Versatile System Research Articles

Tetracistronic minigenomes elucidate a functional promoter for Ghana virus and unveils Cedar virus replicase promiscuity for all henipaviruses.

Henipaviruses are recognized as significant global health threats due to their high mortality rates and lack of effective vaccines or therapeutics. Due to the requirement for high biocontainment facilities, the scope of research which may be conducted on henipaviruses is limited. To address this challenge, we developed innovative tetracistronic, transcription, and replication-competent minigenomes. We demonstrate that these systems replicate key aspects of the viral life cycle, such as budding, fusion, and receptor binding, and are safe for use in lower biocontainment settings. Importantly, the application of this system to the Ghana virus revealed that its known sequence is incomplete; however, substituting the missing sequences with those from other henipaviruses allowed us to overcome this challenge. We demonstrate that the Ghana virus replicative machinery is functional and can identify two orally efficacious antivirals effective against it. Our research offers a versatile system for life-cycle modeling of highly pathogenic henipaviruses at low biocontainment.

Supramolecular Bioconjugation Strategy for Antibody-Targeted Delivery of siRNA.

RNA interference is a widely used biological process by which double-stranded RNA induces sequence-specific gene silencing by targeting mRNA for degradation. However, the physicochemical properties of siRNAs make their delivery extremely challenging, thus limiting their bioavailability at the target site. In this context, we developed a versatile and selective siRNA delivery system of a trastuzumab-conjugated nanocarrier. These immunoconjugates consist of the assembly by electrostatic interactions of an oligonucleotide-modified antibody with a cationic micelle for the targeted delivery of siRNA in HER2-overexpressing cancer cells. Results show that, when associated with the corresponding siRNA at the appropriate N/P ratio, our supramolecular assembly was able to efficiently induce luciferase and PLK-1 gene silencing in a cell-selective manner in vitro.

Highly stable Saccharomyces cerevisiae L-BC capsids with versatile packing potential.

Virus-like particles (VLPs) are promising nanoscaffolds in development of vaccines and nanodelivery systems. Along with efficient production in various expression systems, they also offer extensive functionalization options. Nevertheless, the ultimate integrity of VLPs is an important burden for the applicability in nanobiotechnology. In this study, we characterize the Saccharomyces cerevisiae L-BC VLPs synthesized and purified from Escherichia coli and Saccharomyces cerevisiae cells. The particles exhibited prominent size stability in buffers within a range of ionic strength conditions, pH environment and presence of magnesium ions during the long-term storage at temperatures up to 37°C. Bacteria-derived particles exhibited alleviated stability in acidic pH values, higher ionic strength and temperature compared to yeast-derived particles. Taking advantage of gene engineering, 120 copies of red fluorescent protein mCherry were successfully encapsulated into both preparations of L-BC VLPs, while passive diffusion enabled encapsulation of antimicrobial peptide nisin into the yeast-derived unmodified VLPs. Our findings indicate that L-BC VLPs generally exhibit high long-term stability under various conditions, while yeast-derived L-BC VLPs are more stable under the elevated temperatures than bacteria-derived particles. Stability studies and encapsulation of particles by different molecules involving alternative strategies delineate the L-BC VLP potential to be developed into versatile nanodelivery system.

Nonviral genetic manipulation system with microenvironmental cascade sensitivity for the treatment of TNBC-associated metastatic bone tumor

Gene therapy is an efficacious therapeutic modality for triple negative breast cancer (TNBC). However, the practicable applicability is strictly confined in TNBC-associated metastatic bone tumor due to the low blood flow and poor accumulation in bone tissue, as well as the vicious cycle of mutual promotion between tumor growth and bone erosion. To address those problems, a versatile nonviral genetic manipulation system PSMCA/miR-34a, with caveolae-activated endocytosis and microenvironmental cascade-responsive release profile was exploited. In vitro and in vivo experiments confirmed that PSMCA/miR-34a not only possessed bone-targeting properties to accumulate at bone tumor tissue, also could respond to the bone tumor microacidic environment to release alendronate to inhibit the bone erosion. After internalized into cells mainly through caveolae-mediated endocytosis, it was spurred to disassemble for controlling release the miR-34a by high intracellular GSH environment, generating an effective anti-tumor effect. Moreover, due to the synergistic effect, PSMCA/miR-34a could breakdown the vicious cycle between bone destruction and tumor growth in the tumor microenvironment, showing high potential in the clinical gene therapy for TNBC-associated metastatic bone tumor.

Fabrication of a novel macrophage-targeted biomimetic delivery composite hydrogel with multiple-sensitive properties for tri-modal combination therapy of rheumatoid arthritis

In this study, a porous polydopamine (PDA) nanoparticle-decorated β-glucan microcapsules (GMs) nanoplatform (PDA/GMs) were developed with macrophage-targeted biomimetic features and a carriers-within-carriers structure. Indocyanine green (ICG) and catalase (CAT) were subsequently co-encapsulated within the PDA/GMs to create a multifunctional nanotherapeutic agent, termed CIPGs. Furthermore, CIPGs and sinomenine (SIN) were co-loaded within a thermo-sensitive hydrogel to design an injectable delivery system, termed CIPG/SH, with potential for multi-modal therapy of rheumatoid arthritis (RA). Photothermal studies indicated that the CIPGs hold excellent photothermal conversion ability and thermal stability, as they combined the photothermal performance of both PDA and ICG. Meanwhile, the CIPGs displayed favorable oxygen self-supplying and photodynamic performance. The CIPGs showed near-infrared (NIR)-induced phototoxicity, effectively inhibiting macrophage proliferation and displaying remarkable antibacterial activity. In vitro drug release from the prepared CIPG/SH showed a controlled release pattern. Animal experiments conducted on an RA mice model confirmed that the formulated CIPG/SH exhibited significant therapeutic effects. By integrating the biological advantages, photothermal/photodynamic performance of the CIPGs, and controlled drug release performance of the thermo-sensitive hydrogels in a single delivery system, the prepared injectable CIPG/SH represents a novel versatile delivery system with great potential for multi-modal combination targeting therapy in RA.

Guest-Mediated Modulation of Photophysical Pathways in a Coronene Bisimide Cyclophane.

The properties and functions of chromophores utilized by nature are strongly affected by the environment formed by the protein structure in the cells surrounding them. This concept is transferred here to host-guest complexes with the encapsulated guests acting as an environmental stimulus. A new cyclophane host based on coronene bisimide is presented that can encapsulate a wide variety of planar guest molecules with binding constants up to (4.29 ± 0.32) × 1010 M-1 in chloroform. Depending on the properties of the chosen guest, the excited state deactivation of the coronene bisimide chromophore can be tuned by the formation of host-guest complexes toward fluorescence, exciplex formation, charge separation, room-temperature phosphorescence (RTP), or thermally activated delayed fluorescence (TADF). The photophysical processes were investigated by UV/vis absorption, emission, and femto- and nanosecond transient absorption spectroscopy. To enhance the TADF, two different strategies were used by employing suitable guests: the reduction of the singlet-triplet gap by exciplex formation and the external heavy atom effect. Altogether, by using supramolecular host-guest complexation, a versatile multimodal chromophore system is achieved with the coronene bisimide cyclophane.

Self-Assembly of Silole-Based Aggregation-Induced Emission Compounds with Green Fluorescent Protein under Physiological Conditions for Traceable and Versatile Drug Delivery.

Biomacromolecules are viewed as promising drugs due to their specific functions in biological processes, biocompatibility, and pharmacological efficacy. Injective administration, chosen to avoid intestinal barriers, may in turn lead to immediate decay in the circulation system, unreliable targeting performance, or the induction of immune responses. For some biomacromolecules, chemically modified proteins have been developed for practical use. Various cargo or carrier systems are under development but have been delayed by technical difficulties. We present self-assembled nanocapsules with diameters ranging from 100 to 500 nm that can be deployed in physiological buffers to enclose various substances present in the buffers at the same time. Our amphiphilic nanocapsule, consisting of silole-core dendrimer products as the hydrophobic part and green fluorescent protein (GFP) derivatives as the hydrophilic part, connects and assembles spontaneously when mixed in solutions while engulfing dissolved or dispersed compounds together in a dose-dependent manner and shows unique optical characteristics because the dendrimer products exhibit aggregation-induced emission. Furthermore, the emission of the dendrimer causes considerable fluorescence resonance energy transfer (FRET) to GFP derivatives upon association. We could easily monitor assemblies by FRET states and particle sizes and have confirmed a stable presence in the buffer for at least a month. Further tracking of nanocapsules by fluorescence confirmed efficient uptake into some cancer cells. Nanocapsules based on GFP variants with or without a cell-surface-specific tag demonstrated that the tag improved the potential for specific targeted delivery. There were also indications that the nanocapsules became unstable after cellular uptake in the intracellular environment. We report here the simple preparation of traceable, stable, and biocompatible self-assembled nanocapsules as the basis for a versatile drug delivery system.

One-Pot Synthesis of a pH-Sensitive MOF Integrated with Glucose Oxidase for Amplified Tumor Photodynamic/Photothermal Therapy.

Photothermal therapy (PTT) and photodynamic therapy (PDT) provide targeted approaches to cancer treatment, but each therapy has inherent limitations such as insufficient tissue penetration, uneven heat distribution, extreme hypoxia, and overexpressed HSP90 in tumor cells. To address these issues, herein, by encapsulating the IR780 dye and glucose oxidase (GOx) enzyme within ZIF-8 nanoparticles, we created a versatile system capable of combining photodynamic and enhanced photothermal therapy. The integration of the IR780 dye facilitated the generation of reactive oxygen species and hyperthermia upon light activation, enabling dual-mode cancer cell ablation. Moreover, GOx catalyzes the decomposition of glucose into gluconic acid and hydrogen peroxide, leading to the inhibition of ATP production and downregulation of heat shock protein 90 (HSP90) expression, sensitizing cancer cells to heat-induced cytotoxicity. This synergistic combination resulted in significantly improved therapeutic outcomes. Both in vitro and in vivo results validated that the nanoplatform demonstrated superior specificity and favorable therapeutic responses. Our innovative approach represents a promising strategy for overcoming current limitations in cancer treatments and offers the potential for clinical translation in the future.

Functional Linkers Support Targeting of Multivalent Tweezers to Taspase1.

Taspase 1 is a unique protease not only pivotal for embryonic development but also implicated in leukemias and solid tumors. As such, this enzyme is a promising while still challenging therapeutic target, and with its protein structure featuring a flexible loop preceding the active site a versatile model system for drug development. Supramolecular ligands provide a promising complementary approach to traditional small-molecule inhibitors. Recently, the multivalent arrangement of molecular tweezers allowed the successful targeting of Taspase 1's surface loop. With this study we now want to take the next logic step und utilize functional linker systems that not only allow the implementation of novel properties but also engage in protein surface binding. Consequently, we chose two different linker types differing from the original divalent assembly: a backbone with aggregation-induced emission (AIE) properties to enable monitoring of binding and a calix[4]arene scaffold initially pre-positioning the supramolecular binding units. With a series of four AIE-equipped ligands with stepwise increased valency we demonstrated that the functionalized AIE linkers approach ligand binding affinities in the nanomolar range and allow efficient proteolytic inhibition of Taspase 1. Moreover, implementation of the calix[4]arene backbone further enhanced the ligands' inhibitory potential, pointing to a specific linker contribution.

A modular and flexible open source cell incubator system for mobile and stationary use

Culturing living cells in vitro requires the maintenance of physiological conditions for extended periods of time. Here, we introduce a versatile and affordable incubation system, addressing the limitations of traditional incubation systems. Conventionally, stationary cell incubators maintain constant temperature and gas levels for in vitro cell culturing. Combining such incubators with additional lab equipment proves challenging. The presented platform offers modularity and adaptability, enabling customization to diverse experimental needs. The system includes a main unit with a user-friendly interface as well as an interchangeable incubation chamber. We present two incubation chambers targeting two completely different use cases. The first chamber, named “inkugo” facilitates the transportation of cells for up to two hours without external power and for more than a day without an external CO2 source. The second chamber termed “inkubox” was designed to enable continuous electrophysiological recordings. Recordings from up to four neural cultures growing on high-density microelectrode arrays can be performed in parallel. The system’s unique feature lies in its separability of control and incubation components, allowing one control unit to manage various custom chambers. The design’s simplicity and the use of widely accessible components make the here proposed incubation system replicable for any laboratory. This platform fosters collaboration and experimentation in both decentralized and traditional laboratory settings, making it an invaluable addition to any cell culturing pipeline.

A compact, versatile drug-induced splicing switch system with minimal background expression

Gene-switch techniques hold promising applications in contemporary genetics research, particularly in disease treatment and genetic engineering. Here, we developed a compact drug-induced splicing system that maintains low background using a human ubiquitin C (hUBC) promoter and optimized drug (LMI070) binding sequences based on the Xon switch system. To ensure precise subcellular localization of the protein of interest (POI), we inserted a 2A self-cleaving peptide between the extra N-terminal peptide and POI. This streamlined and optimized switch system, named miniXon2G, effectively regulated POIs in different subcellular localizations both invitro and invivo. Furthermore, miniXon2G could be integrated into endogenous gene loci, resulting in precise, reversible regulation of target genes by both endogenous regulators and drugs. Overall, these findings highlight the performance of miniXon2G in controlling protein expression with great potential for general applicability to diverse biological scenarios requiring precise and delicate regulation.

A novel framework for production planning and class-based storage location assignment: Multi-criteria classification approach

Efficient warehouse management is essential for optimizing inventory, minimizing transportation costs, and enhancing overall performance. This research introduces a novel Mixed-Integer Nonlinear Programming (MINLP) model to address the Storage Location Assignment Problem (SLAP) in warehouse management. Integrating multi-criteria decision-making with strategic production planning, our model advances warehouse operations by allocating storage locations to products strategically, focusing on reducing transportation distances and maximizing storage efficiency. The distinctive innovation of this study is the nuanced application of the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) results to strategic storage location assignments, enhancing the model's capability to consider a comprehensive evaluation of inventory attributes, including physical characteristics and perishability. This approach evolves TOPSIS's application in warehouse management, enabling it to consider both physical characteristics and perishability of products. The outcomes of TOPSIS, including product classifications and preferences, serve as vital inputs to the mathematical model, facilitating a comprehensive evaluation of storage locations that encompasses spatial, demand-related, and physical aspects of inventory. Additionally, our research introduces a versatile decision support system, adaptable to various operational requirements. This system enhances practical decision-making in warehouse management, accommodating scenarios based on single or multiple criteria, including the cube-per-order index (COI). The research results highlight the significant impact of this innovative approach in enhancing warehouse management. By addressing the complexities of storage location assignment and integrating multiple criteria, we achieve more efficient and cost-effective warehouse operations. The approach has been shown to be adaptable and practical, making it a valuable contribution to the field of logistics and warehouse management.

Home Automation System using OpenCV and Arduino

The research focuses on the integration of computer vision capabilities through OpenCV with Arduino-based hardware to create an efficient and versatile home automation system. The system employs cameras for image processing, allowing it to recognize and respond to various environmental changes within the home. The hardware setup utilizes Arduino microcontrollers as the central processing units, enabling seamless communication between the OpenCV algorithms and the connected devices. A user-friendly interface facilitates remote monitoring and control of home appliances through a mobile or web application. By combining the power of OpenCV for image processing and Arduino for real-time control, this home automation system provides a smart and adaptive solution for enhancing security, energy efficiency, and overall convenience within a household. The research’s modular design allows for future expansions and integration with emerging technologies, ensuring scalability and relevance in the rapidly evolving field of smart home automation.

A versatile system for the growth of porphyrin films via electrospray and molecular sublimation in vacuum and their multi-technique characterization.

We present a system for the growth of molecular films in vacuum that exhibits high versatility with respect to the choice of molecular species. These can be either evaporated from powders or injected from solutions using an electrospray system, making it possible to handle particularly large and/or fragile molecules in a controlled environment. The apparatus is equipped with a reflectance anisotropy spectroscopy system for the in situ characterization of the optical response of the films and can be directly connected to a photoelectron spectrometer without breaking the vacuum. The system is conceived for the study and characterization of porphyrin films. Here, to showcase the range of possible analyses allowed by the experimental setup and test the operation of the system, novel results are provided on electrospray deposition on highly oriented pyrolytic graphite of Zn tetraphenyl porphyrins and Zn proto porphyrins, the latter featuring fragile side groups that make deposition from solution more attractive. In situ characterization is complemented by ex situ atomic force microscopy. Thanks to this multi-technique approach, changes in the film morphology and spectroscopic response are detected and directly related to the choice of the molecular moiety and growth method.

Detection of Intrusions using Transformer - LSTM Model

The project focuses on addressing the escalating issue of web attacks and ensuring cyberspace security in the era of advancing internet technology. The primary goal is to enhance intrusion detection systems (IDS) by moving beyond traditional keyword and rule- based methods. The project explores the integration of artificial intelligence, specifically deep learning techniques, to develop a more robust and adaptive approach to identifying network intrusion events. With the limitations of conventional detection methods in mind, there is a critical need for a more effective and versatile intrusion detection system. The rise in diverse attack methods necessitates an intelligent system capable of autonomously adapting to new attack patterns without relying on manual rule creation. This project is designed to benefit both individuals and organizations reliant on network services. Improved intrusion detection ensures a higher level of cybersecurity, safeguarding sensitive data and systems. The project's outcomes have the potential to contribute significantly to the broader field of network security. Leveraging the power of deep learning, the proposed model- Transformer (autoencoder) LSTM combines Transformer and LSTM architectures. Multiple attention mechanisms are employed to select and extract features, while the LSTM model facilitates understanding the sequential relationships within network traffic, enabling the accurate identification of intrusion events. And we are comparing the model with DNN, LSTM and Transformer(autoencoder). To enhance performance, CNN and CNN+LSTM extensions were incorporated into the project, complementing the Transformer (autoencoder) LSTM model. These models aim to further refine feature extraction and exploit spatial relationships within network traffic. Key Words:intrusion detection, transformer, LSTM, deep learning.

Classical Monocytes Shuttling for Precise Delivery of Nanotherapeutics to Glioblastoma.

Glioblastoma (GBM) is the most aggressive brain tumor for which current therapies have limited efficacy. Immunosuppression and difficulties in accessing tumors with therapeutic agents are major obstacles for GBM treatments. Classical monocytes (CMs) possess the strongest infiltration among myeloid cells recruited into tumors during tumorigenesis. In this study, CMs are utilized to deliver the small-molecule CUDC-907 encapsulated in nanoparticles (907-NPs@CMs) for GBM therapy. Hitchhiking on CMs enables more 907-NPs to successfully penetrate the blood-brain barrier (BBB) and reach the interior of tumors. Results demonstrate that 907-NPs@CMs significantly improve the survival rates by suppressing tumor growth and reversing the immunosuppression of tumor microenvironment (TME). Furthermore, the high delivery efficiency of CMs reduces the amount of CUDC-907 required for treatments, reducing the physiological toxicity and off-target effects caused by high doses. 907-NPs@CMs is a safe and versatile therapeutic system that provides a platform for targeted drug delivery to tumors and the ability to treat GBM through a combination of chemotherapy and immunotherapy.

Performance analysis of a versatile bulk-service queue with group-arrival, batch-size-dependent service time and queue-length-dependent vacation

ABSTRACT Queueing systems with bulk-service and vacation policy have become one of the pivotal interest for the researchers due to their widespread application in food processing technology, manufacturing systems, power consumption in small cell base stations etc. This article analyzes a single server versatile bulk-service queueing system wherein the customers arrive according to a compound Poisson process and the service time is dependent on the batch size of undergoing service. Moreover, single and multiple vacation policies have been incorporated along with queue-length-dependent vacation. After providing the steady-state system equations, the bivariate probability generating function of queue and server content distribution together has been derived at departure epoch. After the evaluation of the unknown probabilities, complete joint distributions have been extracted in terms of roots of the denominator of the bivariate probability generating function. The discussed procedure and the reported results have been depicted through some numerical examples for different service and vacation time distributions. Some significant observation about the model has been sketched graphically.

Chaos-Assisted Production of Micro-Architected Spheres (CAPAS).

Hydrogel droplets with inner compartments are valuable in various fields, including tissue engineering. A droplet-based biofabrication method is presented for the chaos-assisted production of architected spheres (CAPAS) for the rapid generation of multilayered hydrogel spheres (ranging from 0.6 to 3.5mm in diameter) at high-throughput rates (up to 2000 spheres per min). This method is based on the use of chaotic advection generated by a Kenics static mixer (KSM) nozzle. The configuration of the KSM (i.e., the number of mixing elements) determines the number of compartments within the sphere. Sphere size is adjusted by flow rate, printhead outlet diameter, polymer concentration (sodium alginate or gelatin-methacryloyl (GelMA)), and crosslinking bath composition. This versatile system operates in dripping and jetting modes, preserving multilayered architecture in both modes. Proof-of-concept experiments with breast cancer (MDA-MB-231), human dermal fibroblast (HDF), and murine myoblast (C2C12) lines show over 80% cell viability immediately post-fabrication, maintained over extended culture (14 or 30 days). CAPAS is used to create a breast cancer model with cancer-tissue-like and healthy-tissue-like micro-niches to test paclitaxel doses. It is envisioned that CAPAS will enable high-throughput fabrication of hydrogel spheres for tissue engineering, chemical engineering, and material sciences applications.

At the Limit of Interfacial Sharpness in Nanowire Axial Heterostructures.

As semiconductor devices approach dimensions at the atomic scale, controlling the compositional grading across heterointerfaces becomes paramount. Particularly in nanowire axial heterostructures, which are promising for a broad spectrum of nanotechnology applications, the achievement of sharp heterointerfaces has been challenging owing to peculiarities of the commonly used vapor-liquid-solid growth mode. Here, the grading of Al across GaAs/AlxGa1-xAs/GaAs heterostructures in self-catalyzed nanowires is studied, aiming at finding the limits of the interfacial sharpness for this technologically versatile material system. A pulsed growth mode ensures precise control of the growth mechanisms even at low temperatures, while a semiempirical thermodynamic model is derived to fit the experimental Al-content profiles and quantitatively describe the dependences of the interfacial sharpness on the growth temperature, the nanowire radius, and the Al content. Finally, symmetrical Al profiles with interfacial widths of 2-3 atomic planes, at the limit of the measurement accuracy, are obtained, outperforming even equivalent thin-film heterostructures. The proposed method enables the development of advanced heterostructure schemes for a more effective utilization of the nanowire platform; moreover, it is considered expandable to other material systems and nanostructure types.

Validation of inertial measurement units based on waveform similarity assessment against a photogrammetry system for gait kinematic analysis.

When assessing gait analysis outcomes for clinical use, it is indispensable to use an accurate system ensuring a minimal measurement error. Inertial Measurement Units (IMUs) are a versatile motion capture system to evaluate gait kinematics during out-of-lab activities and technology-assisted rehabilitation therapies. However, IMUs are susceptible to distortions, offset and drifting. Therefore, it is important to have a validated instrumentation and recording protocol to ensure the reliability of the measurements, to differentiate therapy effects from system-induced errors. A protocol was carried out to validate the accuracy of gait kinematic assessment with IMUs based on the similarity of the waveform of concurrent signals captured by this system and by a photogrammetry reference system. A gait database of 32 healthy subjects was registered synchronously with both devices. The validation process involved two steps: 1) a preliminary similarity assessment using the Pearson correlation coefficient, and 2) a similarity assessment in terms of correlation, displacement and gain by estimating the offset between signals, the difference between the registered range of motion (∆ROM), the root mean square error (RMSE) and the interprotocol coefficient of multiple correlation (CMCP). Besides, the CMCP was recomputed after removing the offset between signals (CMCPoff). The correlation was strong (r > 0.75) for both limbs for hip flexion/extension, hip adduction/abduction, knee flexion/extension and ankle dorsal/plantar flexion. These joint movements were studied in the second part of the analysis. The ∆ROM values obtained were smaller than 6°, being negligible relative to the minimally clinically important difference (MCID) estimated for unaffected limbs, and the RMSE values were under 10°. The offset for hips and ankles in the sagittal plane reached -9° and -8°, respectively, whereas hips adduction/abduction and knees flexion/extension were around 1°. According to the CMCP, the kinematic pattern of hip flexion/extension (CMCP > 0.90) and adduction/abduction (CMCP > 0.75), knee flexion/extension (CMCP > 0.95) and ankle dorsi/plantar flexion (CMCP > 0.90) were equivalent when captured by each system synchronously. However, after offset correction, only hip flexion/extension (CMCPoff = 1), hip adduction/abduction (CMCPoff > 0.85) and knee flexion/extension (CMCPoff > 0.95) satisfied the conditions to be considered similar.