodology. To answer this question, we compared sample applications created in Swift, SwiftUI, and a prototype of the CABERNET toolchain. The CABERNET implementations were consistently shorter than those produced in the other two languages. In addition, users surveyed consistently found the CABERNET samples easier to understand

Local structure change evidenced by temperature-dependent elastic measurements: Case study on Bi1/2Na1/2TiO3-based lead-free relaxor piezoceramics

The temperature-dependent Young's modulus Y(T) of the lead-free piezoceramics of 0.8Bi(1/2)Na(1/2)TiO(3)-0.2Bi(1/2)K(1/2)TiO(3) (20BKT) and 0.96(0.8Bi(1/2)Na(1/2)TiO(3)-0.2Bi(1/2)K(1/2)TiO(3))-0.04 BiZn1/2Ti1/2O3 (4BZT) is measured with the impulse excitation technique and contrasted with corresponding dielectric and structural data. While the dielectric properties suggest a phase transition, the high resolution XRD patterns remain virtually unchanged from room temperature up to high temperatures, confirming no change in their long-range order. In contrast, the elastic properties indicate a broad and diffuse ferroelastic transition denoted by a minimum in Y(T). By analogy to the elastic and dielectric data of PbZrxTi1-xO3 and PLZT, it is concluded that 20BKT and 4BZT are relaxors with polar nanoregions embedded in a metrically cubic matrix. Interestingly, no indication for the freezing temperature was reflected in any of the employed measurement techniques. From the saturation of Y(T), it is suggested that the Burns temperature may be approximated as 700 degrees C. Moreover, it is found that the modification with the ternary end-member BiZn1/2Ti1/2O3 results in an increase in Young's modulus. A comparison with the Bi1/2Na1/2TiO3-BaTiO3-K0.5Na0.5NbO3 yields the same results.open0

Investigation of the depolarisation transition in Bi-based relaxor ferroelectrics

The loss of macroscopic polarisation in relaxor ferroelectric (Na0.8K0.2)(1/2)Bi1/2TiO3 ceramics doped with BiZn1/2Ti1/2O3 has been studied by electrical and structural methods. These indicate that the phenomena that are coupled in a displacive phase transition are not necessarily coupled in the depolarisation of Na1/2Bi1/2TiO3-based relaxors and a concept of correlated and uncorrelated switching of dipoles within adjacent unit cells is used to explain this. Second harmonic generation performed on poled ceramics during heating yields values of the freezing temperature and shows a broad temperature range of similar to 100 degrees C across which the structure changes from field-induced ferroelectric to an equilibrium-state ergodic relaxor. Electrical poling at room temperature causes poled regions to increase in size by similar to 2 orders of magnitude. A model illustrating the main steps in thermal depolarisation is described that does not require a phase transition to take place on a unit cell level.open1