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Bioengineering, Volume 8, Issue 3 (March 2021) – 12 articles

Cover Story (view full-size image): CO2 removal via membrane oxygenators has become an important clinical technique. In order to further optimize oxygenators, accurate prediction of the CO2 removal rate is required. It can be determined either by measuring the CO2 content in the exhaust gas of the oxygenator (sweep flow-based) or by using blood gas analyzer data and a CO2 solubility model (blood-based). In this study, we determined the CO2 removal rate of an oxygenator by using both methods in in vitro experiments with bovine blood, as well as in in vivo experiments with porcine blood. This allows direct comparison and evaluation of the two methods. Furthermore, the suitability of four unique CO2 solubility models, with different levels of complexity, was investigated. View this paper
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Open AccessArticle
SCRaMbLE: A Study of Its Robustness and Challenges through Enhancement of Hygromycin B Resistance in a Semi-Synthetic Yeast
Bioengineering 2021, 8(3), 42; https://doi.org/10.3390/bioengineering8030042 - 23 Mar 2021
Viewed by 639
Abstract
Recent advances in synthetic genomics launched the ambitious goal of generating the first synthetic designer eukaryote, based on the model organism Saccharomyces cerevisiae (Sc2.0). Excitingly, the Sc2.0 project is now nearing its completion and SCRaMbLE, an accelerated evolution tool implemented by the integration [...] Read more.
Recent advances in synthetic genomics launched the ambitious goal of generating the first synthetic designer eukaryote, based on the model organism Saccharomyces cerevisiae (Sc2.0). Excitingly, the Sc2.0 project is now nearing its completion and SCRaMbLE, an accelerated evolution tool implemented by the integration of symmetrical loxP sites (loxPSym) downstream of almost every non-essential gene, is arguably the most applicable synthetic genome-wide alteration to date. The SCRaMbLE system offers the capability to perform rapid genome diversification, providing huge potential for targeted strain improvement. Here we describe how SCRaMbLE can evolve a semi-synthetic yeast strain housing the synthetic chromosome II (synII) to generate hygromycin B resistant genotypes. Exploiting long-read nanopore sequencing, we show that all structural variations are due to recombination between loxP sites, with no off-target effects. We also highlight a phenomenon imposed on SCRaMbLE termed “essential raft”, where a fragment flanked by a pair of loxPSym sites can move within the genome but cannot be removed due to essentiality restrictions. Despite this, SCRaMbLE was able to explore the genomic space and produce alternative structural compositions that resulted in an increased hygromycin B resistance in the synII strain. We show that among the rearrangements generated via SCRaMbLE, deletions of YBR219C and YBR220C contribute to hygromycin B resistance phenotypes. However, the hygromycin B resistance provided by SCRaMbLEd genomes showed significant improvement when compared to corresponding single deletions, demonstrating the importance of the complex structural variations generated by SCRaMbLE to improve hygromycin B resistance. We anticipate that SCRaMbLE and its successors will be an invaluable tool to predict and evaluate the emergence of antibiotic resistance in yeast. Full article
(This article belongs to the Special Issue From Yeast to Biotechnology)
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Open AccessArticle
Spectral Decomposition of the Flow and Characterization of the Sound Signals through Stenoses with Different Levels of Severity
Bioengineering 2021, 8(3), 41; https://doi.org/10.3390/bioengineering8030041 - 19 Mar 2021
Viewed by 357
Abstract
Treatments of atherosclerosis depend on the severity of the disease at the diagnosis time. Non-invasive diagnosis techniques, capable of detecting stenosis at early stages, are essential to reduce associated costs and mortality rates. We used computational fluid dynamics and acoustics analysis to extensively [...] Read more.
Treatments of atherosclerosis depend on the severity of the disease at the diagnosis time. Non-invasive diagnosis techniques, capable of detecting stenosis at early stages, are essential to reduce associated costs and mortality rates. We used computational fluid dynamics and acoustics analysis to extensively investigate the sound sources arising from high-turbulent fluctuating flow through stenosis. The frequency spectral analysis and proper orthogonal decomposition unveiled the frequency contents of the fluctuations for different severities and decomposed the flow into several frequency bandwidths. Results showed that high-intensity turbulent pressure fluctuations appeared inside the stenosis for severities above 70%, concentrated at plaque surface, and immediately in the post-stenotic region. Analysis of these fluctuations with the progression of the stenosis indicated that (a) there was a distinct break frequency for each severity level, ranging from 40 to 230 Hz, (b) acoustic spatial-frequency maps demonstrated the variation of the frequency content with respect to the distance from the stenosis, and (c) high-energy, high-frequency fluctuations existed inside the stenosis only for severe cases. This information can be essential for predicting the severity level of progressive stenosis, comprehending the nature of the sound sources, and determining the location of the stenosis with respect to the point of measurements. Full article
(This article belongs to the Special Issue Advances in Multivariate Physiological Signal Analysis)
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Open AccessArticle
P3CMQA: Single-Model Quality Assessment Using 3DCNN with Profile-Based Features
Bioengineering 2021, 8(3), 40; https://doi.org/10.3390/bioengineering8030040 - 19 Mar 2021
Viewed by 373
Abstract
Model quality assessment (MQA), which selects near-native structures from structure models, is an important process in protein tertiary structure prediction. The three-dimensional convolution neural network (3DCNN) was applied to the task, but the performance was comparable to existing methods because it used only [...] Read more.
Model quality assessment (MQA), which selects near-native structures from structure models, is an important process in protein tertiary structure prediction. The three-dimensional convolution neural network (3DCNN) was applied to the task, but the performance was comparable to existing methods because it used only atom-type features as the input. Thus, we added sequence profile-based features, which are also used in other methods, to improve the performance. We developed a single-model MQA method for protein structures based on 3DCNN using sequence profile-based features, namely, P3CMQA. Performance evaluation using a CASP13 dataset showed that profile-based features improved the assessment performance, and the proposed method was better than currently available single-model MQA methods, including the previous 3DCNN-based method. We also implemented a web-interface of the method to make it more user-friendly. Full article
(This article belongs to the Special Issue New Bioinformatics Tools)
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Open AccessReview
Collagen-Based Electrospun Materials for Tissue Engineering: A Systematic Review
Bioengineering 2021, 8(3), 39; https://doi.org/10.3390/bioengineering8030039 - 18 Mar 2021
Viewed by 361
Abstract
Collagen is a key component of the extracellular matrix (ECM) in organs and tissues throughout the body and is used for many tissue engineering applications. Electrospinning of collagen can produce scaffolds in a wide variety of shapes, fiber diameters and porosities to match [...] Read more.
Collagen is a key component of the extracellular matrix (ECM) in organs and tissues throughout the body and is used for many tissue engineering applications. Electrospinning of collagen can produce scaffolds in a wide variety of shapes, fiber diameters and porosities to match that of the native ECM. This systematic review aims to pool data from available manuscripts on electrospun collagen and tissue engineering to provide insight into the connection between source material, solvent, crosslinking method and functional outcomes. D-banding was most often observed in electrospun collagen formed using collagen type I isolated from calfskin, often isolated within the laboratory, with short solution solubilization times. All physical and chemical methods of crosslinking utilized imparted resistance to degradation and increased strength. Cytotoxicity was observed at high concentrations of crosslinking agents and when abbreviated rinsing protocols were utilized. Collagen and collagen-based scaffolds were capable of forming engineered tissues in vitro and in vivo with high similarity to the native structures. Full article
(This article belongs to the Special Issue Biomedical Applications of Collagen)
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Open AccessArticle
Surface Engineered Iron Oxide Nanoparticles Generated by Inert Gas Condensation for Biomedical Applications
Bioengineering 2021, 8(3), 38; https://doi.org/10.3390/bioengineering8030038 - 15 Mar 2021
Viewed by 384
Abstract
Despite the lifesaving medical discoveries of the last century, there is still an urgent need to improve the curative rate and reduce mortality in many fatal diseases such as cancer. One of the main requirements is to find new ways to deliver therapeutics/drugs [...] Read more.
Despite the lifesaving medical discoveries of the last century, there is still an urgent need to improve the curative rate and reduce mortality in many fatal diseases such as cancer. One of the main requirements is to find new ways to deliver therapeutics/drugs more efficiently and only to affected tissues/organs. An exciting new technology is nanomaterials which are being widely investigated as potential nanocarriers to achieve localized drug delivery that would improve therapy and reduce adverse drug side effects. Among all the nanocarriers, iron oxide nanoparticles (IONPs) are one of the most promising as, thanks to their paramagnetic/superparamagnetic properties, they can be easily modified with chemical and biological functions and can be visualized inside the body by magnetic resonance imaging (MRI), while delivering the targeted therapy. Therefore, iron oxide nanoparticles were produced here with a novel method and their properties for potential applications in both diagnostics and therapeutics were investigated. The novel method involves production of free standing IONPs by inert gas condensation via the Mantis NanoGen Trio physical vapor deposition system. The IONPs were first sputtered and deposited on plasma cleaned, polyethylene glycol (PEG) coated silicon wafers. Surface modification of the cleaned wafer with PEG enabled deposition of free-standing IONPs, as once produced, the soft-landed IONPs were suspended by dissolution of the PEG layer in water. Transmission electron microscopic (TEM) characterization revealed free standing, iron oxide nanoparticles with size < 20 nm within a polymer matrix. The nanoparticles were analyzed also by Atomic Force Microscope (AFM), Dynamic Light Scattering (DLS) and NanoSight Nanoparticle Tacking Analysis (NTA). Therefore, our work confirms that inert gas condensation by the Mantis NanoGen Trio physical vapor deposition sputtering at room temperature can be successfully used as a scalable, reproducible process to prepare free-standing IONPs. The PEG- IONPs produced in this work do not require further purification and thanks to their tunable narrow size distribution have potential to be a powerful tool for biomedical applications. Full article
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Open AccessReview
Hydrogel Models with Stiffness Gradients for Interrogating Pancreatic Cancer Cell Fate
Bioengineering 2021, 8(3), 37; https://doi.org/10.3390/bioengineering8030037 - 13 Mar 2021
Viewed by 429
Abstract
Pancreatic ductal adenocarcinoma (PDAC) is the most common type of pancreatic cancer and has seen only modest improvements in patient survival rate over the past few decades. PDAC is highly aggressive and resistant to chemotherapy, owing to the presence of a dense and [...] Read more.
Pancreatic ductal adenocarcinoma (PDAC) is the most common type of pancreatic cancer and has seen only modest improvements in patient survival rate over the past few decades. PDAC is highly aggressive and resistant to chemotherapy, owing to the presence of a dense and hypovascularized fibrotic tissue, which is composed of stromal cells and extracellular matrices. Increase deposition and crosslinking of matrices by stromal cells lead to a heterogeneous microenvironment that aids in PDAC development. In the past decade, various hydrogel-based, in vitro tumor models have been developed to mimic and recapitulate aspects of the tumor microenvironment in PDAC. Advances in hydrogel chemistry and engineering should provide a venue for discovering new insights regarding how matrix properties govern PDAC cell growth, migration, invasion, and drug resistance. These engineered hydrogels are ideal for understanding how variation in matrix properties contributes to the progressiveness of cancer cells, including durotaxis, the directional migration of cells in response to a stiffness gradient. This review surveys the various hydrogel-based, in vitro tumor models and the methods to generate gradient stiffness for studying migration and other cancer cell fate processes in PDAC. Full article
(This article belongs to the Special Issue Cell–Biomaterial Interactions)
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Open AccessArticle
Saccharomyces cerevisiae Concentrates Subtoxic Copper onto Cell Wall from Solid Media Containing Reducing Sugars as Carbon Source
Bioengineering 2021, 8(3), 36; https://doi.org/10.3390/bioengineering8030036 - 06 Mar 2021
Viewed by 562
Abstract
Copper is essential for life, but it can be deleterious in concentrations that surpass the physiological limits. Copper pollution is related to widespread human activities, such as viticulture and wine production. To unravel aspects of how organisms cope with copper insults, we used [...] Read more.
Copper is essential for life, but it can be deleterious in concentrations that surpass the physiological limits. Copper pollution is related to widespread human activities, such as viticulture and wine production. To unravel aspects of how organisms cope with copper insults, we used Saccharomyces cerevisiae as a model for adaptation to high but subtoxic concentrations of copper. We found that S. cerevisiae cells could tolerate high copper concentration by forming deposits on the cell wall and that the copper-containing deposits accumulated predominantly when cells were grown statically on media prepared with reducing sugars (glucose, galactose) as sole carbon source, but not on media containing nonreducing carbon sources, such as glycerol or lactate. Exposing cells to copper in liquid media under strong agitation prevented the formation of copper-containing deposits at the cell wall. Disruption of low-affinity copper intake through the plasma membrane increased the potential of the cell to form copper deposits on the cell surface. These results imply that biotechnology problems caused by high copper concentration can be tackled by selecting yeast strains and conditions to allow the removal of excess copper from various contaminated sites in the forms of solid deposits which do not penetrate the cell. Full article
(This article belongs to the Special Issue From Yeast to Biotechnology)
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Open AccessArticle
Deep Neural Networks and Transfer Learning on a Multivariate Physiological Signal Dataset
Bioengineering 2021, 8(3), 35; https://doi.org/10.3390/bioengineering8030035 - 06 Mar 2021
Viewed by 506
Abstract
While Deep Neural Networks (DNNs) and Transfer Learning (TL) have greatly contributed to several medical and clinical disciplines, the application to multivariate physiological datasets is still limited. Current examples mainly focus on one physiological signal and can only utilise applications that are customised [...] Read more.
While Deep Neural Networks (DNNs) and Transfer Learning (TL) have greatly contributed to several medical and clinical disciplines, the application to multivariate physiological datasets is still limited. Current examples mainly focus on one physiological signal and can only utilise applications that are customised for that specific measure, thus it limits the possibility of transferring the trained DNN to other domains. In this study, we composed a dataset (n=813) of six different types of physiological signals (Electrocardiogram, Electrodermal activity, Electromyogram, Photoplethysmogram, Respiration and Acceleration). Signals were collected from 232 subjects using four different acquisition devices. We used a DNN to classify the type of physiological signal and to demonstrate how the TL approach allows the exploitation of the efficiency of DNNs in other domains. After the DNN was trained to optimally classify the type of signal, the features that were automatically extracted by the DNN were used to classify the type of device used for the acquisition using a Support Vector Machine. The dataset, the code and the trained parameters of the DNN are made publicly available to encourage the adoption of DNN and TL in applications with multivariate physiological signals. Full article
(This article belongs to the Special Issue Advances in Multivariate Physiological Signal Analysis)
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Open AccessArticle
Spectral Decomposition and Sound Source Localization of Highly Disturbed Flow through a Severe Arterial Stenosis
Bioengineering 2021, 8(3), 34; https://doi.org/10.3390/bioengineering8030034 - 04 Mar 2021
Cited by 1 | Viewed by 580
Abstract
For the early detection of atherosclerosis, it is imperative to explore the capabilities of new, effective noninvasive diagnosis techniques to significantly reduce the associated treatment costs and mortality rates. In this study, a multifaceted comprehensive approach involving advanced computational fluid dynamics combined with [...] Read more.
For the early detection of atherosclerosis, it is imperative to explore the capabilities of new, effective noninvasive diagnosis techniques to significantly reduce the associated treatment costs and mortality rates. In this study, a multifaceted comprehensive approach involving advanced computational fluid dynamics combined with signal processing techniques was exploited to investigate the highly turbulent fluctuating flow through arterial stenosis. The focus was on localizing high-energy mechano-acoustic source potential to transmit to the epidermal surface. The flow analysis results showed the existence of turbulent pressure fluctuations inside the stenosis and in the post-stenotic region. After analyzing the turbulent kinetic energy and pressure fluctuations on the flow centerline and the vessel wall, the point of maximum excitation in the flow was observed around two diameters downstream of the stenosis within the fluctuating zone. It was also found that the concentration of pressure fluctuation closer to the wall was higher inside the stenosis compared to the post-stenotic region. Additionally, the visualization of the most energetic proper orthogonal decomposition (POD) mode and spectral decomposition of the flow indicated that the break frequencies ranged from 80 to 220 Hz and were correlated to the eddies generated within these regions. Full article
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Open AccessArticle
Suitable CO2 Solubility Models for Determination of the CO2 Removal Performance of Oxygenators
Bioengineering 2021, 8(3), 33; https://doi.org/10.3390/bioengineering8030033 - 02 Mar 2021
Viewed by 534
Abstract
CO2 removal via membrane oxygenators during lung protective ventilation has become a reliable clinical technique. For further optimization of oxygenators, accurate prediction of the CO2 removal rate is necessary. It can either be determined by measuring the CO2 content in [...] Read more.
CO2 removal via membrane oxygenators during lung protective ventilation has become a reliable clinical technique. For further optimization of oxygenators, accurate prediction of the CO2 removal rate is necessary. It can either be determined by measuring the CO2 content in the exhaust gas of the oxygenator (sweep flow-based) or using blood gas analyzer data and a CO2 solubility model (blood-based). In this study, we determined the CO2 removal rate of a prototype oxygenator utilizing both methods in in vitro trials with bovine and in vivo trials with porcine blood. While the sweep flow-based method is reliably accurate, the blood-based method depends on the accuracy of the solubility model. In this work, we quantified performances of four different solubility models by calculating the deviation of the CO2 removal rates determined by both methods. Obtained data suggest that the simplest model (Loeppky) performs better than the more complex ones (May, Siggaard-Anderson, and Zierenberg). The models of May, Siggaard-Anderson, and Zierenberg show a significantly better performance for in vitro bovine blood data than for in vivo porcine blood data. Furthermore, the suitability of the Loeppky model parameters for bovine blood (in vitro) and porcine blood (in vivo) is evaluated. Full article
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Open AccessArticle
Variation of Passive Biomechanical Properties of the Small Intestine along Its Length: Microstructure-Based Characterization
Bioengineering 2021, 8(3), 32; https://doi.org/10.3390/bioengineering8030032 - 26 Feb 2021
Viewed by 473
Abstract
Multiaxial testing of the small intestinal wall is critical for understanding its biomechanical properties and defining material models, but limited data and material models are available. The aim of the present study was to develop a microstructure-based material model for the small intestine [...] Read more.
Multiaxial testing of the small intestinal wall is critical for understanding its biomechanical properties and defining material models, but limited data and material models are available. The aim of the present study was to develop a microstructure-based material model for the small intestine and test whether there was a significant variation in the passive biomechanical properties along the length of the organ. Rat tissue was cut into eight segments that underwent inflation/extension testing, and their nonlinearly hyper-elastic and anisotropic response was characterized by a fiber-reinforced model. Extensive parametric analysis showed a non-significant contribution to the model of the isotropic matrix and circumferential-fiber family, leading also to severe over-parameterization. Such issues were not apparent with the reduced neo-Hookean and (axial and diagonal)-fiber family model, that provided equally accurate fitting results. Absence from the model of either the axial or diagonal-fiber families led to ill representations of the force- and pressure-diameter data, respectively. The primary direction of anisotropy, designated by the estimated orientation angle of diagonal-fiber families, was about 35° to the axial direction, corroborating prior microscopic observations of submucosal collagen-fiber orientation. The estimated model parameters varied across and within the duodenum, jejunum, and ileum, corroborating histologically assessed segmental differences in layer thicknesses. Full article
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Open AccessArticle
Microbial Interactions as Drivers of a Nitrification Process in a Chemostat
Bioengineering 2021, 8(3), 31; https://doi.org/10.3390/bioengineering8030031 - 25 Feb 2021
Viewed by 545
Abstract
This article deals with the inclusion of microbial ecology measurements such as abundances of operational taxonomic units in bioprocess modelling. The first part presents the mathematical analysis of a model that may be framed within the class of Lotka–Volterra models fitted to experimental [...] Read more.
This article deals with the inclusion of microbial ecology measurements such as abundances of operational taxonomic units in bioprocess modelling. The first part presents the mathematical analysis of a model that may be framed within the class of Lotka–Volterra models fitted to experimental data in a chemostat setting where a nitrification process was operated for over 500 days. The limitations and the insights of such an approach are discussed. In the second part, the use of an optimal tracking technique (developed within the framework of control theory) for the integration of data from genetic sequencing in chemostat models is presented. The optimal tracking revisits the data used in the aforementioned chemostat setting. The resulting model is an explanatory model, not a predictive one, it is able to reconstruct the different forms of nitrogen in the reactor by using the abundances of the operational taxonomic units, providing some insights into the growth rate of microbes in a complex community. Full article
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