Cargo Loading Research Articles

RNA and Protein Delivery by Cell-Secreted and Bioengineered Extracellular Vesicles.

Extracellular vesicles (EVs) are carriers of biological signals through export and delivery of RNAs and proteins. Of increasing interest is the use of EVs as a platform for delivery of biomolecules. Preclinical studies have effectively used EVs to treat a number of diseases. Uniquely, endogenous machinery within cells can be manipulated in order to produce desirable loading of cargo within secreted EVs. In order to inform the development of such approaches, an understanding of the cellular mechanisms by which cargo is sorted to EVs is required. Here, the current knowledge of cargo sorting within EVs is reviewed. Here is given an overview of recent bioengineering approaches that leverage these advances. Methods of externally manipulating EV cargo are also discussed. Finally, a perspective on the current challenges of EVs as a drug delivery platform is offered. It is proposed that standardized bioengineering methods for therapeutic EV preparation will be required to create a well-defined clinical product.

Electrical Propulsion and Cargo Transport of Microbowl Shaped Janus Particles.

Herein the effective electrical propulsion, cargo trapping, and transport capabilities of microbowl-shaped Janus particles (JPs) are demonstrated and evaluated. These active JPs are made by deposition of Au and Ti layers onto sacrificial spherical polystyrene particles, followed by oxidation of the Ti to TiO2 . In contrast to the commonly studied spherical JP, the dual broken symmetry of both geometrical and electrical properties of the microbowl renders a strong dependence of its mobility and cargo loading on the order of the layering of Au and TiO2 . Specifically, an opposite direction of motion is obtained for interchanged layers of Au and TiO2 , using only electrical propulsion as the sole mechanism of motion. The concave side of the microbowl exhibits a negative dielectrophoretic trap of large size wherein trapped cargo is protected from hydrodynamic shearing, leading to an enhanced cargo loading capacity compared to that obtained using common spherical JP. Such enhanced cargo capability of the microbowl along with the ease of engineering it by interchanging the order of the layers are very attractive for future in vitro biological and biomedical applications.

Bone Regeneration Using MMP-Cleavable Peptides-Based Hydrogels.

Accumulating evidence has suggested the significant potential of chemically modified hydrogels in bone regeneration. Despite the progress of bioactive hydrogels with different materials, structures and loading cargoes, the desires from clinical applications have not been fully validated. Multiple biological behaviors are orchestrated precisely during the bone regeneration process, including bone marrow mesenchymal stem cells (BMSCs) recruitment, osteogenic differentiation, matrix calcification and well-organized remodeling. Since matrix metalloproteinases play critical roles in such bone metabolism processes as BMSC commitment, osteoblast survival, osteoclast activation matrix calcification and microstructure remodeling, matrix metalloproteinase (MMP) cleavable peptides-based hydrogels could respond to various MMP levels and, thus, accelerate bone regeneration. In this review, we focused on the MMP-cleavable peptides, polymers, functional modification and crosslinked reactions. Applications, perspectives and limitations of MMP-cleavable peptides-based hydrogels for bone regeneration were then discussed.

DEM modeling of the effects of external loads and material properties on the stability of nickel ore in a cargo hold

Cargo liquefaction leading to the serious loss of ship stability is attracting more attention from the international community. However, due to the complexity of external loads and the properties of the cargo, the mechanism of cargo liquefaction has not been revealed for every situation. In this study, considering the effects of the moisture content, particle size grading, initial porosity and motions of a typical cargo hold, a three-dimensional full-scale numerical model based on the discrete element method is established, and numerical investigations are carried out. To better ensure that the full-scale model conditions are consistent with the reality and the rationality of particle amplification, a static incline test and a cargo loading test are also simulated and verified. The results show that the effect of the moisture content on the cargo stability can be estimated using the friction coefficients between particles. Additionally, the moisture content is the key factor that affects the stability of the cargo and promotes its liquefaction. Furthermore, there are critical values for fine particle content, at which cargo fluidity is greatly enhanced. For external loads, the cargo stability is greatly weakened with the increase of the motion amplitude. Furthermore, there is a critical frequency, beyond which the cargo stability will be further damaged.

Experimental assessment of eigenfrequencies and stiffness of the elastically supported body

The aim of the research is to derive an analytical solution describing the vibrations of railway wagons caused by asymmetrical loading of cargo for prediction of dangerous conditions. A presumption for the dynamic analysis and assessment of the driving quality of railway vehicles is an important experimental determination and verification of the basic characteristics of the suspension respectively. A method for determining these characteristics for two-axle vehicles has been developed and applied. This method requires knowledge of the position of the center of gravity and the main central moments of inertia. Vehicle bodies, chassis frames, and wheelsets are assumed to be rigid bodies under the investigation of these characteristics. The characteristics of the springs are linear or can be linearized. The vehicle body performs a general spatial movement. The result of the investigation is an analytical method to obtain the required characteristics needed for the dynamic analysis of the vehicle.

Orodispersible films: Conception to quality by design.

Orodispersible films (ODFs) are ultra-thin, stamp-sized, elegant, portable and patient-centric pharmaceutical dosage forms that do not need water to be ingested. They are particularly useful for paediatric and geriatric patient populations with special needs such as dysphagia, Parkinson's disease, and oral cancer. Accordingly, they hold tremendous potential in gaining patient compliance, convenience and pharmacotherapy. In the present review, conception and evolution of ODFs as a product and its technology are discussed. The review continues by providing overview about the potential of ODFs as carriers for delivering drugs, herbal extracts, probiotics and vaccines. Besides, strategies employed in drug cargo loading, taste masking of bitter drugs and enhancing drug stability are discussed. Finally, the review concludes by providing a brief overview about quality by design (QbD) principles in development of ODFs.

Shipping optimization systems (SOS) for tramp: stochastic cargo soft time windows

Cargo time windows have been discussed in the shipping literature in several ways and within several contexts. One way considers the hard time windows, where a fixed date is assigned to both the open and close laycan for the cargo loading and discharging ports. According to this way, the cargo is rejected if the ship cannot meet these time windows. Another way develops soft time windows, where the ship for an additional cost or a penalty may violate the cargo laycan. The context in the previous ways includes a tramp deterministic model. In this paper, the soft time windows are considered within a context of a tramp stochastic model, where both the cargo transport demand and the laycan dates are random variables. The objective is a gross-profit-per-day accompanied with realistic shipping elements embedded into decision support systems known as shipping optimization systems (SOS). The chance-constraint programming and the modified version of the Dantzig–Wolfe decomposition principle known as block-angular linear ratio programming solve the problem. A case study demonstrates that using stochastic soft time windows with stochastic cargo transport demand has considerably improved the gross profit-per-day better than trying to arrive on time by varying ship speed or being within a non-stochastic shipping context.

Engineering Cardiac Small Extracellular Vesicle-Derived Vehicles with Thin-Film Hydration for Customized microRNA Loading.

Cell therapies for myocardial infarction, including cardiac ckit+ progenitor cell (CPC) therapies, have been promising, with clinical trials underway. Recently, paracrine signaling, specifically through small extracellular vesicle (sEV) release, was implicated in cell-based cardiac repair. sEVs carry cardioprotective cargo, including microRNA (miRNA), within a complex membrane and improve cardiac outcomes similar to that of their parent cells. However, miRNA loading efficiency is low, and sEV yield and cargo composition vary with parent cell conditions, minimizing sEV potency. Synthetic mimics allow for cargo-loading control but consist of much simpler membranes, often suffering from high immunogenicity and poor stability. Here, we aim to combine the benefits of sEVs and synthetic mimics to develop sEV-like vesicles (ELVs) with customized cargo loading. We developed a modified thin-film hydration (TFH) mechanism to engineer ELVs from CPC-derived sEVs with pro-angiogenic miR-126 encapsulated. Characterization shows miR-126+ ELVs are similar in size and structure to sEVs. Upon administration to cardiac endothelial cells (CECs), ELV uptake is similar to sEVs too. Further, when functionally validated with a CEC tube formation assay, ELVs significantly improve tube formation parameters compared to sEVs. This study shows TFH-ELVs synthesized from sEVs allow for select miRNA loading and can improve in vitro cardiac outcomes.

Feasibility Study on the Imperatives for the Simulation of Ammonia Release Management

Ammonia is being exported through Bintulu Port via pipeline and loaded to the vessel (ship) using marine loading arm. Ammonia is toxic in nature and transferred at -33 oC. Marine loading arms are special equipment for loading and unloading liquid cargo from the wharf and the vessel with swivel joints, and supplemented by supporting structure, and other accessories. Loading arms have safety features such as that we can set up quick release mechanism, in which the loading arm will decouple from the manifold when there is an emergency, or the movement of vessel is out of range. Studies such as by United Kingdom (UK) Health and Safety Executive (HSE) on loading arm had provided the probabilities of failures, and the possible size of hole such as guillotine break or 0.1 cross sectional area of pipe. This information, with ammonia operational parameters such as internal and external pressure and ammonia liquid flow rate we can thus predict the amount of ammonia release. Combining with meteorological data information, local landscape conditions and utilizing ALOHA software, we are able to simulate ammonia dispersion and thus predicting the impact of toxicity of ammonia release on the population within the area. This paper is a feasibility study that use modelling approaches to manage ammonia leakage.

Triggered Reversible Disassembly of an Engineered Protein Nanocage*.

Protein nanocages play crucial roles in sub-cellular compartmentalization and spatial control in all domains of life and have been used as biomolecular tools for applications in biocatalysis, drug delivery, and bionanotechnology. The ability to control their assembly state under physiological conditions would further expand their practical utility. To gain such control, we introduced a peptide capable of triggering conformational change at a key structural position in the largest known encapsulin nanocompartment. We report the structure of the resulting engineered nanocage and demonstrate its ability to disassemble and reassemble on demand under physiological conditions. We demonstrate its capacity for in vivo encapsulation of proteins of choice while also demonstrating in vitro cargo loading capabilities. Our results represent a functionally robust addition to the nanocage toolbox and a novel approach for controlling protein nanocage disassembly and reassembly under mild conditions.

Triggered Reversible Disassembly of an Engineered Protein Nanocage**

AbstractProtein nanocages play crucial roles in sub‐cellular compartmentalization and spatial control in all domains of life and have been used as biomolecular tools for applications in biocatalysis, drug delivery, and bionanotechnology. The ability to control their assembly state under physiological conditions would further expand their practical utility. To gain such control, we introduced a peptide capable of triggering conformational change at a key structural position in the largest known encapsulin nanocompartment. We report the structure of the resulting engineered nanocage and demonstrate its ability to disassemble and reassemble on demand under physiological conditions. We demonstrate its capacity for in vivo encapsulation of proteins of choice while also demonstrating in vitro cargo loading capabilities. Our results represent a functionally robust addition to the nanocage toolbox and a novel approach for controlling protein nanocage disassembly and reassembly under mild conditions.

Extracellular vesicles derived from physiologically loaded living myocardial slices improve the contractility of healthy living myocardial slices

Abstract Introduction Mechanical load is a determinant of the healthy-to-failing myocardium transition. Extracellular vesicles (EVs) found in the range of 30–200 nm contain bioactive cargoes such as DNA, miRNA and proteins, and thus are considered important mediators of the paracrine crosstalk between cells in health and disease. Living myocardial slices (LMS) are 300μm layers of myocardial tissue that allow modelling of the intercellular communication in the heart. Purpose To assess whether secretion of EVs derived from physiological or pathological mechanically-loaded LMS regulate changes in myocardial contractility. Methods LMS were prepared from adult male Sprague-Dawley rats' left ventricle using a vibratome. LMS were stretched uniaxially using customised stretchers at a sarcomere length of 2.2μm and 2.4μm to recapitulate a physiological (P) and an overload (O) condition, respectively. LMS were cultured for 48-hours under electromechanical stimulation in M-199 media with Earls' salts, catecholamines, dexamethasone and triiodothyronine. After culture, media was harvested and processed for exosomes isolation by size exclusion chromatography. Particle size and concentration was assessed by nanoparticle tracking analysis. To validate the different phenotypes in LMS, force-stretch relationship was assessed after 48-hours culture. EVs isolated from P and O LMS were then used to treat physiologically loaded LMS (1x107 particles/LMS) and the force-stretch relationship was assessed after 48-hours culture. Results No significant difference in contractility was found between P and O LMS at any stretch increment (5.99±1.10 vs 2.82±0.92 mN/mm2 max force), whilst a significant increase in the passive tension of myocardial tissue was observed at 25% (15.90±2.77 vs 4.67±0.37 mN/mm2; p<0.05,) and 30% stretch (21.92±3.60 vs 5.73±0.6 mN/mm2; p<0.001) in the P LMS compared to O LMS (n=11 P, n=4 O). EVs quantification showed no significant difference between P and O conditions (1.16329E+11±4587386608 vs 1.08915E+11±7641744894 particles/ml); (n=7 P; n=3 O). However, P EVs-treated LMS showed a significant increase in contractility after 48-hours, compared to PBS-treated LMS at 25% (8.49±0.62 vs 5.34±0.9 mN/mm2) and 30% stretch (9.55±0.42 vs 5.99±1.10 mN/mm2; p<0.05); (n=11 PBS, n=4 P EVs,) and O EVs-treated LMS from 15–30% stretch (9.55±0.42 vs 3.23±0.55 mN/mm2 max force; p<0.0001); (n=4 P EVs, n=5 O EVs). Conversely, no significant differences in contractility were found between PBS-treated LMS and O EVs-treated LMS (5.99±1.10 vs 3.23±0.55 mN/mm2 maximum force); (n=5 O EVs, n=11 PBS). Conclusions EVs derived from physiologically loaded LMS improve the contractility of healthy LMS after 48-hours culture compared to PBS-treated LMS. Understanding how mechanical load correlates with specific loading of cargoes in EVs may reveal interesting therapeutic targets for the treatment of heart failure. Funding Acknowledgement Type of funding sources: Private grant(s) and/or Sponsorship. Main funding source(s): British Heart Foundation Physiological and overloaded phenotypesContractility of EVs treated LMS

Self-Assembled Nanosized Vehicles from Amino Acid-Based Amphiphilic Polymers with Pendent Carboxyl Groups for Efficient Drug Delivery.

Developing safe and efficient delivery vehicles for chemotherapeutic drugs has been a long-standing demanding. Amino acid-based polymers are promising candidates to address this challenge due to their excellent biocompatibility and biodegradation. Herein, a series of well-defined amphiphilic block copolymers were prepared by PET-RAFT polymerization of N-acryloyl amino acid monomers. By altering monomer types and the block ratio of the copolymers, the copolymers self-assembled into nanostructures with various morphologies, including spheres, rod-like, fibers, and lamellae via hydrophobic and hydrogen bonding interactions. Significantly, the nanoparticles (NPs) assembled from amphiphilic block copolymers poly(N-acryloyl-valine)-b-poly(N-acryloyl-aspartic acid) (PV-b-PD) displayed an appealing cargo loading efficiency (21.8-32.6%) for a broad range of drugs (paclitaxel, doxorubicin (DOX), cisplatin, etc.) due to strong interactions. The DOX-loaded PV-b-PD NPs exhibited rapid cellular uptake (within 1 min) and a great therapeutic performance. These drug delivery systems provide new insights for regulating the controlled morphologies and improving the efficiency of drug delivery.

A simulation approach for aircraft cargo loading considering weight and balance constraints

Air cargo transport is a growing industry in parallel with the growth in world trade and e-commerce. The global air cargo transport traffic getting busier, the importance of timely loading with minimum error is increasing. Therefore, digitalization of the air cargo loading process is needed. Assignment of Unit Load Devices (ULDs) to the specific positions on the freighter is performed by loadmasters by manual or semi-manual methods. This study aims to design a simulation model, which performs the ULD assignment automatically by simulating the loading process performed by the experienced loadmasters under the weight and balance constraints. The SEMMA (sample, explore, modify, model, assess) model is used while generating the simulation model. Fifty real-world loading orders were used to assess the performance of the model. The ULD assignment process by the simulation model and the loadmasters using semi-manual loading were compared with regard to time and center of gravity performance indicators. The results demonstrated that the simulation model can load all the given sets of ULDs, as efficiently as a loadmaster with a similar center of gravity in a shorter period of time. In conclusion, the proposed simulation model provides an efficient solution to the ULD assignment problem. However, the base model generated may be improved to address various real-world scenarios

Nanomaterial Strategies for Delivery of Therapeutic Cargoes

AbstractOver the last decade, much progress has been made in developing nanoparticle‐mediated delivery systems to overcome the limitations of existing in vivo delivery technologies. However, the balance between efficacy and safety continues to limit the clinical translation of nanoscale delivery systems. Furthermore, optimizing delivery efficiency requires tuning nanoparticle type and attachment chemistry, both of which are dependent on the cargo being delivered. While the delivery of protein therapeutics is of particular interest, the complexity inherent to protein cargo introduces additional challenges for cargo loading and stabilization. Advances needed for efficient and safe in vivo nanoparticle delivery systems should prioritize design strategies that co‐optimize safety, biodegradability, and covalent functionalization. In this review, the most commonly used non‐viral nanoparticles are outlined for delivery of small molecule drug, nucleic acid, and protein therapeutic cargoes and potential strategies are discussed for rationally designing nanoparticle‐mediated delivery systems.

Time series forecasting for port throughput using recurrent neural network algorithm

ABSTRACT Container throughput is a critical factor to appraise a seaport performance and predicting this measure has played a vital role in port operations. Within the scope of effective decision making, predictive techniques have been presented for forecasting port throughput. Based on observing throughput variations in seasonal patterns and business cycles, the obtained data might help port authority to make better and more accurate decisions and improve seaport productivity. By applying predictive methods to the throughput data of Singapore and Busan port, the decision-makers can assess forecasting accuracy by measuring prediction errors. The numerical tests show that the echo state network (ESN) provides a high level of accuracy for predicting container throughput. As a result, the port managers could make use of this decision support strategy to foresee short-term plans for improving facilities, establishing effective cargo loading and unloading plans, consequently ensuring port productivity and profitability.

A Study on the Bonded Flush Patch Repair Method for Aircraft Composite Solid Laminates

An evaluation was performed by applying repair technology of composite solid laminates to airframe composite structures. Damage and defects in the composite laminate structure of an aircraft are caused by events occurring during vehicle movement, such as lightning, tide collisions, and runway collisions during takeoff and landing along with damage caused during aircraft movement and maintenance, fuel injection, and cargo loading. After the damaged part was removed, the damaged composite solid laminated structure was repaired using the bonded flush patch repair method. In the bonded flush patch repair method, the molded patch is joined to the base material using an adhesive. To determine the damage caused by external impact, the test results of the repaired specimen and those of the specimen before damage were compared.

Injectable Natural Polymer Hydrogels for Treatment of Knee Osteoarthritis.

Osteoarthritis (OA) is a serious chronic and degenerative disease that increasingly occurs in the aged population. Its current clinical treatments are limited to symptom relief and cannot regenerate cartilage. Although a better understanding of OA pathophysiology has been facilitating the development of novel therapeutic regimen, delivery of therapeutics to target sites with minimal invasiveness, high retention, and minimal side effects remains a challenge. Biocompatible hydrogels have been recognized to be highly promising for controlled delivery and release of therapeutics and biologics for tissue repair. In this review, the current approaches and the challenges in OA treatment, and unique properties of injectable natural polymer hydrogels as delivery system to overcome the challenges are presented. The common methods for fabrication of injectable polysaccharide-based hydrogels and the effects of their composition and properties on the OA treatment are detailed. The strategies of the use of hydrogels for loading and release cargos are also covered. Finally, recent efforts on the development of injectable polysaccharide-based hydrogels for OA treatment are highlighted, and their current limitations are discussed.

Thermal stratification and rollover phenomena in liquefied natural gas tanks

A non-equilibrium multilayer thermodynamic model is developed to predict the thermal stratification and rollover phenomena in liquefied natural gas (LNG) storage tanks. This model considers the boundary layer formation along the tank walls and uses an adaptive mesh to accommodate the changes in the LNG level in the tank over time. The accuracy of the model is verified against experiment data available in the literature to predict the thermal stratification and rollover in cryogenic storage tanks. A parametric study is conducted to investigate the key factors affecting the rollover start time. The results indicate that in an LNG storage tank at atmospheric pressure, the ratio between the amount of fresh LNG loaded to the tank (cargo) and the amount of LNG left in the tank (heel) prior to the loading of cargo, and the methane concentration in the cargo layer have direct effects on the rollover start time. The amount of heat transfer to the tank, the tank aspect ratio, and the nitrogen concentration in the heel or cargo layers affect the LNG evaporation rate. Our results indicate that the cargo to heel ratio should be determined with caution to prevent the rollover in LNG storage tanks.

Biomimetic Bacterial Membrane Vesicles for Drug Delivery Applications.

Numerous factors need to be considered to develop a nanodrug delivery system that is biocompatible, non-toxic, easy to synthesize, cost-effective, and feasible for scale up over and above their therapeutic efficacy. With regards to this, worldwide, exosomes, which are nano-sized vesicles obtained from mammalian cells, are being explored as a biomimetic drug delivery system that has superior biocompatibility and high translational capability. However, the economics of undertaking large-scale mammalian culture to derive exosomal vesicles for translation seems to be challenging and unfeasible. Recently, Bacterial Membrane Vesicles (BMVs) derived from bacteria are being explored as a viable alternative as biomimetic drug delivery systems that can be manufactured relatively easily at much lower costs at a large scale. Until now, BMVs have been investigated extensively as successful immunomodulating agents, but their capability as drug delivery systems remains to be explored in detail. In this review, the use of BMVs as suitable cargo delivery vehicles is discussed with focus on their use for in vivo treatment of cancer and bacterial infections reported thus far. Additionally, the different types of BMVs, factors affecting their synthesis and different cargo loading techniques used in BMVs are also discussed.