Disruption Methods Research Articles

An efficient marker recycling system for sequential gene deletion in a deep sea-derived fungus Acremonium sp. HDN16-126

Acremonium species are prolific producers of therapeutic molecules which include the widely used beta-lactam antibiotic, cephalosporin. In light of their significant medical value, an efficient gene disruption method is required for the physiological and biochemical studies on this genus of fungi. However, the number of selection markers that can be used for gene targeting is limited, which constrain the genetic analysis of multiple functional genes. In this study, we established a uridine auxotrophy based marker recycling system which achieves scarless gene deletion, and allows the use of the same selection marker in successive transformations in a deep sea-derived fungus Acremonium sp. HDN16-126. We identified one homologue of Acremonium chrysogenum pyrG (also as a homologous gene of the yeast URA3) from HDN16-126, designated as pyrG-A1, which can be used as a selection marker on uridine free medium. We then removed pyrG-A1 from HDN16-126 genome via homologous recombination (HR) on MM medium with 5-fluoroortic acid (5-FOA), a chemical that can be converted into a toxin of 5-flurouracil by pyrG-A1 activity, thus generating the HDN16-126-△pyrG mutant strain which showed auxotrophy for uridine but insensitivity to 5-FOA and enabled the use of exogenous pyrG gene as both positive and negative selection marker to achieve the scarless deletion of target DNA fragments. We further applied this marker recycling system to successfully disrupt two target genes pepL (encodes a putative 2OG-Fe (II) dioxygenase) and pepM (encodes a putative aldolase) identified from HDN16-126 genome, which are proposed to be functional genes related to 2-aminoisobutyric acid metabolism in fungi. This work is the first application of uridine auxotrophy based scarless gene deletion method in Acremonium species and shows promising potential in assisting sequential genetic analysis of filamentous fungi.

Life cycle assessment with parameterised inventory to derive target values for process parameters of microalgae biorefineries

This paper presents a parameterised life cycle assessment approach to determine threshold values for input streams and target process yields to lower the resource consumption and ensure environmental competitiveness of a microalgal biorefinery producing substitutes for surfactants, palm oil, bioplastic feedstock and animal feed from Chlorella vulgaris. The biorefinery concept in this work investigated four different extraction pathways. 38 input parameters were defined and used to calculate the inventory of the biorefinery. Experimental and literature data were used to determine the lower and upper limits of the input parameters. A quasi-random Saltelli sample with a size of 4000 different combinations of input parameters was generated for a global sensitivity analysis. Different sources of heat, carbon dioxide and nutrients were considered as background system as well as a renewable electricity mix and on-site energy generation. The global sensitivity analysis was followed by a reverse analysis to identify threshold values for the key parameters. Membrane technologies for extracting proteins and polysaccharides show improved results compared to chemical extractions, while for lipid recovery, supercritical carbon extraction is more promising than conventional extraction. Needs for improvement were identified for the energy demand of drying processes, the efficiency of disruption methods, which should dissolve at least 50% of the proteins with less than 5kWh electricity per kg microalgae, as well as for the protein recovery yields of membrane technologies, which should exceed 70%. The outcomes suggest that microalgal production facilities should be integrated into existing industries to foster the use of waste streams of these industries and thereby reduce resource depletion and environmental impacts.

Opening of the Blood-Brain Barrier Using Low-Intensity Pulsed Ultrasound Enhances Responses to Immunotherapy in Preclinical Glioma Models.

The blood-brain barrier (BBB) inhibits adequate dosing/penetration of therapeutic agents to malignancies in the brain. Low-intensity pulsed ultrasound (LIPU) is a safe therapeutic method of temporary BBB disruption (BBBD) to enhance chemotherapeutic delivery to the tumor and surrounding brain parenchyma for treatment of glioblastoma. We investigated if LIPU could enhance therapeutic efficacy of anti-PD-1 in C57BL/6 mice bearing intracranial GL261 gliomas, epidermal growth factor receptor variant III (EGFRvIII) chimeric antigen receptor (CAR) T cells in NSG mice with EGFRvIII-U87 gliomas, and a genetically engineered antigen-presenting cell (APC)-based therapy producing the T-cell attracting chemokine CXCL10 in the GL261-bearing mice. Mice treated with anti-PD-1 and LIPU-induced BBBD had a median survival duration of 58 days compared with 39 days for mice treated with anti-PD-1, and long-term survivors all remained alive after contralateral hemisphere rechallenge. CAR T-cell administration with LIPU-induced BBBD resulted in significant increases in CAR T-cell delivery to the CNS after 24 (P < 0.005) and 72 (P < 0.001) hours and increased median survival by greater than 129%, in comparison with CAR T cells alone. Local deposition of CXCL10-secreting APCs in the glioma microenvironment with LIPU enhanced T-cell glioma infiltration during the therapeutic window (P = 0.004) and markedly enhanced survival (P < 0.05). LIPU increases immune therapeutic delivery to the tumor microenvironment with an associated increase in survival and is an emerging technique for enhancing novel therapies in the brain.

025 Sleep Disruption on an Orbital Shaker alters Glutamate in Prairie Vole Prefrontal Cortex

Abstract Introduction Glutamate concentrations in the cortex fluctuate with the sleep wake cycle in both rodents and humans. Altered glutamatergic signaling, as well as the early life onset of sleep disturbances have been implicated in neurodevelopmental disorders such as autism spectrum disorder. In order to study how sleep modulates glutamate activity in brain regions relevant to social behavior and development, we disrupted sleep in the socially monogamous prairie vole (Microtus ochrogaster) rodent species and quantified markers of glutamate neurotransmission within the prefrontal cortex, an area of the brain responsible for advanced cognition and complex social behaviors. Methods Male and female prairie voles were sleep disrupted using an orbital shaker to deliver automated gentle cage agitation at continuous intervals. Sleep was measured using EEG/EMG signals and paired with real time glutamate concentrations in the prefrontal cortex using an amperometric glutamate biosensor. This same method of sleep disruption was applied early in development (postnatal days 14–21) and the long term effects on brain development were quantified by examining glutamatergic synapses in adulthood. Results Consistent with previous research in rats, glutamate concentration in the prefrontal cortex increased during periods of wake in the prairie vole. Sleep disruption using the orbital shaker method resulted in brief cortical arousals and reduced time in REM sleep. When applied during development, early life sleep disruption resulted in long-term changes in both pre- and post-synaptic components of glutamatergic synapses in the prairie vole prefrontal cortex including increased density of immature spines. Conclusion In the prairie vole rodent model, sleep disruption on an orbital shaker produces a sleep, behavioral, and neurological phenotype that mirrors aspects of autism spectrum disorder including altered features of excitatory neurotransmission within the prefrontal cortex. Studies using this method of sleep disruption combined with real time biosensors for excitatory neurotransmitters will enhance our understanding of modifiable risk factors, such as sleep, that contribute to the altered development of glutamatergic synapses in the brain and their relationship to social behavior. Support (if any) NSF #1926818, VA CDA #IK2 BX002712, Portland VA Research Foundation, NIH NHLBI 5T32HL083808-10, VA Merit Review #I01BX001643

Polyvalent Bacterial Lysate Protects Against Pneumonia Independently of Neutrophils, IL-17A or Caspase-1 Activation.

Polyvalent bacterial lysates have been in use for decades for prevention and treatment of respiratory infections with reported clinical benefits. However, besides claims of broad immune activation, the mode of action is still a matter of debate. The lysates, formulated with the main bacterial species involved in respiratory infections, are commonly prepared by chemical or mechanical disruption of bacterial cells, what is believed influences the biological activity of the product. Here, we prepared two polyvalent lysates with the same composition but different method of bacterial cell disruption and evaluated their biological activity in a comparative fashion. We found that both bacterial lysates induce NF-kB activation in a MyD88 dependent manner, suggesting they work as TLR agonists. Further, we found that a single intranasal dose of any of the two lysates, is sufficient to protect against pneumococcal pneumonia, suggesting that they exert similar biological activity. We have previously shown that protection against pneumococcal pneumonia can also be induced by prior S. pneumoniae sub lethal infection or therapeutic treatment with a TLR5 agonist. Protection in those cases depends on neutrophil recruitment to the lungs, and can be associated with increased local expression of IL-17A. Here, we show that bacterial lysates exert protection against pneumococcal pneumonia independently of neutrophils, IL-17A or Caspase-1/11 activation, suggesting the existence of redundant mechanisms of protection. Trypsin-treated lysates afford protection to the same extent, suggesting that just small peptides suffice to exert the protective effect or that the molecules responsible for the protective effect are not proteins. Understanding the mechanism of action of bacterial lysates and deciphering the active components shall allow redesigning them with more precisely defined formulations and expanding their range of action.

Sub-surface metal loss defect detection using cold thermography and dynamic reference reconstruction (DRR)

Corrosion is considered a destructive phenomenon that affects almost all metals. Active infrared thermography is an online (no result delay) and non-intrusive (no process disruption) method of non-destructive testing (NDT), which has shown profound capabilities of detecting sub-surface metal loss. However, thermal reflections, non-uniform stimulation and lateral heat diffusion will remain as the most undesirable phenomena preventing the effective observation of sub-surface defects. This becomes more challenging when there is no a priori knowledge of the anomalies to effectively distinguish between defective and non-defective areas. In this work, cooling stimulation is considered as the thermal excitation mean as 1- a very few reports in this regard have been mentioned in the body of literature and 2- a dynamic setup was achieved that is found to be effective to minimise the possibility of disrupting reflections or artefacts registered by thermal camera similar to the case of using heating stimulation. A state-of-the-art prototype mechanism was manufactured. This equipment includes a carrier carrying a thermal camera and a cooling medium reservoir operating in reciprocating motion setup. This equipment is able to scan the test piece while cold stimulation is in operation, and immediately after that the camera registers the thermal evolution. An automated contrast enhancement pipeline using a variation of adaptive histogram equalisation (AHE) combined with principal component analysis (PCA) method was developed. The enhanced image results demonstrated the capability of accurately detecting sub-surface metal loss as low as 37.5% as well as an efficiently reconstructed reference (non-defective) area.

CRISPR-Cas9 cytidine and adenosine base editing of splice-sites mediates highly-efficient disruption of proteins in primary and immortalized cells

CRISPR-Cas9 cytidine and adenosine base editors (CBEs and ABEs) can disrupt genes without introducing double-stranded breaks by inactivating splice sites (BE-splice) or by introducing premature stop (pmSTOP) codons. However, no in-depth comparison of these methods or a modular tool for designing BE-splice sgRNAs exists. To address these needs, we develop SpliceR (http://z.umn.edu/spliceR) to design and rank BE-splice sgRNAs for any Ensembl annotated genome, and compared disruption approaches in T cells using a screen against the TCR-CD3 MHC Class I immune synapse. Among the targeted genes, we find that targeting splice-donors is the most reliable disruption method, followed by targeting splice-acceptors, and introducing pmSTOPs. Further, the CBE BE4 is more effective for disruption than the ABE ABE7.10, however this disparity is eliminated by employing ABE8e. Collectively, we demonstrate a robust method for gene disruption, accompanied by a modular design tool that is of use to basic and translational researchers alike.

Status and treatment of knee extension device disruption after total knee arthroplasty

To summarize the principle, classification, and treatment methods of knee extension device disruption after total knee arthroplasty (TKA). By extensively consulted the relevant domestic and abroad literature, the principle, classification, and treatment methods of injury according to different parts of the knee extension device after TKA were summarized and analyzed. The knee extension device disruption after TKA mainly occurs in the quadriceps tendon, patella tendon, and patella. Once the knee extension device is injured, it will seriously affect the functional recovery of the patient after surgery, resulting in delayed knee extension, limited range of motion, difficulty walking, and joint pain, etc. The current treatment methods are diverse, including conservative treatment and surgical treatment. Surgical treatment includes direct suture repair (traditional perosseous fixation and suture anchoring techniques), reconstruction and reinforcement repair (reconstruction using synthetic patches, autografts, and allografts). For the treatment of different parts, it is necessary to comprehensively consider the patient's knee joint tissue condition, the presence or absence of underlying diseases, and the presence or absence of donors. There is no uniform conclusion on the treatment of knee extension device disruption after TKA. Different injury situations need to be considered comprehensively to choose the appropriate treatment method.

PHYSICAL PRETREATMENT OF ULVA FASCIATA FOR ENHANCINGBIODIESEL PRODUCTION AND QUALITY

The green algae Ulva faciata was subjected to different physical pretreatments comprising thermal and mechanical techniques at different experimental conditions to state the most appropriate method of cell disruption for increasing the quantity of the extracted lipid and hence improve the quality of the produced biodiesel with low cost. Thermal pretreatment was autoclaving of either wet or dry algal biomass, while mechanical pretreatments include microwave and ultrasonication at different time intervals. The control was the alga without pretreatment extracted at optimum conditions: 60 min, 55oC, shaking speed at 250 rpm, &lt; 0.16 mm particle size with 25:1 v/w solvent to solid ratio. The results showed that the quantity of extracted lipids in case of using all physical pretreatments increased the Total fatty acids yield significantly by about 2-folds of the control for wet algae in hydrothermal treatment with optimum time of treatment 40 minutes, and 1.4 folds for dry algae in thermal pretreatment of the dried alga for 60minutes autoclaving period. The sharp increase by 2.2 folds of extracted lipids was recorded by microwave pretreatment for radiation period (5 min), while ultrasonication showed 2.1-fold increase in lipid yield at 15minutes ultrasound exposure time. Concerning the physical properties of the produced biodiesel after all physical pretreatments, the results indicated that the produced biodiesel had very high quality as all its properties are almost complied with the ASTM D6751 and EN14214 standards. These results were confirmed statistically where all physical pretreatments had high significant effect on fatty acids yield and Biodiesel properties.

Sputum processing by mechanical dissociation: A rapid alternative to traditional sputum assessment approaches.

Sputum cytology is currently the gold standard to evaluate cellular inflammation in the airways and phenotyping patients with airways diseases. Sputum eosinophil proportions have been used to guide treatment for moderate to severe asthma. Furthermore, raised sputum neutrophils are associated with poor disease control and impaired lung function in both asthma and COPD and small airways disease in cystic fibrosis. However, induced-sputum analysis is subjective and resource heavy, requiring dedicated specialist processing and assessment; this limits its utility in most clinical settings. Indirect blood eosinophil measures have been adopted in clinical care. However, there are currently no good peripheral blood biomarkers of airway neutrophils. A resource-light sputum processing approach could thus help integrate induced sputum more readily into routine clinical care. New mechanical disruption (MD) methods can rapidly obtain viable single cell suspensions from sputum samples. The aim of this study was to compare MD sputum processing to traditional methods for cell viability, granulocyte proportions and sputum cytokine analysis. Sputum plugs were split and processed using traditional methods and the MD method, and samples were then compared. The MD method produced a homogeneous cell suspension in 62s; 70min faster than the standard method used. No significant difference was seen between the cell viability (p=0.09), or the concentration of eosinophils (p=0.83), neutrophils (p=0.99) or interleukin-8 (p=0.86) using MD. This cost-effective method of sputum processing could provide a more pragmatic, sustainable means of directly monitoring the airway milieu. Therefore, we recommend this method be taken forward for further investigation.

CRISPR/Cas12a-mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus.

SummaryPurple non‐sulfur photosynthetic bacteria (PNSB) such as Rhodobacter capsulatus serve as a versatile platform for fundamental studies and various biotechnological applications. In this study, we sought to develop the class II RNA‐guided CRISPR/Cas12a system from Francisella novicida for genome editing and transcriptional regulation in R. capsulatus. Template‐free disruption method mediated by CRISPR/Cas12a reached ˜ 90% editing efficiency when targeting ccoO or nifH gene. When both genes were simultaneously edited, the multiplex editing efficiency reached > 63%. In addition, CRISPR interference (CRISPRi) using deactivated Cas12a was also evaluated using reporter genes egfp and lacZ, and the transcriptional repression efficiency reached ˜ 80%. In summary, our work represents the first report to develop CRISPR/Cas12a‐mediated genome editing and transcriptional regulation in R. capsulatus, which would greatly accelerate PNSB‐related researches.

Mechanical Cell Disruption Technologies for the Extraction of Dyes and Pigments from Microorganisms: A Review

The production of pigments using single cell microorganisms is gaining traction as a sustainable alternative to conventional syntheses, which rely, in no negligible proportions, on petrochemicals. In addition to depending on petroleum, these syntheses involved the use of toxic organic solvents, which may be inadequately disposed of across a range of industries, thus compounding the deleterious effects of fossil fuel exploitation. Literature suggests that notable research efforts in the area of sustainable pigment production using single cell microorganisms are focused on the production of pigments coveted for their interesting qualities, which transcend their mere capacity to dye various fabrics both natural and synthetic. As interest in sustainable pigment biosynthesis grows, the need to devise effective and efficient cell disruption processes becomes more pressing given that the viability of pigment biosynthesis is not only dependent on microorganisms’ yield in terms of production, but also on researchers’ ability to recover them. This review chiefly reports findings as to mechanical cell disruption methods, used individually or in various combinations, and their aptitude to recover biosynthetic pigments.

Current Pretreatment/Cell Disruption and Extraction Methods Used to Improve Intracellular Lipid Recovery from Oleaginous Yeasts.

The production of lipids from oleaginous yeasts involves several stages starting from cultivation and lipid accumulation, biomass harvesting and finally lipids extraction. However, the complex and relatively resistant cell wall of yeasts limits the full recovery of intracellular lipids and usually solvent extraction is not sufficient to effectively extract the lipid bodies. A pretreatment or cell disruption method is hence a prerequisite prior to solvent extraction. In general, there are no recovery methods that are equally efficient for different species of oleaginous yeasts. Each method adopts different mechanisms to disrupt cells and extract the lipids, thus a systematic evaluation is essential before choosing a particular method. In this review, mechanical (bead mill, ultrasonication, homogenization and microwave) and nonmechanical (enzyme, acid, base digestions and osmotic shock) methods that are currently used for the disruption or permeabilization of oleaginous yeasts are discussed based on their principle, application and feasibility, including their effects on the lipid yield. The attempts of using conventional and “green” solvents to selectively extract lipids are compared. Other emerging methods such as automated pressurized liquid extraction, supercritical fluid extraction and simultaneous in situ lipid recovery using capturing agents are also reviewed to facilitate the choice of more effective lipid recovery methods.

Microwave-Assisted One-Pot Lipid Extraction and Glycolipid Production from Oleaginous Yeast Saitozyma podzolica in Sugar Alcohol-Based Media.

Glycolipids are non-ionic surfactants occurring in numerous products of daily life. Due to their surface-activity, emulsifying properties, and foaming abilities, they can be applied in food, cosmetics, and pharmaceuticals. Enzymatic synthesis of glycolipids based on carbohydrates and free fatty acids or esters is often catalyzed using certain acyltransferases in reaction media of low water activity, e.g., organic solvents or notably Deep Eutectic Systems (DESs). Existing reports describing integrated processes for glycolipid production from renewables use many reaction steps, therefore this study aims at simplifying the procedure. By using microwave dielectric heating, DESs preparation was first accelerated considerably. A comparative study revealed a preparation time on average 16-fold faster than the conventional heating method in an incubator. Furthermore, lipids from robust oleaginous yeast biomass were successfully extracted up to 70% without using the pre-treatment method for cell disruption, limiting logically the energy input necessary for such process. Acidified DESs consisting of either xylitol or sorbitol and choline chloride mediated the one-pot process, allowing subsequent conversion of the lipids into mono-acylated palmitate, oleate, linoleate, and stearate sugar alcohol esters. Thus, we show strong evidence that addition of immobilized Candida antarctica Lipase B (Novozym 435®), in acidified DES mixture, enables a simplified and fast glycolipid synthesis using directly oleaginous yeast biomass.

Transportome-wide engineering of Saccharomyces cerevisiae

Synthetic biology enables the production of small molecules by recombinant microbes for pharma, food, and materials applications. The secretion of products reduces the cost of separation and purification, but it is challenging to engineer due to the limited understanding of the transporter proteins' functions. Here we describe a method for genome-wide transporter disruption that, in combination with a metabolite biosensor, enables the identification of transporters impacting the production of a given target metabolite in yeast Saccharomyces cerevisiae. We applied the method to study the transport of xenobiotic compounds, cis,cis-muconic acid (CCM), protocatechuic acid (PCA), and betaxanthins. We found 22 transporters that influenced the production of CCM or PCA. The transporter of the 12-spanner drug:H(+) antiporter (DHA1) family Tpo2p was further confirmed to import CCM and PCA in Xenopus expression assays. We also identified three transporter proteins (Qdr1p, Qdr2p, and Apl1p) involved in betaxanthins transport. In summary, the described method enables high-throughput transporter identification for small molecules in cell factories.

A real-time sleep scoring framework for closed-loop sleep manipulation in mice.

Subtle changes in sleep architecture can accompany and be symptomatic of many diseases or disorders. In order to probe and understand the complex interactions between sleep and health, the ability to model, track, and modulate sleep in preclinical animal models is vital. While various methods have been described for scoring experimental sleep recordings, few are designed to work in real time - a prerequisite for closed-loop sleep manipulation. In the present study, we have developed algorithms and software to classify sleep in real time and validated it on C57BL/6 mice (n=8). Hidden Markov models of baseline sleep dynamics were fitted using an unsupervised algorithm to electroencephalogram (EEG) and electromyogram (EMG) data for each mouse, and were able to classify sleep in a manner highly concordant with manual scoring (Cohen's Kappa >75%) up to 3weeks after model construction. This approach produced reasonably accurate estimates of common sleep metrics (proportion, mean duration, and number of bouts). After construction, the models were used to track sleep in real time and accomplish selective rapid eye movement (REM) sleep restriction by triggering non-invasive somatosensory stimulation. During REM restriction trials, REM bout duration was significantly reduced, and the classifier continued to perform satisfactorily despite the disrupted sleep patterns. The software can easily be tailored for use with other commercial or customised methods of sleep disruption (e.g. stir bar, optogenetic stimulation, etc.) and could serve as a robust platform to facilitate closed-loop experimentation. The source code and documentation are freely available upon request from the authors.

Mechanical/Physical Methods of Cell Disruption and Tissue Homogenization.

This chapter covers the various methods of mechanical cell disruption and tissue homogenization that are currently commercially available for processing small samples s<1mL) to larger multikilogram production quantities. These mechanical methods of lysing do not introduce chemicals or enzymes to the system. However, the energies required when using these "harsh," high mechanical energy methods can be enough to damage the very components being sought.The destruction of cell membranes and walls is effected by subjecting the cells (a) to shearing by liquid flow, (b) to exploding by pressure differences between inside and outside of cell, (c) to collision forces by impact of beads or paddles, or (d) a combination of these forces.Practical suggestions to optimize each method, where to acquire such equipment, and links to reference sources are included. Several novel technologies are presented.

Preparation of Cell Extracts for Purification of Proteins Expressed in Pichia pastoris.

Recovery of intracellular proteins requires disruption of the host cell before the target protein is extracted and isolated. Disruption methods vary depending on the type of cells, the total volume, and the number of samples being processed. For cells enveloped in cell walls (such as yeast), mild techniques such as hypotonic shock are not sufficient to achieve adequate lysis. More vigorous methods are often required. Although the preferred medium- or large-scale method of breaking yeast cells is mechanical shearing, lysis with the aid of glass beads in a BeadBeater is described here.

Candida albicans Bgl2p, Ecm33p, and Als1p proteins are involved in adhesion to saliva-coated hydroxyapatite

ABSTRACT Introduction: Candida albicans is an opportunistic pathogen that causes oral candidiasis. A previous study showed that Bgl2p and Ecm33p may mediate the interaction between the yeast and saliva-coated hydroxyapatite (SHA; a model for the tooth surface). This study investigated the roles of these cell wall proteins in the adherence of C. albicans to SHA beads. Methods: C. albicans BGL2 and ECM33 null mutants were generated from wild-type strain SC5314 by using the SAT1-flipper gene disruption method. A novel method based on labelling the yeast with Nile red, was used to investigate the adherence. Results: Adhesion of bgl2Δ and ecm33Δ null mutants to SHA beads was 76.4% and 64.8% of the wild-type strain, respectively. Interestingly, the adhesion of the bgl2Δ, ecm33Δ double mutant (87.7%) was higher than that of both single mutants. qRT-PCR analysis indicated that the ALS1 gene was over-expressed in the bgl2Δ, ecm33Δ strain. The triple null mutant showed a significantly reduced adherence to the beads, (37.6%), compared to the wild-type strain. Conclusion: Bgl2p and Ecm33p contributed to the interaction between C. albicans and SHA beads. Deletion of these genes triggered overexpression of the ALS1 gene in the bgl2Δ/ecm33Δ mutant strain, and deletion of all three genes caused a significant decrease in adhesion.

Disruption-Based Multiobjective Equilibrium Optimization Algorithm.

Nature-inspired computing has attracted huge attention since its origin, especially in the field of multiobjective optimization. This paper proposes a disruption-based multiobjective equilibrium optimization algorithm (DMOEOA). A novel mutation operator named layered disruption method is integrated into the proposed algorithm with the aim of enhancing the exploration and exploitation abilities of DMOEOA. To demonstrate the advantages of the proposed algorithm, various benchmarks have been selected with five different multiobjective optimization algorithms. The test results indicate that DMOEOA does exhibit better performances in these problems with a better balance between convergence and distribution. In addition, the new proposed algorithm is applied to the structural optimization of an elastic truss with the other five existing multiobjective optimization algorithms. The obtained results demonstrate that DMOEOA is not only an algorithm with good performance for benchmark problems but is also expected to have a wide application in real-world engineering optimization problems.