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3D-printing: an emerging and a revolutionary technology in pharmaceuticals

Singhvi G, Patil S, Girdhar V, Chellappan DK, Gupta G, Dua K

Panminerva Med, 2018 Dec;60(4):170-173.
PMID: 29856179 DOI: 10.23736/S0031-0808.18.03467-5

One of the novel and progressive technology employed in pharmaceutical manufacturing, design of medical device and tissue engineering is three-dimensional (3D) printing. 3D printing technologies provide great advantages in 3D scaffolds fabrication over traditional methods in the control of pore size, porosity, and interconnectivity. Various techniques of 3D-printing include powder bed fusion, fused deposition modeling, binder deposition, inkjet printing, photopolymerization and many others which are still evolving. 3D-printing technique been employed in developing immediate release products, various systems to deliver multiple release modalities etc. 3D printing has opened the door for new generation of customized drug delivery with built-in flexibility for safer and effective therapy. Our mini-review provides a quick snapshot on an overview of 3D printing, various techniques employed, applications and its advancements in pharmaceutical sciences.

Matched MeSH terms: Tissue Engineering/methods*
Fulltext A Review of Cell Adhesion Studies for Biomedical and Biological Applications

Khalili AA, Ahmad MR

Int J Mol Sci, 2015 Aug 05;16(8):18149-84.
PMID: 26251901 DOI: 10.3390/ijms160818149

Cell adhesion is essential in cell communication and regulation, and is of fundamental importance in the development and maintenance of tissues. The mechanical interactions between a cell and its extracellular matrix (ECM) can influence and control cell behavior and function. The essential function of cell adhesion has created tremendous interests in developing methods for measuring and studying cell adhesion properties. The study of cell adhesion could be categorized into cell adhesion attachment and detachment events. The study of cell adhesion has been widely explored via both events for many important purposes in cellular biology, biomedical, and engineering fields. Cell adhesion attachment and detachment events could be further grouped into the cell population and single cell approach. Various techniques to measure cell adhesion have been applied to many fields of study in order to gain understanding of cell signaling pathways, biomaterial studies for implantable sensors, artificial bone and tooth replacement, the development of tissue-on-a-chip and organ-on-a-chip in tissue engineering, the effects of biochemical treatments and environmental stimuli to the cell adhesion, the potential of drug treatments, cancer metastasis study, and the determination of the adhesion properties of normal and cancerous cells. This review discussed the overview of the available methods to study cell adhesion through attachment and detachment events.

Matched MeSH terms: Tissue Engineering/methods*
A Self-Replicating Linear DNA

Liew PS, Tan TH, Wong YC, Sim EUH, Lee CW, Narayanan K

ACS Synth Biol, 2020 04 17;9(4):804-813.
PMID: 32196315 DOI: 10.1021/acssynbio.9b00478

TelN and tos are a unique DNA linearization unit isolated from bacteriophage N15. While being transferable, the TelN cleaving-rejoining activities remained stable to function on tos in both bacterial and mammalian environments. However, TelN contribution in linear plasmid replication in mammalian cells remains unknown. Herein, we investigated the association of TelN in linear tos-containing DNA (tos-DNA) replication in mammalian cells. Additionally, the mammalian origin of replication (ori) that is well-known to initiate the replication event of plasmid vectors was also studied. In doing so, we identified that both TelN and mammalian initiation sites were essential for the replication of linear tos-DNA, determined by using methylation sensitive DpnI/MboI digestion and polymerase chain reaction (PCR) amplification approaches. Furthermore, we engineered the linear tos-DNA to be able to retain in mammalian cells using S/MAR technology. The resulting S/MAR containing tos-DNA was robust for at least 15 days, with (1) continuous tos-DNA replication, (2) correct splicing of gene transcripts, and (3) stable exogenous gene expression that was statistically comparable to the endogenous gene expression level. Understanding the activities of TelN and tos in mammalian cells can potentially provide insights for adapting this simple DNA linearization unit in developing novel genetic engineering tools, especially to the eukaryotic telomere/telomerase study.

Matched MeSH terms: Genetic Engineering/methods*
Fulltext A Systematic Analysis of Additive Manufacturing Techniques in the Bioengineering of In Vitro Cardiovascular Models

Mohanadas HP, Nair V, Doctor AA, Faudzi AAM, Tucker N, Ismail AF, et al.

Ann Biomed Eng, 2023 Nov;51(11):2365-2383.
PMID: 37466879 DOI: 10.1007/s10439-023-03322-x

Additive Manufacturing is noted for ease of product customization and short production run cost-effectiveness. As our global population approaches 8 billion, additive manufacturing has a future in maintaining and improving average human life expectancy for the same reasons that it has advantaged general manufacturing. In recent years, additive manufacturing has been applied to tissue engineering, regenerative medicine, and drug delivery. Additive Manufacturing combined with tissue engineering and biocompatibility studies offers future opportunities for various complex cardiovascular implants and surgeries. This paper is a comprehensive overview of current technological advancements in additive manufacturing with potential for cardiovascular application. The current limitations and prospects of the technology for cardiovascular applications are explored and evaluated.

Matched MeSH terms: Tissue Engineering/methods
Fulltext A comparative study on in vitro osteogenic priming potential of electron spun scaffold PLLA/HA/Col, PLLA/HA, and PLLA/Col for tissue engineering application

Balaji Raghavendran HR, Puvaneswary S, Talebian S, Murali MR, Raman Murali M, Naveen SV, et al.

PLoS One, 2014;9(8):e104389.
PMID: 25140798 DOI: 10.1371/journal.pone.0104389

A comparative study on the in vitro osteogenic potential of electrospun poly-L-lactide/hydroxyapatite/collagen (PLLA/HA/Col, PLLA/HA, and PLLA/Col) scaffolds was conducted. The morphology, chemical composition, and surface roughness of the fibrous scaffolds were examined. Furthermore, cell attachment, distribution, morphology, mineralization, extracellular matrix protein localization, and gene expression of human mesenchymal stromal cells (hMSCs) differentiated on the fibrous scaffolds PLLA/Col/HA, PLLA/Col, and PLLA/HA were also analyzed. The electrospun scaffolds with a diameter of 200-950 nm demonstrated well-formed interconnected fibrous network structure, which supported the growth of hMSCs. When compared with PLLA/H%A and PLLA/Col scaffolds, PLLA/Col/HA scaffolds presented a higher density of viable cells and significant upregulation of genes associated with osteogenic lineage, which were achieved without the use of specific medium or growth factors. These results were supported by the elevated levels of calcium, osteocalcin, and mineralization (P<0.05) observed at different time points (0, 7, 14, and 21 days). Furthermore, electron microscopic observations and fibronectin localization revealed that PLLA/Col/HA scaffolds exhibited superior osteoinductivity, when compared with PLLA/Col or PLLA/HA scaffolds. These findings indicated that the fibrous structure and synergistic action of Col and nano-HA with high-molecular-weight PLLA played a vital role in inducing osteogenic differentiation of hMSCs. The data obtained in this study demonstrated that the developed fibrous PLLA/Col/HA biocomposite scaffold may be supportive for stem cell based therapies for bone repair, when compared with the other two scaffolds.

Matched MeSH terms: Tissue Engineering/methods*
A review of evolution of electrospun tissue engineering scaffold: From two dimensions to three dimensions

Ngadiman NHA, Noordin MY, Idris A, Kurniawan D

Proc Inst Mech Eng H, 2017 Jul;231(7):597-616.
PMID: 28347262 DOI: 10.1177/0954411917699021

The potential of electrospinning process to fabricate ultrafine fibers as building blocks for tissue engineering scaffolds is well recognized. The scaffold construct produced by electrospinning process depends on the quality of the fibers. In electrospinning, material selection and parameter setting are among many factors that contribute to the quality of the ultrafine fibers, which eventually determine the performance of the tissue engineering scaffolds. The major challenge of conventional electrospun scaffolds is the nature of electrospinning process which can only produce two-dimensional electrospun mats, hence limiting their applications. Researchers have started to focus on overcoming this limitation by combining electrospinning with other techniques to fabricate three-dimensional scaffold constructs. This article reviews various polymeric materials and their composites/blends that have been successfully electrospun for tissue engineering scaffolds, their mechanical properties, and the various parameters settings that influence the fiber morphology. This review also highlights the secondary processes to electrospinning that have been used to develop three-dimensional tissue engineering scaffolds as well as the steps undertaken to overcome electrospinning limitations.

Matched MeSH terms: Tissue Engineering/methods*
A review of: application of synthetic scaffold in tissue engineering heart valves

Fallahiarezoudar E, Ahmadipourroudposht M, Idris A, Mohd Yusof N

Mater Sci Eng C Mater Biol Appl, 2015 Mar;48:556-65.
PMID: 25579957 DOI: 10.1016/j.msec.2014.12.016

The four heart valves represented in the mammalian hearts are responsible for maintaining unidirectional, non-hinder blood flow. The heart valve leaflets synchronically open and close approximately 4 million times a year and more than 3 billion times during the life. Valvular heart dysfunction is a significant cause of morbidity and mortality around the world. When one of the valves malfunctions, the medical choice is may be to replace the original valves with an artificial one. Currently, the mechanical and biological artificial valves are clinically used with some drawbacks. Tissue engineering heart valve concept represents a new technique to enhance the current model. In tissue engineering method, a three-dimensional scaffold is fabricated as the template for neo-tissue development. Appropriate cells are seeded to the matrix in vitro. Various approaches have been investigated either in scaffold biomaterials and fabrication techniques or cell source and cultivation methods. The available results of ongoing experiments indicate a promising future in this area (particularly in combination of bone marrow stem cells with synthetic scaffold), which can eliminate the need for lifelong anti-coagulation medication, durability and reoperation problems.

Matched MeSH terms: Tissue Engineering/methods*
Fulltext A review on Lactococcus lactis: from food to factory

Song AA, In LLA, Lim SHE, Rahim RA

Microb Cell Fact, 2017 04 04;16(1):55.
PMID: 28376880 DOI: 10.1186/s12934-017-0669-x

Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features of this gram-positive lactic acid non-colonizing gut bacteria include its generally recognized as safe (GRAS) status, probiotic properties, the absence of inclusion bodies and endotoxins, surface display and extracellular secretion technology, and a diverse selection of cloning and inducible expression vectors. This have made L. lactis a desirable and promising host on par with other well established model bacterial or yeast systems such as Escherichia coli, Saccharomyces [corrected] cerevisiae and Bacillus subtilis. In this article, we review recent technological advancements, challenges, future prospects and current diversified examples on the use of L. lactis as a microbial cell factory. Additionally, we will also highlight latest medical-based applications involving whole-cell L. lactis as a live delivery vector for the administration of therapeutics against both communicable and non-communicable diseases.

Matched MeSH terms: Genetic Engineering/methods*; Metabolic Engineering/methods*
Fulltext A study of computer-related upper limb discomfort and computer vision syndrome

Sen A, Richardson S

J Hum Ergol (Tokyo), 2007 Dec;36(2):45-50.
PMID: 18572794 DOI: 10.11183/jhe1972.36.2_45

Personal computers are one of the commonest office tools in Malaysia today. Their usage, even for three hours per day, leads to a health risk of developing Occupational Overuse Syndrome (OOS), Computer Vision Syndrome (CVS), low back pain, tension headaches and psychosocial stress. The study was conducted to investigate how a multiethnic society in Malaysia is coping with these problems that are increasing at a phenomenal rate in the west. This study investigated computer usage, awareness of ergonomic modifications of computer furniture and peripherals, symptoms of CVS and risk of developing OOS. A cross-sectional questionnaire study of 136 computer users was conducted on a sample population of university students and office staff. A 'Modified Rapid Upper Limb Assessment (RULA) for office work' technique was used for evaluation of OOS. The prevalence of CVS was surveyed incorporating a 10-point scoring system for each of its various symptoms. It was found that many were using standard keyboard and mouse without any ergonomic modifications. Around 50% of those with some low back pain did not have an adjustable backrest. Many users had higher RULA scores of the wrist and neck suggesting increased risk of developing OOS, which needed further intervention. Many (64%) were using refractive corrections and still had high scores of CVS commonly including eye fatigue, headache and burning sensation. The increase of CVS scores (suggesting more subjective symptoms) correlated with increase in computer usage spells. It was concluded that further onsite studies are needed, to follow up this survey to decrease the risks of developing CVS and OOS amongst young computer users.

Matched MeSH terms: Human Engineering/methods
A study of ecological sanitation as an integrated urban water supply system: case study of sustainable strategy for Kuching City, Sarawak, Malaysia

Seng DM, Putuhena FJ, Said S, Ling LP

J Water Health, 2009 Mar;7(1):169-84.
PMID: 18957785 DOI: 10.2166/wh.2009.103

A city consumes a large amount of water. Urban planning and development are becoming more compelling due to the fact of growing competition for water, which has lead to an increasing and conflicting demand. As such, investments in water supply, sanitation and water resources management is a strong potential for a solid return. A pilot project of greywater ecological treatment has been established in Kuching city since 2003. Such a treatment facility opens up an opportunity of wastewater reclamation for reuse as secondary sources of water for non-consumptive purposes. This paper aims to explore the potential of the intended purposes in the newly developed ecological treatment project. By utilizing the Wallingford Software model, InfoWorks WS (Water Supply) is employed to carry out a hydraulic modeling of a hypothetical greywater recycling system as an integrated part of the Kuching urban water supply, where the greywater is treated, recycled and reused in the domestic environment. The modeling efforts have shown water savings of about 40% from the investigated system reinstating that the system presents an alternative water source worth exploring in an urban environment.

Matched MeSH terms: Sanitary Engineering/methods*
A sustainable system of systems approach: a new HFE paradigm

Thatcher A, Yeow PH

Ergonomics, 2016;59(2):167-78.
PMID: 26307437 DOI: 10.1080/00140139.2015.1066876

Sustainability issues such as natural resource depletion, pollution and poor working conditions have no geographical boundaries in our interconnected world. To address these issues requires a paradigm shift within human factors and ergonomics (HFE), to think beyond a bounded, linear model understanding towards a broader systems framework. For this reason, we introduce a sustainable system of systems model that integrates the current hierarchical conceptualisation of possible interventions (i.e., micro-, meso- and macro-ergonomics) with important concepts from the sustainability literature, including the triple bottom line approach and the notion of time frames. Two practical examples from the HFE literature are presented to illustrate the model. The implications of this paradigm shift for HFE researchers and practitioners are discussed and include the long-term sustainability of the HFE community and comprehensive solutions to problems that consider the emergent issues that arise from this interconnected world.

Matched MeSH terms: Human Engineering/methods*
A transient assay to evaluate the expression of polyhydroxybutyrate genes regulated by oil palm mesocarp-specific promoter

Omidvar V, Siti Nor Akmar A, Marziah M, Maheran AA

Plant Cell Rep, 2008 Sep;27(9):1451-9.
PMID: 18563415 DOI: 10.1007/s00299-008-0565-2

The promoter of the oil palm metallothionein-like gene (MT3-A) demonstrated mesocarp-specific activity in functional analysis using transient expression assay of reporter gene in bombarded oil palm tissue slices. In order to investigate the tissue-specific expression of polyhydroxybutyrate (PHB) biosynthetic pathway genes, a multi-gene construct carrying PHB genes fused to the oil palm MT3-A promoter was co-transferred with a construct carrying GFP reporter gene using microprojectile bombardment targeting the mesocarp and leaf tissues of the oil palm. Transcriptional analysis using RT-PCR revealed successful transcription of all the three phbA, phbB, and phbC genes in transiently transformed mesocarp but not in transiently transformed leaf tissues. Furthermore, all the three expected sizes of PHB-encoded protein products were only detected in transiently transformed mesocarp tissues on a silver stained polyacrylamide gel. Western blot analysis using polyclonal antibody specific for phbB product confirmed successful translation of phbB mRNA transcript into protein product. This study provided valuable information, supporting the future engineering of PHB-producing transgenic palms.

Matched MeSH terms: Genetic Engineering/methods*
An endeavour from porous to 3D constructs bioceramics

Idris B, Rusnah M, Reusmaazran YM, Rohaida CH

Med J Malaysia, 2004 May;59 Suppl B:67-8.
PMID: 15468822
Matched MeSH terms: Tissue Engineering/methods*
Fulltext Antimicrobial quaternary ammonium organosilane cross-linked nanofibrous collagen scaffolds for tissue engineering

Dhand C, Balakrishnan Y, Ong ST, Dwivedi N, Venugopal JR, Harini S, et al.

Int J Nanomedicine, 2018;13:4473-4492.
PMID: 30122921 DOI: 10.2147/IJN.S159770

Introduction: In search for cross-linkers with multifunctional characteristics, the present work investigated the utility of quaternary ammonium organosilane (QOS) as a potential cross-linker for electrospun collagen nanofibers. We hypothesized that the quaternary ammonium ions improve the electrospinnability by reducing the surface tension and confer antimicrobial properties, while the formation of siloxane after alkaline hydrolysis could cross-link collagen and stimulate cell proliferation.
Materials and methods: QOS collagen nanofibers were electrospun by incorporating various concentrations of QOS (0.1%-10% w/w) and were cross-linked in situ after exposure to ammonium carbonate. The QOS cross-linked scaffolds were characterized and their biological properties were evaluated in terms of their biocompatibility, cellular adhesion and metabolic activity for primary human dermal fibroblasts and human fetal osteoblasts.
Results and discussion: The study revealed that 1) QOS cross-linking increased the flexibility of otherwise rigid collagen nanofibers and improved the thermal stability; 2) QOS cross-linked mats displayed potent antibacterial activity and 3) the biocompatibility of the composite mats depended on the amount of QOS present in dope solution - at low QOS concentrations (0.1% w/w), the mats promoted mammalian cell proliferation and growth, whereas at higher QOS concentrations, cytotoxic effect was observed.
Conclusion: This study demonstrates that QOS cross-linked mats possess anti-infective properties and confer niches for cellular growth and proliferation, thus offering a useful approach, which is important for hard and soft tissue engineering and regenerative medicine.

Matched MeSH terms: Tissue Engineering/methods*
Application of response surface methodology in optimization of electrospinning process to fabricate (ferrofluid/polyvinyl alcohol) magnetic nanofibers

Ahmadipourroudposht M, Fallahiarezoudar E, Yusof NM, Idris A

Mater Sci Eng C Mater Biol Appl, 2015 May;50:234-41.
PMID: 25746266 DOI: 10.1016/j.msec.2015.02.008

Magnetic nanofibers are composed of good dispersion of magnetic nanoparticles along an organic material. Magnetic nanofibers are potentially useful for composite reinforcement, bio-medical and tissue engineering. Nanofibers with the thinner diameter have to result in higher rigidity and tensile strength due to better alignments of lamellae along the fiber axis. In this study, the performance of electrospinning process was explained using response surface methodology (RSM) during fabrication of magnetic nanofibers using polyvinyl alcohol (PVA) as a shelter for (γ-Fe2O3) nanoparticles where the parameters investigated were flow rate, applied voltage, distance between needle and collector and collector rotating speed. The response variable was diameter distribution. The two parameters flow rate and applied voltage in primary evaluation were distinguished as significant factors. Central composite design was applied to optimize the variable of diameter distribution. Quadratic estimated model developed for diameter distribution indicated the optimum conditions to be flow rate of 0.25 ml/h at voltage of 45 kV while the distance and rotating speed are at 8 cm and 1500 rps respectively. The obtained model was verified successfully by the confirmation experiments.

Matched MeSH terms: Tissue Engineering/methods*
Fulltext Application of stem cells in tissue engineering for defense medicine

Ude CC, Miskon A, Idrus RBH, Abu Bakar MB

Mil Med Res, 2018 02 26;5(1):7.
PMID: 29502528 DOI: 10.1186/s40779-018-0154-9

The dynamic nature of modern warfare, including threats and injuries faced by soldiers, necessitates the development of countermeasures that address a wide variety of injuries. Tissue engineering has emerged as a field with the potential to provide contemporary solutions. In this review, discussions focus on the applications of stem cells in tissue engineering to address health risks frequently faced by combatants at war. Human development depends intimately on stem cells, the mysterious precursor to every kind of cell in the body that, with proper instruction, can grow and differentiate into any new tissue or organ. Recent reports have suggested the greater therapeutic effects of the anti-inflammatory, trophic, paracrine and immune-modulatory functions associated with these cells, which induce them to restore normal healing and tissue regeneration by modulating immune reactions, regulating inflammation, and suppressing fibrosis. Therefore, the use of stem cells holds significant promise for the treatment of many battlefield injuries and their complications. These applications include the treatment of injuries to the skin, sensory organs, nervous system tissues, the musculoskeletal system, circulatory/pulmonary tissues and genitals/testicles and of acute radiation syndrome and the development of novel biosensors. The new research developments in these areas suggest that solutions are being developed to reduce critical consequences of wounds and exposures suffered in warfare. Current military applications of stem cell-based therapies are already saving the lives of soldiers who would have died in previous conflicts. Injuries that would have resulted in deaths previously now result in wounds today; similarly, today's permanent wounds may be reduced to tomorrow's bad memories with further advances in stem cell-based therapies.

Matched MeSH terms: Tissue Engineering/methods
Autologous chondrocyte transplantation in the repair of full-thickness focal cartilage damage in rabbits

Kamarul T, Selvaratnam L, Masjuddin T, Ab-Rahim S, Ng C, Chan KY, et al.

J Orthop Surg (Hong Kong), 2008 Aug;16(2):230-6.
PMID: 18725678

To compare the efficacy of autologous chondrocyte transplantation (ACT) versus non-operative measures for cartilage repair in rabbits.

Matched MeSH terms: Tissue Engineering/methods*
Autologous implantation of bilayered tissue-engineered respiratory epithelium for tracheal mucosal regenesis in a sheep model

Mohd Heikal MY, Aminuddin BS, Jeevanan J, Chen HC, Sharifah SH, Ruszymah BH

Cells Tissues Organs (Print), 2010;192(5):292-302.
PMID: 20616535 DOI: 10.1159/000318675

The objective of this study was to regenerate the tracheal epithelium using autologous nasal respiratory epithelial cells in a sheep model. Respiratory epithelium and fibroblast cells were harvested from nasal turbinates and cultured for 1 week. After confluence, respiratory epithelium and fibroblast cells were suspended in autologous fibrin polymerized separately to form a tissue-engineered respiratory epithelial construct (TEREC). A 3 × 2 cm² tracheal mucosal defect was created, and implanted with TEREC and titanium mesh as a temporary scaffold. The control groups were divided into 2 groups: polymerized autologous fibrin devoid of cells (group 1), and no construct implanted (group 2). All sheep were euthanized at 4 weeks of implantation. Gross observation of the trachea showed minimal luminal stenosis formation in the experimental group compared to the control groups. Macroscopic evaluation revealed significant mucosal fibrosis in control group 1 (71.8%) as compared to the experimental group (7%). Hematoxylin and eosin staining revealed the presence of minimal overgrowth of fibrous connective tissue covered by respiratory epithelium. A positive red fluorescence staining of PKH26 on engineered tissue 4 weeks after implantation confirmed the presence of cultured nasal respiratory epithelial cells intercalated with native tracheal epithelial cells. Scanning electron microscopy showed the presence of short microvilli representing immature cilia on the surface of the epithelium. Our study showed that TEREC was a good replacement for a tracheal mucosal defect and was able to promote natural regenesis of the tracheal epithelium with minimal fibrosis. This study highlighted a new technique in the treatment of tracheal stenosis.

Matched MeSH terms: Tissue Engineering/methods*
Autologous versus pooled human serum for articular chondrocyte growth

Munirah S, Ruszymah BH, Samsudin OC, Badrul AH, Azmi B, Aminuddin BS

J Orthop Surg (Hong Kong), 2008 Aug;16(2):220-9.
PMID: 18725677

To evaluate the effect of autologous human serum (AHS) versus pooled human serum (PHS) versus foetal bovine serum (FBS) for growth of articular chondrocytes and formation of chondrocytefibrin constructs.

Matched MeSH terms: Tissue Engineering/methods*
Fulltext Basic fibroblast growth factor with human serum supplementation: enhancement of human chondrocyte proliferation and promotion of cartilage regeneration

Chua KH, Aminuddin BS, Fuzina NH, Ruszymah BH

Singapore Med J, 2007 Apr;48(4):324-32.
PMID: 17384880

The objectives of this study were to determine the optimum concentration of basic fibroblast growth factor (bFGF) in foetal bovine serum (FBS) or human serum (HS) supplemented medium for adult human nasal septum chondrocyte culture and to evaluate the potential of cartilage regeneration.

Matched MeSH terms: Tissue Engineering/methods*

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