Duplex Structure Research Articles

Design of a Duplex-to-Complex Structure-Switching Approach for the Homogeneous Determination of Marine Biotoxins in Water

In this report, we describe a fluorescent assay for the detection of six marine toxins in water. The mechanism of detection is based on a duplex-to-complex structure-switching approach. The six aptamers specific to the targeted cyanotoxins were conjugated to a fluorescent dye, carboxyfluorescein (FAM). In parallel, complementary DNA (cDNA) sequences specific to each aptamer were conjugated to a fluorescence quencher BHQ1. In the absence of the target, an aptamer–cDNA duplex structure is formed, and the fluorescence is quenched. By adding the toxin, the aptamer tends to bind to its target and releases the cDNA. The fluorescence intensity is consequently restored after the formation of the complex aptamer–toxin, where the fluorescence recovery is directly correlated with the analyte concentration. Based on this principle, a highly sensitive detection of the six marine toxins was achieved, with the limits of detection of 0.15, 0.06, 0.075, 0.027, 0.041, and 0.026 nM for microcystin-LR, anatoxin-α, saxitoxin, cylindrospermopsin, okadaic acid, and brevetoxin, respectively. Moreover, each aptameric assay showed a very good selectivity towards the other five marine toxins. Finally, the developed technique was applied for the detection of the six toxins in spiked water samples with excellent recoveries.

Recognition-Encoded Molecules: A Minimal Self-Replicator.

Nucleic acids, with their unique duplex structure, which is key for information replication, have sparked interest in self-replication's role in life's origins. Early template-based replicators, initially built on short oligonucleotides, expanded to include peptides and synthetic molecules. We explore here the potential of a class of synthetic duplex-forming oligoanilines, as self-replicators. We have recently developed oligoanilines equipped with 2-trifluoromethylphenol-phosphine oxide H-bond base pairs and we investigate whether the imine formed between aniline and aldehyde complementary monomers can self-replicate. Despite lacking a clear sigmoidal kinetic profile, control experiments with a methylated donor and a competitive inhibitor support self-replication. Further investigations with the reduced aniline dimer demonstrate templated synthesis, revealing a characteristic parabolic growth. After showing sequence selective duplex formation, templated synthesis and the emergence of catalytic function, the self-replication behaviour further suggests that the unique properties of nucleic acids can be paralleled by synthetic recognition-encoded molecules.

Split G-Quadruplex Programmed Recyclable AIE-Biosensor for Label-Free Detection of miRNA in Acute Kidney Injury.

We herein rationally designed a target-recyclable AIE-biosensor based on a split G-quadruplex for label-free detection of miRNA in acute kidney injury. Initially, the PG was in an "OFF" state, and the two split segments (G4-a and G4-b) of G4 were tethered at the two terminals of P1 and far away from each other due to the rigid duplex structure formed by the partially complementary intermediate sequences of P2 and P1, bringing MG with quenched fluorescence. In the presence of target, the 5'-PO4 P2 was displaced from PG probe and competitively hybridized with target, which led to G4-a and G4-b tending to form an intact intermolecular G-quadruplex, providing sites for MG intercalation, thus generating an activated "ON" fluorescence signal due to the restriction of intermolecular motion. Successively, relying on the λ-exonuclease (λ-Exo) cleavage reaction-assisted target recycling, more amounts of targets will be liberated, accompanied by forming more G-quadruplex and binding more MG, resulting in a strong fluorescence signal, further realizing the sensitive detection of the targets. As a proof of concept, miRNA-21 was chosen as the model target. Endowing with the precise target recognition and efficient cleavage activity of λ-Exo, the AIE-biosensor exhibited excellent detection sensitivity and specificity, which could quantitatively detect miRNA-21 down to 10.36 fM with a single mismatch specificity. The results revealed that the distinctive attributes of noninvasive, simple, and efficient in this G-quadruplex-based AIE-biosensor offered promising prospects for extensive applications in AKI screening and early clinical diagnosis.

Preparation of bainite phase microstructure by laser-electromagnetic induction hybrid solid-state phase transformation

Increasing the proportion of bainite and martensite phases is one of the effective ways to enhance the toughness of medium-carbon low-alloy steel(42CrMo). In this paper, the surface heat treatment of 42CrMo steel is carried out by using a laser heat source to achieve complete austenitization, then with electromagnetic induction heating for several times to implement insulation, and eventually to promote bainite phase transformation. In conjunction with the temperature history curves, OM, SEM, and EBSD are used to characterize the bainite morphology and percentage of bainite via different parameters of the hybrid process. The results demonstrate that the hybrid process combined with twice electromagnetic induction heating can yield a considerable proportion of bainite phase in 42CrMo steel. The resulting duplex structure, consisting of bainite and a minor fraction of martensite, exhibits a significant increase (81.1%) in tensile strength, while maintaining similar impact toughness. The elongation after fracture only has a slight reduction of 5.5% compared to that of the substrate. This innovative process provides a practical alternative to traditional bainite isothermal treatment method which must perform in a heat treatment furnace. Therefore, it is presented as a promising solution for enhancing the mechanical properties of medium-carbon low-alloy high strength steels.

Structural architecture and metamorphism of the Mayombe Chain and Niari Basin (West Congo Belt) in Congo Brazzaville

Geological mapping of the Mayombe Chain and Niari Basin of Congo Brazzaville allows for the first time defining the structural architecture and metamorphism of the West Congo Belt. Four different tectono-metamorphic domains, separated by crustal-scale shear zones, are now distinguished (Niari Basin (NB), Eastern (EMC), Central (CMC) and Western (WMC) Mayombe Chain).The NB is marked by only weak regional deformation under middle to upper diagenetic conditions. It is delimited in the west from the EMC by the Mount Belo Shear Zone forming the terminal thrust system of the West Congo orogen.The tectonic style in the EMC is characterized by discrete, widely-spaced low-angle thrusts, reverse faults and strike-slip faults resulting in the formation of duplex and/or positive flower structures. Off these high-strain zones, the rocks are folded into gentle syn- and anticlines. Penetrative schistosity starts in shales in the western part. The metamorphism increases from eastern anchizonal conditions to lower greenschist facies in the west. The EMC is juxtaposed along the Moukondo thrust/back-thrust system with the CMC.The CMC is typified by open to closed upright to NE-verging folds, S1 schistosity with moderate to steep SW dips, onset of regional crenulation cleavage (S2), frequent reverse thrusts and numerous faults. Metamorphic conditions remain in the greenschist facies. The Loukenéné-Mandji Thrust marks the CMC-WMC contact and coincides with a jump in metamorphic grade marked by biotite-in.The WMC consists of Palaeoproterozoic basement stacked with Neoproterozoic rocks. Autochthonous Palaeoproterozoic gneiss and schist record Late Eburnean sedimentation, magmatism and metamorphism between 2110 and 1970 Ma, which are compared with the Eburnean history in Gabon and the Transamazonian orogeny in Brazil. The allochthonous Bikossi Group was thrust during the Pan-African event from the west over Tonian metavolcaniclastic and plutonic rocks before further folding and stacking of both units. The intensity of Pan-African deformation increases from open to closed folds with spaced cleavage in the southeast of the WMC to thrust-dominated tectonics in the northwest, where the Palaeo- and Neoproterozoic rocks are transposed into parallelism with the pronounced schistosity.Geochronology of illite and muscovite documents two Pan-African events at 590-570 Ma (M1) and at 520-500 Ma (M2) that are related to the main collisional and late thermal events in the Araçuai-Ribeira Belt in Brazil. Metamorphic isogrades shifted from M1 to M2 for more than 30 km to the west. Detrital mica and metamorphic illite of the Mpioka Group record M1 and M2, respectively constraining sedimentary deposition between 570 and 520 Ma, which implies the interpretation of the group as molasse of the West Congo Belt.

Tectonic evolution of the eastern margin of the Northern South Yellow Sea Basin in the Yellow Sea since the Late Cretaceous

The South Yellow Sea Basin is the largest sedimentary basin in the Yellow Sea. The Gunsan Basin in the central eastern part of the Northern South Yellow Sea Basin comprises the Western, Central, and Eastern Subbasins. The Eastern Subbasin marks the eastern margin of the South Yellow Sea Basin. Interpretation of multi-channel seismic profiles and balanced cross-section restoration of depth-converted seismic profiles reveal the structural characteristics and evolution of the Eastern Subbasin. The subbasin comprises three groups of faults: NW-SE, NE-SW, and NNE-SSW trending faults. The subsidence pattern of the subbasin, derived from the cross-section restoration analysis, indicates five distinguished evolution phases: (i) rapid subsidence in the Late Cretaceous, (ii) slow subsidence from the Paleocene to the Eocene, (iii) accelerated subsidence in the Oligocene, (iv) alternation of the uplift and subsidence in the Early Miocene, and (v) gradual subsidence since the Middle Miocene. The main and moderate subsidence can be explained by the combination of extension in the SE and NE-SW directions that formed double duplex structures. We suggest that the NW oblique subduction of the Pacific Plate under the NE Asia margin induced both extension toward the trench and dextral strike-slip parallel to the margin. The extension toward the trench caused SE transtension in a local pull-apart setting, whereas the dextral strike-slip parallel to the margin caused NE-SW extension, inducing more significant subsidence. The combined processes resulted in the progressive development of nested duplex structures. The evolution of the Eastern Subbasin is not compatible with previously suggested models for the western part of the South Yellow Sea Basin including foreland basin formation and transtension induced by branch faults of the Tan-Lu Fault.

Enhanced Recognition of a Herbal Compound Epiberberine by a DNA Quadruplex-Duplex Structure.

The small molecule epiberberine (EPI) is a natural alkaloid with versatile bioactivities against several diseases including cancer and bacterial infection. EPI can induce the formation of a unique binding pocket at the 5' side of a human telomeric G-quadruplex (HTG) sequence with four telomeric repeats (Q4), resulting in a nanomolar binding affinity (KD approximately 26 nM) with significant fluorescence enhancement upon binding. It is important to understand (1) how EPI binding affects HTG structural stability and (2) how enhanced EPI binding may be achieved through the engineering of the DNA binding pocket. In this work, the EPI-binding-induced HTG structure stabilization effect was probed by a peptide nucleic acid (PNA) invasion assay in combination with a series of biophysical techniques. We show that the PNA invasion-based method may be useful for the characterization of compounds binding to DNA (and RNA) structures under physiological conditions without the need to vary the solution temperature or buffer components, which are typically needed for structural stability characterization. Importantly, the combination of theoretical modeling and experimental quantification allows us to successfully engineer Q4 derivative Q4-ds-A by a simple extension of a duplex structure to Q4 at the 5' end. Q4-ds-A is an excellent EPI binder with a KD of 8 nM, with the binding enhancement achieved through the preformation of a binding pocket and a reduced dissociation rate. The tight binding of Q4 and Q4-ds-A with EPI allows us to develop a novel magnetic bead-based affinity purification system to effectively extract EPI from Rhizoma coptidis (Huang Lian) extracts.

Ultrasensitive Electrochemiluminescence Biosensor Based on DNA-Bio-Bar-Code and Hybridization Chain Reaction Dual Signal Amplification for Exosomes Detection.

Exosomes have received considerable attention as potent reference markers for the diagnosis of various neoplasms due to their close and direct relationship with the proliferation, adhesion, and migration of tumor. The ultrasensitive detection of cancer-derived low-abundance exosomes is imperative, but still a great challenge. Herein, we report an electrochemiluminescence (ECL) biosensor based on the DNA-bio-bar-code and hybridization chain reaction (HCR)-mediated dual signal amplification for the ultrasensitive detection of cancer-derived exosomes. In this system, two types of aptamers were modified on the magnetic nanoprobe (MNPs) and gold nanoparticles (AuNPs) with numerous bio-bar-code DNA, respectively, which formed "sandwich" structures in the presence of specific target exosomes. The "sandwich" structures were separated under magnetic field, and the numerous bio-bar-code DNA were released by dissolving AuNPs. The released bio-bar-code DNA triggered the HCR procedure to produce a good deal of long DNA duplex structure for embedding in hemin, which generated strong ECL signal in the presence of coreactors for ultrasensitive detection of exosomes. Under the optimal conditions, it exhibited a good linearly of exosomes ranging from 10 to 104 exosomes particle μL-1 with limit of detection down to 5.01 exosome particle μL-1. Furthermore, the high ratio of ECL signal and minor change of ECL intensity indicated the good specificity, stability, and repeatability of this ECL biosensor. Given the good performance for exosome analysis, this ultrasensitive ECL biosensor has a promising application in the clinical diagnosis of early cancers.

Phase evolution, mechanical properties, and corrosion resistance of Ti2NbVAl0.3Zrx lightweight refractory high-entropy alloys

Herein, a series of Ti2NbVAl0.3Zrx (x = 0.25, 0.5, 0.75, 1) light refractory high entropy alloys were designed and prepared based on the concept of B2 ordering and solid solution strengthening. The Zr0.25, Zr0.5, and Zr0.75 alloys exhibited a disordered BCC + ordered B2 duplex structure, while the Zr1 alloy displayed a BCC single-phase structure. The addition of Zr hinders the formation of B2 ordering, transforming the alloy from a BCC + B2 duplex structure to a BCC single-phase structure. The Zr0.75 alloy has a yield strength of 1022 MPa and a tensile strain of ∼9.4 %. Furthermore, electrochemical tests were conducted in 3.5 wt% NaCl solution revealed that the Ti2NbVAl0.3Zrx LRHEAs showed high resistance to pitting corrosion. Incorporating the Zr element enhanced the polarization resistance and reduced the corrosion rate; however, excessive Zr content led to a shift in micro-galvanic corrosion from the nanoscale to the microscale, resulting in pitting corrosion. The passivation film compositions of the four alloys formed at a potential of 0.7 VSCE were analyzed by X-ray photoelectron spectroscopy, and their corrosion resistance in a chlorine-containing environment was examined in depth.

Microstructure, thermal stability and soft magnetic properties of Fe81B10P8Cu1 amorphous/nanocrystalline alloy refined by slags with low basicity

The amorphous/nanocrystalline duplex structure is stable and exhibits excellent soft magnetic properties. However, the presence of impurities inevitably impacts the performance of the products. In this work, the impact of refining slags with various basicities (R=1, 0.66, 0.5, 0.4) on the microstructure, thermal stability, and soft magnetic properties of Fe81B10P8Cu1 alloy was investigated by varying the composition ratio x:1:1 (x = 2, 3, 4, & 5) of B2O3:MnO:MgO. The utilization of refining slag with low basicity is beneficial for removing impurity, enhancing magnetic exchange interactions and reducing magnetic anisotropy in amorphous alloy, thereby improving its soft magnetic properties. Notably, when the refining slag composition of B2O3:MnO:MgO is 4:1:1, the Fe81B10P8Cu1 nanocrystalline alloy exhibits exceptional soft magnetic properties and the smallest grain size, with a Bs of 1.717 T, Hc of 3.85 A/m and an average grain size of 23.84 nm. These findings are significant for enhancing the performance of Fe-based nanocrystalline alloys.

Accelerated Screening of Alternative DNA Base‐Organic Molecule‐Base Architectures via Integrated Theory and Experiment

AbstractOrganic molecule‐mediated noncanonical DNA self‐assembly expands the standard DNA base‐pairing alphabets. However, only a very limited number of small molecules have been recognized as mediators because of the tedious and complicated experiments like crystallization and microscopy imaging. Here we present an integrative screening protocol incorporating molecular dynamics (MD) for fast theoretical simulation and native polyacrylamide gel electrophoresis for convenient experimental validation. Melamine, the molecule that was confirmed mediating noncanonical DNA base‐pairing, and 38 other candidate molecules were applied to demonstrate the feasibility of this protocol. We successfully identified seven stable noncanonical DNA duplex structures, and another eight novel structures with sub‐stability. In addition, we discovered that hairpins at both ends can significantly stabilize the noncanonical DNA structures, providing a guideline to design small organic molecule‐incorporated DNA structures. Such an efficient screening protocol will accelerate the design of alternative DNA‐molecule architectures beyond Watson–Crick pairs. Considering the wide range of potential mediators, it will also facilitate applications such as noncovalent, highly dense loading of drug molecules in DNA‐based delivery system and probe design for sensitive detection of certain molecules.

Accelerated Screening of Alternative DNA Base-Organic Molecule-Base Architectures via Integrated Theory and Experiment.

Organic molecule-mediated noncanonical DNA self-assembly expands the standard DNA base-pairing alphabets. However, only a very limited number of small molecules have been recognized as mediators because of the tedious and complicated experiments like crystallization and microscopy imaging. Here we present an integrative screening protocol incorporating molecular dynamics (MD) for fast theoretical simulation and native polyacrylamide gel electrophoresis for convenient experimental validation. Melamine, the molecule that was confirmed mediating noncanonical DNA base-pairing, and 38 other candidate molecules were applied to demonstrate the feasibility of this protocol. We successfully identified seven stable noncanonical DNA duplex structures, and another eight novel structures with sub-stability. In addition, we discovered that hairpins at both ends can significantly stabilize the noncanonical DNA structures, providing a guideline to design small organic molecule-incorporated DNA structures. Such an efficient screening protocol will accelerate the design of alternative DNA-molecule architectures beyond Watson-Crick pairs. Considering the wide range of potential mediators, it will also facilitate applications such as noncovalent, highly dense loading of drug molecules in DNA-based delivery system and probe design for sensitive detection of certain molecules.

Reversible Control of Gene Expression by Guest-Modified Adenosines in a Cell-Free System via Host-Guest Interaction.

Gene expression technology has become an indispensable tool for elucidating biological processes and developing biotechnology. Cell-free gene expression (CFE) systems offer a fundamental platform for gene expression-based technology, in which the reversible and programmable control of transcription can expand its use in synthetic biology and medicine. This study shows that CFE can be controlled via the host-guest interaction of cucurbit[7]uril (CB[7]) with N6-guest-modified adenosines. These adenosine derivatives were conveniently incorporated into the DNA strand using a post-synthetic approach and formed a selective and stable base pair with complementary thymidine in DNA. Meanwhile, alternate addition of CB[7] and the exchanging guest molecule induced the reversible formation of a duplex structure through the formation and dissociation of a bulky complex on DNA. The kinetics of the reversibility was fine-tuned by changing the size of the modified guest moieties. When incorporated into a specific region of the T7 promoter sequence, the guest-modified adenosines enabled tight and reversible control of in vitro transcription and protein expression in the CFE system. This study marks the first utility of the host-guest interaction for gene expression control in the CFE system, opening new avenues for developing DNA-based technology, particularly for precise gene therapy and DNA nanotechnology.

Target-promoted autocatalytic hairpin assembly of bivalent DNAzymes for sensitive and label-free electrochemical metallothionein assay

Metallothionein (MT) has shown to be an important biomarker for environmental monitoring and various diseases, due to its significant binding ability to heavy metal ions. On the basis of such a characteristic and the Hg2+-stabilized DNA duplex (Hg2+-dsDNA) probe, as well as a new autocatalytic hairpin assembly (aCHA)/DNAzyme cascaded signal enhancement strategy, the construction of a highly sensitive and label-free electrochemical MT biosensor is described. Target MT molecules bind Hg2+ in Hg2+-dsDNA to disrupt the duplex structure and to release ssDNA sequences, which trigger subsequent aCHA for efficient production of mimic aCHA triggering strands and many bivalent DNAzymes. The signal hairpins on the electrode are then cyclically cleaved by DNAzyme amplification cascade to liberate plenty G-quadruplex sequences, which bind hemin and yield largely enhanced currents for sensitive assay of MT with a detection limit of 0.217 nM in a label-free approach. Such sensor also shows selective discrimination capability to MT against other interfering proteins and assay of MT in normal serums with dilution has also been verified, indicating its potential for highly sensitive detection of different heavy metal ion binding molecules for various application scenarios.

Principles of miRNA/miRNA* function in plant MIRNA processing.

MicroRNAs (miRNAs) are essential regulators of gene expression, defined by their unique biogenesis, which requires the precise excision of the small RNA from an imperfect fold-back precursor. Unlike their animal counterparts, plant miRNA precursors exhibit variations in sizes and shapes. Plant MIRNAs can undergo processing in a base-to-loop or loop-to-base direction, with DICER-LIKE1 (DCL1) releasing the miRNA after two cuts (two-step MIRNAs) or more (sequential MIRNAs). In this study, we demonstrate the critical role of the miRNA/miRNA* duplex region in the processing of miRNA precursors. We observed that endogenous MIRNAs frequently experience suboptimal processing in vivo due to mismatches in the miRNA/miRNA* duplex, a key region that fine-tunes miRNA levels. Enhancing the interaction energy of the miRNA/miRNA* duplex in two-step MIRNAs results in a substantial increase in miRNA levels. Conversely, sequential MIRNAs display distinct and specific requirements for the miRNA/miRNA* duplexes along their foldback structure. Our work establishes a connection between the miRNA/miRNA* structure and precursor processing mechanisms. Furthermore, we reveal a link between the biological function of miRNAs and the processing mechanism of their precursors with the evolution of plant miRNA/miRNA* duplex structures.

DinoKnot: Duplex Interaction of Nucleic Acids With PseudoKnots.

Interaction of nucleic acid molecules is essential for their functional roles in the cell and their applications in biotechnology. While simple duplex interactions have been studied before, the problem of efficiently predicting the minimum free energy structure of more complex interactions with possibly pseudoknotted structures remains a challenge. In this work, we introduce a novel and efficient algorithm for prediction of Duplex Interaction of Nucleic acids with pseudoKnots, DinoKnot follows the hierarchical folding hypothesis to predict the secondary structure of two interacting nucleic acid strands (both homo- and hetero-dimers). DinoKnot utilizes the structure of molecules before interaction as a guide to find their duplex structure allowing for possible base pair competitions. To showcase DinoKnots's capabilities we evaluated its predicted structures against (1) experimental results for SARS-CoV-2 genome and nine primer-probe sets, (2) a clinically verified example of a mutation affecting detection, and (3) a known nucleic acid interaction involving a pseudoknot. In addition, we compared our results against our closest competition, RNAcofold, further highlighting DinoKnot's strengths. We believe DinoKnot can be utilized for various applications including screening new variants for potential detection issues and supporting existing applications involving DNA/RNA interactions, adding structural considerations to the interaction to elicit functional information.

In-situ EBSD investigation of excellent strength-ductility synergy mechanism achieved by thermal-mechanical processing of Al0·25FeCoNiV high-entropy alloy

The development of high-entropy alloys (HEAs) is plagued by a trade-off between strength and ductility. In this study, we investigated the effects of different cold rolling ratios and subsequent heat treatments on the microstructures and mechanical properties of the Al0·25FeCoNiV HEA. An efficient and economical thermal-mechanical processing route was successfully explored to prepare multiscale, multiphase heterogeneous structures consisting of heterogeneous grain structures and duplex structures. Yield strength, ultimate tensile strength, and fracture elongation of 1277.3 MPa, 1633.6 MPa, and 26.4 %, respectively, could be achieved. The strength-ductility trade-off was avoided. Detailed information on the evolution of grain boundary distribution, grain orientation changes, and grain rotation under stress and strain fields were obtained with in-situ electron backscatter diffraction (EBSD) tensile tests. The mechanism of coordinated plastic deformation and the intrinsic mechanism of excellent strength and ductility were investigated by quantitatively calculating the activation of the slip system, slip transfer at the phase boundaries, and grain rotation paths. It was found that the excellent mechanical properties can be attributed to multistage hetero-deformation-induced (HDI) strengthening and strain-hardening coupling mechanisms. In this study, an effective method for designing novel HEAs with excellent strength-ductility combinations and an analytical method based on in-situ tests are proposed.

Compression and fretting wear studies of γ/α 2 duplex TiAl alloys at the nanoscale

The study investigated the nanofabrication behavior of TiAl alloys with a duplex structure of γ/α 2. The process included downward pressure followed by reciprocating friction with diamond grinding balls and was simulated using molecular dynamics (MD). It was found that a certain number of dislocations in the workpiece was low, and the resilience was high during the initial pressing stage. The dislocations increased, the resilience decreased, and the plastic deformation capacity was enhanced under continuous pressing. The α 2 phase did not deform significantly during the compression process. The presence of the α 2 phase increases the overall hardness of the material, and elastic-plastic deformation occurs mainly where the γ phase is present; the endowment layer dislocations generated during the intrinsic stacking fault rebound via the phase boundary to form V-shaped dislocations. During the reciprocating friction of the workpiece, forward friction produces V-shaped dislocations, and reverse friction makes the dislocations disappear. This process results in the forward average friction force being more significant than the reverse average. γ/α 2 phase boundary has an impeding effect on the downward proliferation of defects, and the phase boundary makes the temperature transfer appear discontinuous. During friction, the specific number of vacancy atoms in the γ-phase increases, and the transition between FCC and HCP occurs.

Multi-material stainless steel fabrication using plasma wire arc additive manufacturing

Layered stainless steel samples were manufactured using Plasma Wire Arc Additive Manufacturing by depositing alternating layers of two dissimilar stainless steels onto an AISI 316L substrate. The selected materials were the M430 ferritic and M316L austenitic stainless steels. Comprehensive microstructural characterization in different areas, including interfaces between the two different materials, was achieved through optical microscopy and electron backscatter diffraction. Results revealed good weldability among the two-layered steels and on the substrate. Also, a gradient duplex structure was observed mainly at the layer interfaces, suggesting a nuanced blending of properties in the deposited zone. To establish a direct correlation with mechanical properties in the observed structures, macro and micro-hardness tests were conducted along the cross-section. These tests evidenced the presence of a harder phase along the interface compared to the individual materials. The microstructural analysis corroborated the presence of mixing between phases in conjunction with some areas of delta ferrite.

Enhanced Sequence-Specific DNA Recognition Using Oligodeoxynucleotide-Benzimidazole Conjugates.

Synthetic modification of oligodeoxynucleotides (ODNs) via conjugation to nucleic acid binding small molecules can improve hybridization and pharmacokinetic properties. In the present study, five Hoechst 33258 derived benzimidazoles were conjugated to T rich ODNs and their hybridization effectiveness was tested. Thermal denaturation studies revealed significant stabilization of complementary duplexes by ODN-benzimidazole conjugates, with the extent of stabilization being highly dependent on the length of the linker between DNA and benzimidazole. The increases in thermal stability were determined to be due to the binding of the benzimidazole moiety to the duplex. Circular dichroism and molecular modeling studies provided insights toward the influence of conjugation on duplex structure and how linker length impacts placement of the benzimidazole moiety in the minor groove. Furthermore, thermal denaturation studies with the complementary strand containing a single base mismatch or being RNA revealed that covalent conjugation of benzimidazoles to an ODN also enhances the sequence specificity. The fundamental studies reported herein provide a strategy to improve the stability and specificity properties of the ODN probes, which can be of use for targeting and diagnostics applications.