Synergistic approaches to ultra-precision high performance cutting

Diamond milling allows for the flexible production of optical and high precision parts, but suffers from poor setup and production speeds. This paper presents recent advances that aim towards achieving high performance (HPC) and high speed cutting (HSC) in ultra-precision machining. After a short introduction, the benefits of high speed cutting for both metals and brittle-hard materials are shown. Thereafter, novel mechatronic devices are presented that enable an automated balancing of the applied air bearing spindles and the application of multiple diamond tools on one tool holder and by thus, contribute to HPC. These developments are supplemented by a novel linear guiding system based on electromagnatic levitation that, along with a dedicated model-based control system, enables fast and precise movements of the machine tool. After presenting the recent developments in detail, their synergistic performance is assessed and an outlook to future developments is given. © 2020 The Author

Reconstitution of CPSF active in polyadenylation: recognition of the polyadenylation signal by WDR33

Cleavage and polyadenylation specificity factor (CPSF) is the central component of the 3' processing machinery for polyadenylated mRNAs in metazoans: CPSF recognizes the polyadenylation signal AAUAAA, providing sequence specificity in both pre-mRNA cleavage and polyadenylation, and catalyzes pre-mRNA cleavage. Here we show that of the seven polypeptides that have been proposed to constitute CPSF, only four (CPSF160, CPSF30, hFip1, and WDR33) are necessary and sufficient to reconstitute a CPSF subcomplex active in AAUAAA-dependent polyadenylation, whereas CPSF100, CPSF73, and symplekin are dispensable. WDR33 is required for binding of reconstituted CPSF to AAUAAA-containing RNA and can be specifically UV cross-linked to such RNAs, as can CPSF30. Transcriptome-wide identification of WDR33 targets by photoactivatable ribonucleoside-enhanced cross-linking and immunoprecipitation (PAR-CLIP) showed that WDR33 binds in and very close to the AAUAAA signal in vivo with high specificity. Thus, our data indicate that the large CPSF subunit participating in recognition of the polyadenylation signal is WDR33 and not CPSF160, as suggested by previous studies

Diamond Micro Chiseling of large-scale retroreflective arrays

Triple mirror retroreflectors are essential components for safety applications, communications and measurement equipment. While downscaling of characteristic dimension is possible for triangular retroreflectors, this is a challenging task for full-cube retroreflectors, due to the absence of continuous tool paths. Thus, the Diamond Micro Chiseling (DMC) process has been developed which allows the machining of full-cube retroreflectors by overlapping a series of sharp-edged pyramidal microcavities. In the past, this has been successfully demonstrated on a small-scale up to 3 mm × 3 mm with a structure size of 150 μm. Industrial applications, however, require the structuring of areas which are significantly larger than 10 mm × 10 mm. This paper will introduce the technology for machining such pattern with the help of the DMC process. Particular attention will be given to the measurement procedures and required tolerances for performing an in situ tool change as well as the optimization strategies for reducing the required process time.65065736

Sequential Processing of Cold Gas Sprayed Alloys by Milling and Deep Rolling

Cold gas spraying (CS) is a solid-state material deposition process which, in addition to the flexible repair of individual component areas, also enables the build-up of larger samples. The layers are created on a substrate by the impact-induced bonding of highly accelerated micrometer particles. Since melting does not occur, the material composition can be varied flexibly and independently of material-specific melting points. In this work, the influence of the described forming process on subsequent machining by milling and deep rolling is investigated. The process forces measured during milling and the surface topography after milling and deep rolling were influenced by the material composition and the CS-related properties, e.g., high material hardness or particle bonding. In contrast to prior assumptions, deep rolling was shown to have no influence on the determined hardness depth profile for the investigated materials. Future work will focus on additional analyses, such as the determination of half-widths, to obtain further insight on the material behavior

Crystal structure of a covalently linked Aurora-A-MYCN complex

Formation of the Aurora-A–MYCN complex increases levels of the oncogenic transcription factor MYCN in neuroblastoma cells by abrogating its degradation through the ubiquitin proteasome system. While some small-molecule inhibitors of Aurora-A were shown to destabilize MYCN, clinical trials have not been satisfactory to date. MYCN itself is considered to be `undruggable' due to its large intrinsically disordered regions. Targeting the Aurora-A–MYCN complex rather than Aurora-A or MYCN alone will open new possibilities for drug development and screening campaigns. To overcome the challenges that a ternary system composed of Aurora-A, MYCN and a small molecule entails, a covalently cross-linked construct of the Aurora-A–MYCN complex was designed, expressed and characterized, thus enabling screening and design campaigns to identify selective binders

Micro-Injection Molding of Diffractive Structured Surfaces

In recent years, the use of highly functional optical elements has made its way into our everyday life. Its applications range from use in utility items such as cell phone cameras up to security elements on banknotes or production goods. For this purpose, the Leibniz Institute for Materials Engineering (IWT) has been developing a cutting process for the fast and cost-effective production of hologram-based diffractive optical elements. In contrast to established non-mechanical manufacturing processes, such as laser lithography or chemical etching, which are able to produce optics in large quantities and with high accuracy, the diamond turning approach is extending these properties by offering several degrees of freedom. This allows for an almost unlimited geometric complexity and a structured area of considerable size (several tenth square millimeters), achieved in a single process step. In order to introduce diffractive security features to the mass market and to actual production goods, a high-performance replication process is required as the consecutive development step. Micro injection molding represents a feasible and promising option here. In particular, diamond machining enables the integration of safety features directly into the mold insert. Not only does this make additional assembly obsolete, but the safety feature can also be placed inconspicuously in the final product. In this paper, the potential of micro-injection molding as a replication process for diffractive structured surfaces will be investigated and demonstrated. Furthermore, the optical functionality after replication will be verified and evaluated

Micro-Injection Molding of Diffractive Structured Surfaces

In recent years, the use of highly functional optical elements has made its way into our everyday life. Its applications range from use in utility items such as cell phone cameras up to security elements on banknotes or production goods. For this purpose, the Leibniz Institute for Materials Engineering (IWT) has been developing a cutting process for the fast and cost-effective production of hologram-based diffractive optical elements. In contrast to established non-mechanical manufacturing processes, such as laser lithography or chemical etching, which are able to produce optics in large quantities and with high accuracy, the diamond turning approach is extending these properties by offering several degrees of freedom. This allows for an almost unlimited geometric complexity and a structured area of considerable size (several tenth square millimeters), achieved in a single process step. In order to introduce diffractive security features to the mass market and to actual production goods, a high-performance replication process is required as the consecutive development step. Micro injection molding represents a feasible and promising option here. In particular, diamond machining enables the integration of safety features directly into the mold insert. Not only does this make additional assembly obsolete, but the safety feature can also be placed inconspicuously in the final product. In this paper, the potential of micro-injection molding as a replication process for diffractive structured surfaces will be investigated and demonstrated. Furthermore, the optical functionality after replication will be verified and evaluated

Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors

Cleavage factor II (CF II) is a poorly characterized component of the multiprotein complex catalyzing 3' cleavage and polyadenylation of mammalian mRNA precursors. We have reconstituted CF II as a heterodimer of hPcf11 and hClp1. The heterodimer is active in partially reconstituted cleavage reactions, whereas hClp1 by itself is not. Pcf11 moderately stimulates the RNA 5' kinase activity of hClp1; the kinase activity is dispensable for RNA cleavage. CF II binds RNA with nanomolar affinity. Binding is mediated mostly by the two zinc fingers in the C-terminal region of hPcf11. RNA is bound without pronounced sequence-specificity, but extended G-rich sequences appear to be preferred. We discuss the possibility that CF II contributes to the recognition of cleavage/polyadenylation substrates through interaction with G-rich far-downstream sequence elements