Strand Formation Research Articles

Physical Properties of Claudin-15 Strands in Tight Junctions

Tight junction claudins seal the paracellular pathway between two adjacent cells by forming selective permeation pathways. These barriers comprise a network of sealing strands in the cell membrane of one cell associating laterally with strands in the membrane of an adjacent cell. Tight junction strands are dynamic and maintain their barrier function during cell movements and large-scale tissue rearrangements. Strand formation requires coordinated claudin side-by-side (cis-) interactions within the same membrane and head-to-head (trans-) interactions between monomers from adjoining cells. To study the flexibility of claudin strands and the protein-protein interactions that provide this flexibility, we have constructed large-scale hybrid models of claudin-15 strands of various lengths ranging from 35nm to 215nm in their native environments. In this model, using the PACE forcefield, the protein is represented with atomic resolution and lipids and solvent molecules are coarse-grained using the MARTINI forcefield to reduce the computational cost. We have used molecular dynamics simulation to characterize physical properties of claudin strands including their bending modulus and persistence length from equilibrium simulations. In addition, we have identified the side-by-side (cis-) interactions in claudin-15 that are responsible for maintaining the strand conformation and providing flexibility. Our studies provide novel molecular insight into the mechanisms of claudin strand formation which is key to understanding how claudins interact and maintain selectively permeable paracellular barriers.

Cutting antiparallel DNA strands in a single active site.

SummaryA single enzyme active site that catalyzes multiple reactions is a well-established biochemical theme, but how one nuclease site cleaves both DNA strands of a double helix has not been well understood. In analyzing site-specific DNA cleavage by the mammalian RAG1-RAG2 recombinase, which initiates V(D)J recombination, we find that the active site is reconfigured for the two consecutive reactions, and the DNA double helix adopts drastically different structures. For initial nicking of the DNA, a locally unwound and unpaired DNA duplex forms a zipper via alternating inter-strand base stacking, rather than melting as generally thought. The second strand cleavage and formation of a hairpin-DNA product requires a global scissor-like movement of protein and DNA, delivering the scissile phosphate into the rearranged active site.

Morphological and histological analysis of anther-derived embryos of coconut (Cocos nucifera L.)

Doubled-haploids are a great source of material for heterozygous coconuts to shorten the time taken for crop improvement through hybridization programs. However, low frequency of embryo conversion and formation of weak plantlets are the major limitations in the protocol. In the present study anther-derived embryos were analysed at cellular level through the histological observations to understand the occurrence of shoot differentiation in those embryos. Attempts were also made to optimise the 6-Benzylaminopurine (BAP) concentration in the regeneration medium. Among the tested BAP levels (5, 15, 25, 35, 45, 55 μM), the regeneration medium containing 25 and 35 μM gave rise to 51.5% sprouted embryos against the control that gave only 22.7%. Among the analysed anther-derived embryos sequential events of differentiation including the formation of provascular strands followed by vascular bundle, differentiation in to the growing point, giving rise to the secondary embryos and polarization in to the shoot and root pole were identified. The blunt embryos devoid any morphological sign of sprouting showed different cellular arrangement viz. haustorial structure without any meristematic point (47%), bipolar with shoot and root meristems (8%) and unipolar with either of the pole (45%). Thick haustorial cover lead to the physical dormancy inhibiting further development of the polar structures. The germinating embryos containing a single shoot gave rise to the healthy plants where as double, multiple or fused shoots formed the week plantlets. The findings can be used for further optimisation of the protocol to achieve a greater plant regeneration frequency. 1. Culture media supplemented with increased levels of 6-Benzylaminopurine (BAP) enhanced the regeneration frequency of anther-derived embryos. 2. Thick haustorial tissue created an obstacle effect on further development of the differentiated shoot apical meristem. 3. Formation of clustered plants through secondary embryogenesis or shoot multiplication at the early stage of plant regeneration caused to give rise to the week plants.

3D bioprinting of anisotropic engineered tissue constructs with ultrasonically induced cell patterning

As 3D bioprinting continues to evolve as a promising alternative to engineer complex human tissues in-vitro, there is a need to augment bioprinting processes to achieve the requisite cellular and extracellular organizational characteristics found in the original tissues. While the cell distribution within bioinks is typically homogeneous, incorporating appropriate cellular patterning within the bioprinted constructs is an essential first step towards the eventual formation of anisotropically organized tissue matrix essential to its biomechanical form and function. This study describes a new bioprinting technique that uses ultrasonic standing bulk acoustic waves (SBAW) to organize cells into controllable anisotropic patterns within viscous bioinks while maintaining high cell viability. First, we develop a 3D computational model to discern the SBAW pressure pattern in response to multiple ultrasonic frequencies (0.71–2 MHz). We then experimentally analyze the patterns and viabilities of human adipose-derived stem cells (hASC) and human osteosarcoma cells (MG63) in alginate as a function of the SBAW frequency. Computational results indicate the formation of parallel pressure strands with higher pressure amplitudes near the bottom of the deposited layer, which is corroborated by experimental images of cell patterning. The inter-strand spacing is found to be affected by the frequency (p < 0.0001), while an interaction effect between the cell type and frequency governs the width of the strands (p = 0.02). Further, we demonstrate the synergistic bioprinting and SBAW-induced patterning of hASC within alginate and gelatin methacrylate (GelMA) constructs in tandem with chemical and photo-crosslinking, respectively. Pertinent cellular patterning and viability of at least 80 % were noted in the alginate and GelMA constructs across the experimental design space. Finally, we demonstrate the vat photo-polymerization-based bioprinting of a 3-layered GelMA construct with hASC strand lay pattern of 0-45-90° across the layers. This work represents a step forward in advancing bioprinting capabilities to achieve biomimetic tissue constructs.

An enzyme-free electrochemical sandwich DNA assay based on the use of hybridization chain reaction and gold nanoparticles: application to the determination of the DNA of Helicobacter pylori.

An ultrasensitive enzyme-free electrochemical sandwich DNA biosensor is described for the detection of ssDNA oligonucleotides. A DNA sequence derived from the genom of Helicobacter pylori was selected as a model target DNA. The DNA assay was realized through catching target DNA on capture DNA immobilized gold electrode; then labeling the target DNA with reporter DNA (rpDNA) and initiator DNA (iDNA) co-modified gold nanoparticles (AuNPs). The high density of iDNAs serves as one of the amplification strategies. The iDNA triggers hybridization chain reaction (HCR) between two hairpins. This leads to the formation of a long dsDNA concatamer strand and represents one amplification strategy. The electrochemical probe [Ru(NH3)5L]2+, where L stands for 3-(2-phenanthren-9-ylvinyl)pyridine, intercalated into dsDNA chain. Multiple probe molecules intercalate into one dsDNA chain, serving as one amplification strategy. The electrode was subjected to differential pulse voltammetry for signal acquisition, and the oxidation peak current at -0.28V was recorded. On each AuNP, 240 iDNA and 25 rpDNA molecules were immobilized. Successful execution of HCR at the DNA-modified AuNPs was confirmed by gel electrophoresis and hydrodynamic diameter measurements. Introduction of HCR significantly enhances the DNA detection signal intensity. The assay has two linear ranges of different slopes, one from 0.01 fM to 0.5 fM; and one from 1 fM to 100 fM. The detection limit is as low as 0.68 aM. Single mismatch DNA can be differentiated from the fully complementary DNA. Conceivably, this highly sensitive and selective assay provides a general method for detection of various kinds of DNA. Graphical abstractSchematic representation of the detection and the amplification principles of the electrochemical sandwich DNA assay. Purple curl: Captured DNA; Green curl: Reporter DNA; Orange curl: HCR initiator DNA; Yellow solid-circle: Gold nanoparticle; H1 and H2: Two hairpin DNA; [Ru(NH3)5L]2+: Signal probe.

Regioselective formation of RNA strands in the absence of magnesium ions.

The oligomerization of ribonucleotides can produce short RNA strands in the absence of enzymes. This reaction gives one of two regioisomeric phosphodiester linkages, a 2′,5′- or a 3′,5′-diester. The former, non-natural linkage is detrimental for duplex stability, and is known to form preferentially in oligomerizations occurring in homogeneous solution with preactivated nucleotides in the presence of magnesium cations. We have studied ribonucleotide oligomerization with in situ activation, using NMR as monitoring technique. Unexpectedly, the known preference for 2′,5′-linkages in the oligomerization of AMP was reversed in the absence of magnesium ions at slightly basic pH. Further, oligomerization was surprisingly efficient in the absence of Mg2+ salts, producing oligomers long enough for duplex formation. A quantitative systems chemistry analysis then revealed that the absence of magnesium ions favors the activation of nucleotides, and that the high concentration of active species can compensate for slower coupling. Further, organocatalytic intermediates can help to overcome the unfavorable regioselectivity of the magnesium-catalyzed reactions. Our findings allay concerns that RNA may have been difficult to form in the absence of enzymes. They also show that there is an efficient path to genetic material that does not require mineral surfaces or cations known to catalyze RNA hydrolysis.

Linear Prediction of Nucleotides in a Genome Sequence

Nucleotides are organic molecules, which are monomer units that form polymers of nucleic acid ‘deoxyribonucleic acid (DNA)’ and ‘ribonucleic acid (RNA)’. The four nucleotides A, T, G and C get connected by phosphodiester bonds to form strands. Strand formation depends on innumerable factors related to inter and intra cellular parameters and functions. One cannot precisely say that a particular strand gets formed using such and such rules. The infinite possibilities of strand formation cannot be determined experimentally or in the framework of classical genetics. One can alternatively formulate a notion of the “Language of Genomes” wherein one can finitely specify infinite strands. This paper introduces a novel prediction algorithm, which generates possible strands based on available nucleotides statistics.

An ultrasensitive guanine wire-based resonance light scattering method using G-quadruplex self-assembly for determination of microRNA-122.

An enzyme-free resonance light scattering (RLS) method is described for the determination of microRNA-122. A guanine nanowire (G-wire) is used that consists of a predesigned DNA1 and a G-quadruplex sequence DNA2. These hybridize with microRNA-122 and partially hybridize with DNA2. After formation of stable double strands with DNA1, DNA2 is released. On addition of K+ and Mg2+ ions, the G-quadruplex sequences undergo self-assembly to form long filamentous G-wires. This increases the intensity of RLS. A 6.1 pM detection limit was obtained, and the linear response covers the 50 pM to 300nM microRNA concentration range. The method was successfully applied to the quantitation of microRNA-122 in hepatocellular carcinoma cell lysates. Conceivably, this assay can be extended to other RLS methods for biomarker detection by simply changing the sequence of DNA1. Graphical abstract The G-quadruplex sequences of DNA2 were locked with DNA1. The G-quadruplex fragments of DNA2 were released after the hybridization of microRNA-122 with DNA1. These liberated G-quadruplex sequences were self-assembled into long filamentous guanine nanowires (G-wires) which increased resonance light intensity in the presence of Mg2+.

Effects of Arginine Deimination and Citrulline Side-Chain Length on Peptide Secondary Structure Formation.

Post-translational modifications expand the chemical functionality of peptides and proteins beyond that originating from the encoded amino acids, but studies on the structural effects of these modifications have been limited. Arginine undergoes deimination to give citrulline (Cit), converting the positively charged guanidinium moiety into a neutral urea group. Herein, we report the effect of Arg deimination on secondary structure formation. To understand the reason for the number of methylene units in Cit, the effect of Cit side-chain length on secondary structure formation was also studied. Ala-based peptides and β-hairpin peptides were used to study α-helix and β-sheet formation, respectively. Peptides containing Cit analogues were prepared by an orthogonal protecting group strategy coupled with solid-phase carbamylation. The CD data for the Ala-based peptides were analyzed by using modified Lifson-Roig theory, showing that the helix propensity of Arg decreased upon deimination and that either shortening or lengthening Cit also decreased the helix propensity. The β-hairpin peptides were analyzed by NMR methods, showing minimal change in strand formation energetics upon Arg deimination. Altering the Cit side-chain length did not affect strand formation energetics either. These results should be useful for the preparation of urea-bearing systems and the design of peptides incorporating urea-bearing residues with varying side-chain length.

1-nm-Wide Hydrated Dipole Arrays Regulate AuNW Assembly on Striped Monolayers in Nonpolar Solvent

Summary We show that striped phases of horizontally oriented phospholipids presenting 1-nm-wide orientable dipole arrays can order and straighten flexible 2-nm-diameter gold nanowires (AuNWs) with lengths (up to 1 μm) that greatly exceed the template pitch (∼7.5 nm). AuNW ordering can extend over areas > 100 μm2. Whereas wires bundle in solution, with contact between ligand shells, wires adsorbing to the template separate to much larger center-to-center distances (e.g., 12–30 nm) near multiples of the template pitch, consistent with repulsive interactions between wires emerging during interactions with dipole arrays in the template. AuNW assembly was carried out in cyclohexane, a nonpolar solvent. Interestingly, we found that wire ordering was contingent on the extent of phospholipid headgroup hydration and the presence of excess oleylamine, which assembled into hemicylindrical micelles around the hydrated headgroups. Together, these components generated a protected polar environment that enabled the phospholipid headgroups to function collectively to regulate AuNW assembly.

A Cellular Insulator against CLE45 Peptide Signaling

Plants continuously elaborate their bodies through post-embryonic, reiterative organ formation by apical meristems [1]. Meristems harbor stem cells, which produce daughter cells that divide repeatedly before they differentiate. How transitions between stemness, proliferation, and differentiation are precisely coordinated is not well understood, but it is known that phytohormones as well as peptide signals play important roles [2-7]. For example, in Arabidopsis thaliana root meristems, developing protophloem sieve elements (PPSEs) express thesecreted CLAVATA3/EMBRYO SURROUNDING REGION-RELATED 45 (CLE45) peptide and its cognate receptor, the leucine-rich repeat receptor kinase (LRR-RK) BARELY ANY MERISTEM 3 (BAM3). Exogenous CLE45 application or transgenically increased CLE45 dosage impairs protophloem formation, suggesting autocrine inhibition of PPSE differentiation by CLE45 signaling. Since CLE45 and BAM3 are expressed throughout PPSE development, it remains unclear how this inhibition is eventually overcome. The OCTOPUS (OPS) gene is required for proper PPSE differentiation and therefore the formation of continuous protophloem strands. OPS dosage increase can mend the phenotype of other mutants that display protophloem development defects in association with CLE45-BAM3 hyperactivity [8, 9]. Here, we provide evidence that OPS protein promotes differentiation of developing PPSEs by dampening CLE45 perception. This markedly quantitative antagonism is likely mediated through direct physical interference of OPS with CLE45 signaling component interactions. Moreover, hyperactive OPS confers resistance to other CLE peptides, and ectopic OPS overexpression triggers premature differentiation throughout the root. Our results thus reveal a novel mechanism in PPSE transition toward differentiation, wherein OPS acts as an "insulator" to antagonize CLE45 signaling.

Identification of a missense variant in CLDN2 in obstructive azoospermia.

Obstructive azoospermia (OA), defined as an obstruction in any region of the male genital tract, accounts for 40% of all azoospermia cases. Of all OA cases, ~30% are thought to have a genetic origin, however, hitherto, the underlying genetic etiology of the majority of these cases remain unknown. To address this, we took a family-based whole-exome sequencing approach to identify causal variants of OA in a multiplex family with epidydimal obstruction. A novel gain-of-function missense variant in CLDN2 (c.481G>C; p.Gly161Arg) was found to co-segregate with the phenotype, consistent with the X-linked inheritance pattern observed in the pedigree. To assess the pathogenicity of this variant, the wild and mutant protein structures were modeled and their potential for strand formation in multimeric form was assessed and compared. The results showed that dimeric and tetrameric arrangements of Claudin-2 were not only reduced, but were also significantly altered by this single residue change. We, therefore, envisage that this amino acid change likely forms a polymeric discontinuous strand, which may lead to the disruption of tight junctions among epithelial cells. This missense variant is thus likely to be responsible for the disruption of the blood-epididymis barrier, causing dislodged epithelial cells to clog the genital tract, hence causing OA. This study not only sheds light on the underlying pathobiology of OA, but also provides a basis for more efficient diagnosis in the clinical setting.

Anticancer Activity and Topoisomerase II Inhibition of Naphthalimides with ω-Hydroxylalkylamine Side-Chains of Different Lengths.

The substituted 1,8-Naphthalimides (1H-benzo[de]isoquinoline-1,3(2H)- diones) are known as DNA intercalators stabilizing DNA-Topoisomerase II complexes. This interaction disrupts the cleavage-relegation equilibrium of Topo II, resulting in formation of broken strands of DNA. To investigate the influence of type of substituents and substitution positions in 1,8- naphthalimde skeleton on the inhibition of Topoisomerase II activity. The starting 1,8-naphthalimide were prepared from acenaphthene by introduction of appropriate substituents followed by condensation with ω-hydroxylakylamines of different chain length. The substituents were introduced to 1,8-naphthalimide molecule by nucleophilic substitution of leaving groups like nitro or bromo present in 4 or 4,5- positions using the ω- hydroxylalkylamines. The bioactivity of obtained compounds was examined in model cell lines. Antiproliferative activity of selected compounds against HCT 116 human colon cancer cells, human non-small cell lung cells A549 and non-tumorigenic BEAS-2B human bronchial epithelium cells was examined. Several of investigated compounds exhibit a significant activity (IC50 µM to 7 µM) against model cancer cell lines. It was demonstrated that upon treatment with concentration of 200 µM, all derivatives display Topo II inhibitory activity, which may be compared with activity of Amonafide. The replacement of the nitro groups in the chromophore slightly reduces its anticancer activities, whereas the presence of both nitro group and ω-hydroxylalkylamine chain resulted in seriously increased anticancer activity. Obtained compounds showed Topo II inhibitory activity, moreover, influence of the substitution pattern on the ability to inhibit Topo II activity and cancer cells proliferation was observed.

Recovery Processes in the Adrenal Glands of Dogs

The aim of this research was a comparative study of cyclic changes in secretory, destructive and proliferative processes in the adrenal cortex in dogs. Material and methods . Cyclic changes of morphofunctional indices of adrenal cortex in dogs (Canis familiaris) were studied. Healthy mature males were used. The material was collected in the period from December to April with a monthly interval. For light microscopy the fragments of the adrenal glands were fixed in Steve, Bouin and Zenker liquids, 10% neutral formalin solution. PAS-reaction was used. RNA was detected by the Brachet reaction and gallocyanine according to Einarsson. The material fixed in a 10% solution of neutral formalin was used to study lipids. Results. In January, in the adrenal cortex clearly expressed glomerular, fasciculate and reticulae zones. In the glomerular zone elongated cells with oval nuclei are situated, in the fascicular - polygonal cells with rounded nuclei. In February, a decrease in the functional activity of the cortical parenchyma and the development of extensive destructive changes in glandular cells, especially in the glomerular zone, were found in the adrenal cortex. In March, the development of recovery processes and increased secretory activity of the adrenal cortex were noted. Proliferative processes are accompanied by loosening of the capsule, the formation of connective tissue strands going into the fascicular zone. Among the fibers of connective tissue fibroblasts, fibrocytes and small poorly differentiated cells with rounded nuclei are disposed. These cells are converted into elongated cells of the glomerular zone and polygonal cells of the fascicular zone.

Resolving the structure of organic nano strands self-assembled at a graphite-liquid interface using STM

Scanning tunneling microscopy (STM) at solution-graphite(0001) interface is used to identify nanometer-wide elementary strands of custom-designed ampiphilic benzamides and elucidate their internal structure with sub-molecular resolution. Evidences against graphitic artifacts that often mimic organic strands are provided, thereby unambiguously establishing the molecular origin of these strands. The aliphatic chain lengths are chosen based on bulk studies so as to promote strand architectures and avoid monolayer structures. Two different chain lengths are used to decipher the structural parameters and the results suggest hitherto unknown precursor routes to strand formation on a surface that is different from columnar mesophases in bulk. An on-surface self-assembly into hydrogen-bonded tetramer precursors and their subsequent interaction with other units via van der Waals forces between the dangling alkyl chains is proposed for strand formation on the surface.

Morphologic determinant of tight junctions revealed by claudin-3 structures

Tight junction is a cell adhesion apparatus functioning as barrier and/or channel in the paracellular spaces of epithelia. Claudin is the major component of tight junction and polymerizes to form tight junction strands with various morphologies that may correlate with their functions. Here we present the crystal structure of mammalian claudin-3 at 3.6 Å resolution. The third transmembrane helix of claudin-3 is clearly bent compared with that of other subtypes. Structural analysis of additional two mutants with a single mutation representing other subtypes in the third helix indicates that this helix takes a bent or straight structure depending on the residue. The presence or absence of the helix bending changes the positions of residues related to claudin-claudin interactions and affects the morphology and adhesiveness of the tight junction strands. These results evoke a model for tight junction strand formation with different morphologies – straight or curvy strands – observed in native epithelia.

Non-Cell-Autonomous Function of the GPI-Anchored Protein Undicht during Septate Junction Assembly

Occluding cell-cell junctions are pivotal during the development of many organs. One example is septate junction (SJ) strands, which are found in vertebrates and invertebrates. Although several proteins have been identified that are responsible for septate junction formation in Drosophila, it is presently unclear how these structures are formed or how they are positioned in a coordinated manner between two neighboring cells and within the tissue. Here, we identified a GPI-anchored protein called Undicht required for septate junction formation. Clonal analysis and rescue experiments show that Undicht acts in a non-cell-autonomous manner. It can be released from the plasma membrane by the proteolytic activity of two related ADAM10-like proteases, Kuzbanian and Kuzbanian-like. We propose that juxtacrine function of Undicht coordinates the formation of septate junction strands on two directly neighboring cells, whereas paracrine activity of Undicht controls the formation of occluding junctions within a tissue.

РОЛЬ ЦИТОСКЕЛЕТА ПРИ ДЕФОРМАЦИИ ЭРИТРОЦИТОВ В СТРЕССОВЫХ СИТУАЦИЯХ

The article presents the results of the action of cytochalazine B on the change in the topography of the membranes of red blood cells and the rearrangement of cytoskeleton elements using scanning electron microscopy. In the course of the studies, the degree of violation of the structure of membrane proteins, changes in the shape of cells and cytoskeleton elements, which are observed during castration of boars, were studied. The dynamics of changes in the architectonics of erythrocyte membranes is one of the characteristics of the etiopathogenesis of a number of disorders in the normal functioning of the individual. These changes were evaluated on blood samples using scanning electron microscopy. Red blood cells were pre-fixed under conditions excluding deformation of the cell surface. Dehydration in alcohol solutions of increasing concentration was carried out, dried by the transition of the critical point of CO2 in the HCP-Hitachi installation. During the study, changes in the surface relief of cell membranes characteristic of this cytostatic were observed. Under the action of the drug cytochalasin B, the formation of rare long strands with high optical density was observed when stained according to Heidenhain. The significant heterogeneity in the color intensity was noted, areas of dense hematoxylin staining were identified as well. The localization zones of actin are shown in the near-membrane regions after 30-45 minutes of cytochalazine action. The use of the drug Cytochalazine B in an isosmotic environment leads to an internal rearrangement of erythrocyte membrane proteins, and also determines a change in the shape of cells. This effect can be considered an analogue of the stress factor. Under simulated conditions, the mechanisms of membrane-destructive action on erythrocyte cytokarkas proteins are triggered, leading to a change in the surface of the membrane and, as a result, a violation of some parameters of the microvasculature. It can be assumed that this model of stress exposure can be used to assess the effect of stress factors on the phospholipid matrix of the blood erythrocyte membrane.

Colorimetric adenosine aptasensor based on DNA cycling amplification and salt-induced aggregation of gold nanoparticles

An aptamer based assay is described for the colorimetric detection of adenosine. The presence of adenosine triggers the deformation of hairpin DNA oligonucleotide (HP1) containing adenosine aptamer and then hybridizes another unlabeled hairpin DNA oligonucleotide (HP2). This leads to the formation of a double strand with a blunt 3' terminal. After exonuclease III (Exo III)-assisted degradation, the guanine-rich strand (GRS) is released from HP2. Hence, the adenosine-HP1 complex is released to the solution where it can hybridize another HP2 and initiate many cycles of the digestion reaction with the assistance of Exo III. This leads to the generation of a large number of GRS strands after multiple cycles. The GRS stabilize the red AuNPs against aggregation in the presence of potassium ions. If, however, GRS forms a G-quadruplex, it loses its ability to protect gold nanoparticles (AuNPs) from salt-induced AuNP aggregation. Therefore, the color of the solution changes from red to blue which can be visually observed. This colorimetric assay has a 0.13nM detection limit and a wide linear range that extends from 5nM to 1μM. Graphical abstract Schematic presentation of acolorimetric aptamer biosensor for adenosine detection based on DNA cycling amplification and salt-induced aggregation of gold nanoparticles.

Modeling of the Twin-Roll Casting Process: Transition from Casting to Rolling

During twin-roll casting, an alloy melt is passing the gap between two counter-rotating rolls, where cooling and solidification leads to the continuous formation of a solid strand. In order to describe this process, a two-phase Eulerian–Eulerian volume-averaging model is presented that accounts for (1) transport and growth of spherical grains within a flowing melt, (2) the formation of a coherent solid network above a specific solid fraction and (3) the viscoplastic flow of the solid network with the interstitial melt during casting and compression. For the considered case of an inoculated Al–4wt%Cu alloy, the process conditions are chosen such that two relatively thick viscoplastic semi-solid shells meet between the rolls, and thus, the material is pressed together and squeezed against the casting direction. The squeezed out material consists of segregated melt and some solid that quickly disappears after melting. It is observed during this study that macrosegregation distributions are inherently connected to the mush deformation that is enforced during the hot rolling process.