Assembly Of Components Research Articles

Gephyrin promotes autonomous assembly and synaptic localization of GABAergic postsynaptic components without presynaptic GABA release

Synapses containing γ-aminobutyric acid (GABA) constitute the primary centers for inhibitory neurotransmission in our nervous system. It is unclear how these synaptic structures form and align their postsynaptic machineries with presynaptic terminals. Here, we monitored the cellular distribution of several GABAergic postsynaptic proteins in a purely glutamatergic neuronal culture derived from human stem cells, which virtually lacks any vesicular GABA release. We found that several GABAA receptor (GABAAR) subunits, postsynaptic scaffolds, and major cell-adhesion molecules can reliably coaggregate and colocalize at even GABA-deficient subsynaptic domains, but remain physically segregated from glutamatergic counterparts. Genetic deletions of both Gephyrin and a Gephyrin-associated guanosine di- or triphosphate (GDP/GTP) exchange factor Collybistin severely disrupted the coassembly of these postsynaptic compositions and their proper apposition with presynaptic inputs. Gephyrin-GABAAR clusters, developed in the absence of GABA transmission, could be subsequently activated and even potentiated by delayed supply of vesicular GABA. Thus, molecular organization of GABAergic postsynapses can initiate via a GABA-independent but Gephyrin-dependent intrinsic mechanism.

Redundant non-serial compound manipulator kinematics and dynamics

Redundant non-serial compound manipulators with closed kinematic and actuator loops that enhance precision control and load carrying capacity are treated. Generalized coordinates and associated holonomic constraints that account for kinematic and actuator closed loops and singularity free representation of orientation of spatial manipulator bodies augment input and output coordinates in the kinematic formulation. Disjoint assembly components and path connected singularity free assembly components of manipulator configuration space are defined, in which manipulator kinematics and dynamics can be reliably carried out. A set-valued inverse kinematic configuration mapping is derived, with which compound redundant manipulators are shown to be locally cyclic. A new system of manipulator operational coordinates comprised of output and self-motion coordinates are defined and shown to be equivalent to input coordinates in representation of manipulator kinematics and dynamics. Well-posed ordinary differential equations of manipulator dynamics in both input and operational coordinates are presented that require no ad-hoc derivation. Five model manipulators illustrate applications involved and theoretical methods presented.

Isolation and Partial Characterization of Novel, Structurally Uniform (Hfq6)n≥8 Assemblies Carrying Accessory Transcription and Translation Factors.

In growing E. coli cells, the transcription-translation complexes (TTCs) form characteristic foci; however, the exact molecular composition of these superstructures is not known with certainty. Herein, we report that, during our recently developed "fast" procedures for purification of E. coli RNA polymerase (RP), a fraction of the RP's α/RpoA subunits is displaced from the core RP complexes and copurifies with multiprotein superstructures carrying the nucleic acid-binding protein Hfq and the ribosomal protein S6. We show that the main components of these large multiprotein assemblies are fixed protein copy-number (Hfq6)n≥8 complexes; these complexes have a high level of structural uniformity and are distinctly unlike the previously described (Hfq6)n "head-to-tail" polymers. We describe purification of these novel, structurally uniform (Hfq6)n≥8 complexes to near homogeneity and show that they also contain small nonprotein molecules and accessory S6. We demonstrate that Hfq, S6, and RP have similar solubility profiles and present evidence pointing to a role of the Hfq C-termini in superstructure formation. Taken together, our data offer new insights into the composition of the macromolecular assemblies likely acting as scaffolds for transcription complexes and ribosomes during bacterial cells' active growth.

A Visual Measurement Method for Deep Holes in Composite Material Aerospace Components.

The visual measurement of deep holes in composite material workpieces constitutes a critical step in the robotic assembly of aerospace components. The positioning accuracy of assembly holes significantly impacts the assembly quality of components. However, the complex texture of the composite material surface and mutual interference between the imaging of the inlet and outlet edges of deep holes significantly challenge hole detection. A visual measurement method for deep holes in composite materials based on the radial penalty Laplacian operator is proposed to address the issues by suppressing visual noise and enhancing the features of hole edges. Coupled with a novel inflection-point-removal algorithm, this approach enables the accurate detection of holes with a diameter of 10 mm and a depth of 50 mm in composite material components, achieving a measurement precision of 0.03 mm.

Conserved cysteine residues in Kaposi's sarcoma herpesvirus ORF34 are necessary for viral production and viral pre-initiation complex formation.

Kaposi's sarcoma herpesvirus (KSHV) ORF34 plays a significant role as a component of the viral pre-initiation complex (vPIC), which is indispensable for late gene expression across beta and gamma herpesviruses. Although the key role of ORF34 within the vPIC and its function as a hub protein have been recognized, further clarification regarding its specific contribution to vPIC functionality and interactions with other components is required. This study employed a deep-learning algorithm-assisted structural model of ORF34, revealing highly conserved amino acid residues across human beta- and gamma-herpesviruses localized in structured domains. Thus, we engineered ORF34 alanine-scanning mutants by substituting conserved residues with alanine. These mutants were evaluated for their ability to interact with other vPIC factors and restore viral production in cells harboring the ORF34-deficient KSHV-BAC. Our experimental results highlight the crucial role of the 4 cysteine residues conserved in ORF34: a tetrahedral arrangement consisting of a pair of C-Xn-C consensus motifs. This suggests the potential incorporation of metal cations in interacting with ORF24 and ORF66 vPIC components, facilitating late gene transcription, and promoting overall virus production by capturing metal cations. In summary, our findings underline the essential role of conserved cysteines in KSHV ORF34 for effective vPIC assembly and viral replication, thereby enhancing our understanding of the complex interplay between the vPIC components.

Mixing water, sugar, and lipid: Conformations of isolated and micro-hydrated glycolipids in the gas phase.

Both sugars and lipids are important biomolecular building blocks with exceptional conformational flexibility and adaptability to their environment. Glycolipids bring together these two molecular components in the same assembly and combine the complexity of their conformational landscapes. In the present study, we have used selective double resonance vibrational spectroscopy, in combination with a computational approach, to explore the conformational preferences of two glycolipid models (3-0-acyl catechol and guaiacol α-D-glucopyranosides), either fully isolated in the gas phase or controlled interaction with a single water molecule. We could identify the preferred conformation and structures of the isolated and micro-hydrated species and evidence of the presence of a strong water pocket, which may influence the conformational flexibility of such systems, even in less controlled environments.

Spectrin condensates provide a nidus for assembling the periodic axonal structure.

Coordinated assembly of individual components into higher-order structures is a defining theme in biology, but underlying principles are not well-understood. In neurons, α/β spectrins, adducin, and actinfilaments assemble into a lattice wrapping underneath the axonal plasma membrane, but mechanistic events leading to this periodic axonal structure (PAS) are unclear. Visualizing PAS components in axons as they develop, we found focal patches in distal axons containing spectrins and adducin (but sparse actin filaments) with biophysical properties reminiscent of biomolecular condensation. Overexpressing spectrin-repeats - constituents of α/β-spectrins - in heterologous cells triggered condensate formation, and preventing association of βII-spectrin with actin-filaments/membranes also facilitated condensation. Finally, overexpressing condensate-triggering spectrin repeats in neurons before PAS establishment disrupted the lattice, presumably by competing with innate assembly, supporting a functional role for biomolecular condensation. We propose a condensation-assembly model where PAS components form focal phase-separated condensates that eventually unfurl into a stable lattice-structure by associating with subplasmalemmal actin. By providing local 'depots' of assembly parts, biomolecular condensation may play a wider role in the construction of intricate cytoskeletal structures.

Topological and lattice-based AM optimization for improving the structural efficiency of robotic arms

The robotic arm is one of the vital components of robot assembly. The purpose of the robotic arm is to transmit power and conduct the desired motion, i.e., translation or rotation. Robotic limbs are designed and constructed to execute certain tasks with a high degree of speed, accuracy, and efficiency. This research focuses on to enhancing the strength-to-weight ratio of robotic arm using certain techniques of additive manufacturing, i.e., topology optimization and lattice structure. Employing the finite element analysis, the impact of weight reduction optimization on structural parameters such as stress and deformation in the current design is assessed using ANSYS R18.1 for FE analysis and Creo parametric 7.0 design software for computer-aided design modeling. Observations reveal that the 0.5 and .4 scale lattice structure designs have deformation of 0.01453mm and 0.01453 mm respectively though the generic design has 0.01043 mm deformation. Notably, the 0.5 scale lattice of the robotic arm exhibits a 31.08% higher equivalent stress than the generic design with 29.3%. reduction in mass of the robotic arm. These findings highlight the efficacy of lattice structures for optimizing the robotic arm’s performance, contributing to advancements in power-efficient robot assembly processes.

An Ergonomic Risk Assessment System Based on 3D Human Pose Estimation and Collaborative Robot

Human pose estimation focuses on methods that allow us to assess ergonomic risk in the workplace and aims to prevent work-related musculoskeletal disorders (WMSDs). The recent increase in the use of Industry 4.0 technologies has allowed advances to be made in machine learning (ML) techniques for image processing to enable automated ergonomic risk assessment. In this context, this study aimed to develop a method of calculating joint angles from digital snapshots or videos using computer vision and ML techniques to achieve a more accurate evaluation of ergonomic risk. Starting with an ergonomic analysis, this study explored the use of a semi-supervised training method to detect the skeletons of workers and to estimate the positions and angles of their joints. A criticality index, based on RULA scores and fuzzy rules, is then calculated to evaluate possible corrective actions aimed at reducing WMSDs and improving production capacity using a collaborative robot that supports workers in carrying out critical operations. This method is tested in a real industrial case in which the manual assembly of electrical components is conducted, achieving a reduction in overall ergonomic stress of 13% and an increase in production capacity of 33% during a work shift. The proposed approach can overcome the limitations of recent developments based on computer vision or wearable sensors by performing an assessment with an objective and flexible approach to postural analysis development.

Enhancement of the DUNE FD1 X-ARAPUCA Photon Detection Efficiency and upgrade of the FD2 Photon Collector

The Photon Detection System (PDS) of the first two DUNE far detectors (FD1 and FD2) is composed of 6000 and 672 photon detection units respectively, named X-Arapuca, of different size and geometry. The PDS will complement and boost the DUNE LArTPC for the detection of non beam events: the prompt light detection will improve their tagging, and at low energies it will enable the trigger and the calorimetry of the supernova neutrinos. The X-Arapuca unit is an assembly of several components: its Photon Detection Efficiency (PDE) depends both on the design of the assembly and on the grade and the coupling of the individual components. The X-Arapuca PDE is the driver of the Photon Detection System sensitivity, that in turn determines the sensitivity of the DUNE physics reach for the detection of core-collapse supernova within the galaxy and for nucleon decay searches. In this work we present an update of the absolute PDE of the FD1 X-Arapuca baseline design, measured in laboratory: 160 units of this are deployed in the scale 1:20 FD1 prototype hosted in the NP04 cryostat at the CERN neutrino platform. Further we show how to change the baseline design of the FD1 X-Arapuca, allowing to double its PDE. Finally we review a few selected features of the photon collector of the sixteen FD2 X-Arapuca recently deployed for the FD2 scale 1:20 prototype at CERN in the NP02 cryostat, and of the last six units that integrate the latest advancements.

K29-linked free polyubiquitin chains affect ribosome biogenesis and direct ribosomal proteins to the intranuclear quality control compartment

Ribosome assembly requires precise coordination between the production and assembly of ribosomal components. Mutations in ribosomal proteins that inhibit the assembly process or ribosome function are often associated with ribosomopathies, some of which are linked to defects in proteostasis. In this study, we examine the interplay between several yeast proteostasis enzymes, including deubiquitylases (DUBs) Ubp2 and Ubp14, and E3 ligases Ufd4 and Hul5, and we explore their roles in the regulation of the cellular levels of K29-linked unanchored polyubiquitin (polyUb) chains. Accumulating K29-linked unanchored polyUb chains associate with maturing ribosomes to disrupt their assembly, activate the ribosome assembly stress response (RASTR), and lead to the sequestration of ribosomal proteins at the intranuclear quality control compartment (INQ). These findings reveal the physiological relevance of INQ and provide insights into mechanisms of cellular toxicity associated with ribosomopathies.

Desain dan Fabrikasi Printed Circuit Board (PCB) Smartband untuk Mengukur Tanda Vital Pada Tubuh Manusia

This research addresses the design and fabrication of a Printed Circuit Board (PCB) for a smartband used to measure vital signs on the human body, including heart rate and body temperature. The main objective of this research is to produce a reliable and accurate device in daily health monitoring. The research process includes system design, design validation, PCB fabrication, component assembly, and testing and calibration. Key components, including heart rate and temperature sensors, were selected and integrated using PCB design software. PCB fabrication was performed through photolithography, etching, drilling, and plating processes. The test results show that the designed PCB meets the expected technical specifications.

Transcriptional control of the Cryptosporidium life cycle.

The parasite Cryptosporidium is aleading agent of diarrhoeal disease in young children, and a cause and consequence of chronic malnutrition1,2. There are no vaccines and only limited treatment options3. The parasite infects enterocytes, in which it engages in asexual and sexual replication4, both of which are essential to continued infection and transmission. However, their molecular mechanisms remain largely unclear5. Here we use single-cell RNA sequencing to reveal the gene expression programme of the entire Cryptosporidium parvum life cycle in culture and in infected animals. Diverging from the prevailing model6, we find support for only three intracellular stages: asexual type-I meronts, male gamonts and female gametes. We reveal a highly organized program for the assembly of components at each stage. Dissecting the underlying regulatory network, we identify the transcription factor Myb-M as the earliest determinant of male fate, in an organism that lacks genetic sex determination. Conditional expression of this factor overrides the developmental program and induces widespread maleness, while conditional deletion ablates male development. Both have a profound impact on the infection. A large set of stage-specific genes now provides the opportunity to understand, engineer and disrupt parasite sex and life cycle progression to advance the development of vaccines and treatments.

A digital twin-driven human-machine interactive assembly method based on lightweight multi-target detection and assembly feature generation

ABSTRACT In the field of industrial assembly, human-machine interactive assembly methods are frequently used. Lack of virtual and physical mapping, a convoluted guiding system, and low effect precision in the interactive process, a digital twin-driven human-machine interactive assembly method system is proposed as a solution to the mentioned issues. The YOLOv7-tiny lightweight model is used to perform accurate detection of parts. By incorporating attention modules into the backbone network, the feature extraction capability of the model in complicated assembly environments is enhanced. The assembly method proposed is validated using the assembly process of the reducer as an instance. The OpenCV method is employed to produce geometric reference features for parts. The experimental results show that the proposed assembly method can provide visual guidance for the assembly process, improve the traditional list-type assembly component retrieval method, solve the drawbacks of the pre-set assembly guidance in the guidance system that may not be able to adapt to the changes of the assembly results in the actual operation, and can accurately instruct novices how to assemble, which is characterised by easy implementation, low cost and high accuracy, and is of great significance for improving the success rate and assembly efficiency of human-machine interactive assembly.

An industrial edge computing architecture for Local Digital Twin

Cyber–Physical Production Systems usher in a new era of smart manufacturing, with Digital Twin being a pivotal concept driving their adoption. Most literature concentrates on global Digital Twin deployment, typically enterprise servers or the cloud. However, manufacturing relies on real-time control and decision-making, demanding swift system response. To address this, we introduce a novel Local Digital Twin architecture, bringing processing to the edge to meet strict timing constraints. We also introduce a Five-dimension GDT/LDTs model aligned with RAMI4.0 for practical application and integration. The LDT was implemented on a multinational company’s Brazilian subsidiary assembly line, predicting optimal assembly components. Machine learning techniques were employed, improving the First Pass Yield by 1.3 to 2.5% in a test machine, affirming the feasibility of the LDT approach.

AiroTouch: enhancing telerobotic assembly through naturalistic haptic feedback of tool vibrations.

Teleoperation allows workers to safely control powerful construction machines; however, its primary reliance on visual feedback limits the operator's efficiency in situations with stiff contact or poor visibility, hindering its use for assembly of pre-fabricated building components. Reliable, economical, and easy-to-implement haptic feedback could fill this perception gap and facilitate the broader use of robots in construction and other application areas. Thus, we adapted widely available commercial audio equipment to create AiroTouch, a naturalistic haptic feedback system that measures the vibration experienced by each robot tool and enables the operator to feel a scaled version of this vibration in real time. Accurate haptic transmission was achieved by optimizing the positions of the system's off-the-shelf accelerometers and voice-coil actuators. A study was conducted to evaluate how adding this naturalistic type of vibrotactile feedback affects the operator during telerobotic assembly. Thirty participants used a bimanual dexterous teleoperation system (Intuitive da Vinci Si) to build a small rigid structure under three randomly ordered haptic feedback conditions: no vibrations, one-axis vibrations, and summed three-axis vibrations. The results show that users took advantage of both tested versions of the naturalistic haptic feedback after gaining some experience with the task, causing significantly lower vibrations and forces in the second trial. Subjective responses indicate that haptic feedback increased the realism of the interaction and reduced the perceived task duration, task difficulty, and fatigue. As hypothesized, higher haptic feedback gains were chosen by users with larger hands and for the smaller sensed vibrations in the one-axis condition. These results elucidate important details for effective implementation of naturalistic vibrotactile feedback and demonstrate that our accessible audio-based approach could enhance user performance and experience during telerobotic assembly in construction and other application domains.

Left and Right‐Handed Light Reflection and Emission in Ultrathin Cellulose Nanocrystals Films with Printed Helicity

AbstractNatural polymers, particularly plant‐derived nanocelluloses, self‐organize into hierarchical structures, enabling mechanical robustness, bright iridescence, emission, and polarized light reflection. These biophotonic properties are facilitated by the assembly of individual components during evaporation, such as cellulose nanocrystals (CNCs), which exhibit a left‐handed helical pitch in a chiral nematic state. This work demonstrates how optically active films with pre‐programmed opposite handedness (left or right) can be constructed via shear‐induced twisted printing with clockwise and counter‐clockwise shearing vectors. The resulting large‐area thin films are transparent yet exhibit pre‐determined mirror‐symmetrical optical activity, enabling the distinction of absorbed and emitted circularly polarized light. This processing method allows for sequential printing of thin and ultrathin films with twisted layered organization and on‐demand helicity. The complex light polarization behavior is due to step‐like changes in linear birefringence within each deposited layer and circular birefringence, different from that of conventional CNC films as revealed with Muller matrix analysis. Furthermore, intercalating an achiral organic dye into printed structures induces circularly polarized luminescence while preserving high transmittance and controlled handedness. These results suggest that twisted sequential printing can facilitate the construction of chiroptical metamaterials with tunable circular polarization, absorption, and emission for optical filters, encryption, photonic coatings, and chiral sensors.

The anticancer potential of the CLK kinases inhibitors 1C8 and GPS167 revealed by their impact on the epithelial-mesenchymal transition and the antiviral immune response.

The diheteroarylamide-based compound 1C8 and the aminothiazole carboxamide-related compound GPS167 inhibit the CLK kinases, and affect the proliferation of a broad range of cancer cell lines. A chemogenomic screen previously performed with GPS167 revealed that the depletion of components associated with mitotic spindle assembly altered sensitivity to GPS167. Here, a similar screen performed with 1C8 also established the impact of components involved in mitotic spindle assembly. Accordingly, transcriptome analyses of cells treated with 1C8 and GPS167 indicated that the expression and RNA splicing of transcripts encoding mitotic spindle assembly components were affected. The functional relevance of the microtubule connection was confirmed by showing that subtoxic concentrations of drugs affecting mitotic spindle assembly increased sensitivity to GPS167. 1C8 and GPS167 impacted the expression and splicing of transcripts in pathways relevant to tumor progression, including MYC targets and the epithelial mesenchymal transition (EMT). Finally, 1C8 and GPS167 altered the expression and alternative splicing of transcripts involved in the antiviral immune response. Consistent with this observation, depleting the double-stranded RNA sensor DHX33 suppressed GPS167-mediated cytotoxicity on HCT116 cells. Our study uncovered molecular mechanisms through which 1C8 and GPS167 affect cancer cell proliferation as well as processes critical for metastasis.

DS-PACK: Tool assembly for the end-to-end support of controlled access human data sharing

The EU General Data Protection Regulation (GDPR) requirements have prompted a shift from centralised controlled access genome-phenome archives to federated models for sharing sensitive human data. In a data-sharing federation, a central node facilitates data discovery; meanwhile, distributed nodes are responsible for handling data access requests, concluding agreements with data users and providing secure access to the data. Research institutions that want to become part of such federations often lack the resources to set up the required controlled access processes. The DS-PACK tool assembly is a reusable, open-source middleware solution that semi-automates controlled access processes end-to-end, from data submission to access. Data protection principles are engraved into all components of the DS-PACK assembly. DS-PACK centralises access control management and distributes access control enforcement with support for data access via cloud-based applications. DS-PACK is in production use at the ELIXIR Luxembourg data hosting platform, combined with an operational model including legal facilitation and data stewardship.

FABRICATION OF ORDERS BY SCARA ROBOT WITH RFID TECHNOLOGY

In today’s world of automation, recognizing and processing parcels in a particular area of zone is a crucial task. One such task is pick-and place operation, which is well- known for its challenges. The main theme of this operation is to sort products into different areas based on their representative fields. To address this issue, we have a SCARA robot that is known for its liner momentum motion of 4 degrees of freedom. This robot is well-suited for pick-and- place operations and can perform operations in two axes. Moreover, it is cost-effective and efficient. The SCARA robot is connected to the Arduino Mega, which is run by servo motors and controllers. The fabrication of an Arduino based Parcel diverter system involves the creation of a system that uses an Arduino Mega microcontroller to sort and divert parcels based on their destination. The system is designed to take input from Radio frequency identification device that detects the parcels and direct them to their appropriate destination using a series of mechanical diverters. The fabrication process includes the design and assembly of the mechanical components, the programming of the micro controller, and the integration of the sensors. The resulting system provides a reliable and efficient way to sort and direct parcels in a variety of industrial and logistical applications. Key Words: Conveyor, Arduino Mega, IR/Thermal sensors, stepper motor, micro step driver SCARA robot, RFID system, DC Vacuum pump and DC Motor.