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Leveraging in vivo animal models of tendon loading to inform tissue engineering approaches. 利用肌腱加载的活体动物模型为组织工程方法提供信息。
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-07 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1449372
Samantha Muscat, Anne E C Nichols

Tendon injuries disrupt successful transmission of force between muscle and bone, resulting in reduced mobility, increased pain, and significantly reduced quality of life for affected patients. There are currently no targeted treatments to improve tendon healing beyond conservative methods such as rest and physical therapy. Tissue engineering approaches hold great promise for designing instructive biomaterials that could improve tendon healing or for generating replacement graft tissue. More recently, engineered microphysiological systems to model tendon injuries have been used to identify therapeutic targets. Despite these advances, current tissue engineering efforts that aim to regenerate, replace, or model injured tendons have largely failed due in large part to a lack of understanding of how the mechanical environment of the tendon influences tissue homeostasis and how altered mechanical loading can promote or prevent disease progression. This review article draws inspiration from what is known about tendon loading from in vivo animal models and identifies key metrics that can be used to benchmark success in tissue engineering applications. Finally, we highlight important challenges and opportunities for the field of tendon tissue engineering that should be taken into consideration in designing engineered platforms to understand or improve tendon healing.

肌腱损伤会破坏肌肉和骨骼之间成功的力量传递,导致患者活动能力下降、疼痛加剧,并大大降低生活质量。除了休息和物理治疗等保守方法外,目前还没有针对性的治疗方法来改善肌腱愈合。组织工程方法在设计可改善肌腱愈合或生成替代移植组织的指导性生物材料方面大有可为。最近,用于模拟肌腱损伤的工程微生理系统已被用于确定治疗目标。尽管取得了这些进展,但目前旨在再生、替代或模拟损伤肌腱的组织工程工作大多以失败告终,这在很大程度上是由于人们对肌腱的机械环境如何影响组织稳态以及改变机械负荷如何促进或预防疾病进展缺乏了解。这篇综述文章从体内动物模型对肌腱加载的了解中汲取灵感,并确定了可用于衡量组织工程应用成功与否的关键指标。最后,我们强调了肌腱组织工程领域的重要挑战和机遇,在设计工程平台以了解或改善肌腱愈合时应考虑到这些挑战和机遇。
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引用次数: 0
Effective removal of host cell-derived nucleic acids bound to hepatitis B core antigen virus-like particles by heparin chromatography. 用肝素层析法有效去除与乙型肝炎核心抗原病毒样颗粒结合的宿主细胞衍生核酸。
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-03 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1475918
Angela Valentic, Jürgen Hubbuch

Virus-like particles (VLPs) show considerable potential for a wide array of therapeutic applications, spanning from vaccines targeting infectious diseases to applications in cancer immunotherapy and drug delivery. In the context of hepatitis B core antigen (HBcAg) VLPs, a promising candidate for gene delivery approaches, the naturally occurring nucleic acid (NA) binding region is commonly utilized for effective binding of various types of therapeutic nucleic acids (NAther). During formation of the HBcAg VLPs, host cell-derived nucleic acids (NAhc) might be associated to the NA binding region, and are thus encapsulated into the VLPs. Following a VLP harvest, the NAhc need to be removed effectively before loading the VLP with NAther. Various techniques reported in literature for this NAhc removal, including enzymatic treatments, alkaline treatment, and lithium chloride precipitation, lack quantitative evidence of sufficient NAhc removal accompanied by a subsequent high VLP protein recovery. In this study, we present a novel heparin chromatography-based process for effective NAhc removal from HBcAg VLPs. Six HBcAg VLP constructs with varying lengths of the NA binding region and diverse NAhc loadings were subjected to evaluation. Process performance was thoroughly examined through NAhc removal and VLP protein recovery analyses. Hereby, reversed phase chromatography combined with UV/Vis spectroscopy, as well as silica spin column-based chromatography coupled with dye-based fluorescence assay were employed. Additionally, alternative process variants, comprising sulfate chromatography and additional nuclease treatments, were investigated. Comparative analyses were conducted with LiCl precipitation and alkaline treatment procedures to ascertain the efficacy of the newly developed chromatography-based methods. Results revealed the superior performance of the heparin chromatography procedure in achieving high NAhc removal and concurrent VLP protein recovery. Furthermore, nuanced relationships between NA binding region length and NAhc removal efficiency were elucidated. Hereby, the construct Cp157 surpassed the other constructs in the heparin process by demonstrating high NAhc removal and VLP protein recovery. Among the other process variants minimal performance variations were observed for the selected constructs Cp157 and Cp183. However, the heparin chromatography-based process consistently outperformed other methods, underscoring its superiority in NAhc removal and VLP protein recovery.

病毒样颗粒(VLPs)在广泛的治疗应用中显示出相当大的潜力,从针对传染病的疫苗到癌症免疫疗法和给药应用,无所不包。乙型肝炎核心抗原(HBcAg)VLP 是一种很有希望的基因递送方法,在乙型肝炎核心抗原(HBcAg)VLP 中,通常利用天然存在的核酸(NA)结合区来有效结合各种类型的治疗核酸(NAther)。在 HBcAg VLPs 的形成过程中,宿主细胞衍生的核酸(NAhc)可能与 NA 结合区相关联,从而被包裹到 VLPs 中。收获 VLP 后,需要先有效去除 NAhc,然后再将 NAther 装入 VLP。文献中报道的各种去除NAhc的技术,包括酶处理、碱处理和氯化锂沉淀等,都缺乏足够去除NAhc的定量证据,同时也缺乏随后高VLP蛋白回收率的定量证据。在本研究中,我们提出了一种基于肝素层析的新型工艺,可有效去除 HBcAg VLP 中的 NAhc。六种 HBcAg VLP 构建物的 NA 结合区长度各不相同,NAhc 负载也各不相同,我们对它们进行了评估。通过NAhc去除率和VLP蛋白回收率分析,对工艺性能进行了全面检测。为此,采用了反相色谱法结合紫外/可见光谱法,以及硅胶旋柱色谱法结合染料荧光测定法。此外,还研究了硫酸层析和额外核酸酶处理的替代工艺变体。与氯化锂沉淀和碱性处理程序进行了比较分析,以确定新开发的色谱法的功效。结果显示,肝素色谱法在实现高 NAhc 去除率和同时 VLP 蛋白回收率方面表现出色。此外,还阐明了 NA 结合区长度与 NAhc 去除效率之间的微妙关系。因此,在肝素工艺中,构建体 Cp157 的 NAhc 去除率和 VLP 蛋白回收率均高于其他构建体。在其他工艺变体中,所选构建体 Cp157 和 Cp183 的性能差异极小。然而,基于肝素色谱法的工艺始终优于其他方法,凸显了其在去除 NAhc 和回收 VLP 蛋白方面的优势。
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引用次数: 0
Biodielectrics: old wine in a new bottle? 生物电力:新瓶装旧酒?
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-03 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1458668
Hema Dinesh Barnana, Syed A M Tofail, Krittish Roy, Charlie O'Mahony, Veronika Hidaši Turiničová, Maroš Gregor, Ehtsham Ul Haq

Biodielectrics is a subset of biological and/or bioinspired materials that has brought a huge transformation in the advancement of medical science, such as localized drug delivery in cancer therapeutics, health monitoring, bone and nerve repair, tissue engineering and use in other nanoelectromechanical systems (NEMS). While biodielectrics has long been used in the field of electrical insulation for over a century, polar dielectric properties of biological building blocks have not been well understood at the fundamental building block level. In this review article, we provide a brief overview of dielectric properties of biological building blocks and its hierarchical organisations to include polar dielectric properties such as piezo, pyro, and ferroelectricity. This review article also discusses recent trends, scope, and potential applications of these dielectrics in science and technology. We highlight electromechanical properties embedded in rationally designed organic assemblies, and the challenges and opportunities inherent in mapping from molecular amino acid building blocks to macroscopic analogs of biological fibers and tissues, in pursuit of sustainable materials for next-generation technologies.

生物电学是生物和/或生物启发材料的一个分支,它为医学科学的发展带来了巨大的变革,如癌症治疗中的局部给药、健康监测、骨骼和神经修复、组织工程以及在其他纳米机电系统(NEMS)中的应用。虽然生物电学在电绝缘领域的应用已有一个多世纪的历史,但人们对生物构件的极性介电性能在基本构件层面上的了解还不够深入。在这篇综述文章中,我们简要概述了生物构件的介电性能及其分层组织,包括压电、热电和铁电等极性介电性能。这篇综述文章还讨论了这些电介质在科学和技术领域的最新趋势、范围和潜在应用。我们重点介绍了合理设计的有机组合体所蕴含的机电特性,以及从氨基酸分子构件到生物纤维和组织的宏观类似物的映射过程中所蕴含的挑战和机遇,以寻求下一代技术所需的可持续材料。
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引用次数: 0
Study on the effect of phenoxyethanol-citric acid pretreatment for the enzymatic hydrolysis of bamboo residues. 研究苯氧乙醇-柠檬酸预处理对酶水解竹渣的影响。
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-03 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1483025
Yan Cheng, Xiaoxue Zhao, Ruolin Li, Jili Liao, Caoxing Huang

This study investigated the biphasic phenoxyethanol-citric acid (PECA) pretreatment for bamboo residues (BRs) and its corresponding effects on the enzymatic hydrolysis performance. It is found that increasing the concentration of citric acid in the pretreatment system from 2.5% to 15% greatly enhanced the delignification and xylan removal for BRs. Consequently, the enzymatic hydrolysis yield of pretreated BRs significantly enhanced, increasing from 12.4% to 58.2% and 28.0%72.4% when the concentration of citric acid was increased from 2.5% to 15.0% at 160°C and 170°C, respectively. The characterization results from cellulose crystallinity, accessibility, and hydrophobicity of pretreated bamboo residues indicated that their changes possessed a beneficial performance on the enzymatic hydrolysis yield, which could result from the synergistic removal of lignin and xylan. The Chrastil model analysis showed that pretreatment at higher conditions resulted in the pretreated BRs possessing weaker diffusion resistance for cellulase, which is attributed to its higher enzymatic hydrolysis yield.

本研究探讨了双相苯氧乙醇-柠檬酸(PECA)预处理竹渣(BRs)的方法及其对酶水解性能的影响。研究发现,将预处理系统中柠檬酸的浓度从 2.5% 提高到 15%,可大大提高竹渣的脱木素和木聚糖能力。因此,当柠檬酸的浓度从 2.5% 增加到 15.0%,温度分别为 160°C 和 170°C 时,预处理过的 BR 的酶水解产率明显提高,分别从 12.4% 增加到 58.2% 和 28.0%72.4%。预处理竹渣的纤维素结晶度、可及性和疏水性的表征结果表明,它们的变化对酶水解产率有好处,这可能是由于协同去除木质素和木聚糖的结果。Chrastil 模型分析表明,在较高条件下进行预处理,预处理后的竹残片对纤维素酶的扩散阻力较弱,因此酶水解产率较高。
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引用次数: 0
Improvement of the gait deviation index for spinal cord injury to broaden its applicability: the reduced gait deviation index for spinal cord injury (rSCI-GDI). 改进脊髓损伤步态偏离指数,扩大其适用范围:脊髓损伤步态偏离指数缩减版(rSCI-GDI)。
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-02 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1431596
Diana Herrera-Valenzuela, Isabel Sinovas-Alonso, Ana de Los Reyes, Ángel Gil-Agudo, Antonio J Del-Ama

Background: The SCI-GDI is an accurate and effective metric to summarize gait kinematics in adults with SCI. It is usually computed with the information registered with a photogrammetry system because it requires accurate information of pelvic and hip movement in the three anatomic planes, which is hard to record with simpler systems. Additionally, due to being developed from the GDI, the SCI-GDI is built upon nine joint movements selected for a pediatric population with cerebral palsy, for which the GDI was originally developed, but those nine movements are not necessarily as meaningful for adults with SCI. Nevertheless, pelvic movement and hip rotation have been proven to have low reliability even when acquired with gold-standard photogrammetry systems. Additionally, the use of photogrammetry is limited in real-life scenarios and when used with rehabilitation technologies, which limits the use of the SCI-GDI to evaluate gait in alternative scenarios to gait laboratories and to evaluate technologies for gait assistance. This research aimed to improve the SCI-GDI to broaden its applicability beyond the use of photogrammetry.

Methods: An exploration of the mathematical relevance of each joint movement included in the original GDI for the performance of the metric is performed. Considering the results obtained and the clinical relevance of each of the 9 joints used to compute the SCI-GDI in the gait pattern of the SCI population, a more adaptable SCI-GDI is proposed using four joint movements that can be precisely captured with simpler systems than photogrammetry: sagittal planes of hip, knee and ankle and hip abduction/adduction.

Results: The reduced SCI-GDI (rSCI-GDI) effectively represents gait variability of adults with SCI as does the SCI-GDI, while providing more generalizable results and equivalent or stronger correlations with clinical tests validated in the population. During the derivation of the improved index, it was demonstrated that pelvic movements, hip rotation, and foot progression angle introduce high variability to the dataset of gait patterns of the adult population with SCI, but they have low relevance to characterize gait kinematics of this population. The rSCI-GDI can be calculated using the 14-feature vectorial basis included in the electronic addendum provided.

背景:SCI-GDI 是总结患有 SCI 的成人步态运动学的准确而有效的指标。它通常使用摄影测量系统登记的信息进行计算,因为它需要骨盆和髋关节在三个解剖平面上运动的准确信息,而简单的系统很难记录这些信息。此外,由于 SCI-GDI 是在 GDI 的基础上发展而来的,因此它建立在九个关节运动的基础上,而这九个关节运动最初是为脑瘫患儿开发的,但这九个关节运动对于患有 SCI 的成年人来说并不一定有意义。尽管如此,骨盆运动和髋关节旋转已被证明可靠性较低,即使使用黄金标准摄影测量系统进行采集也是如此。此外,摄影测量法在现实生活场景和康复技术中的应用有限,这限制了 SCI-GDI 在步态实验室以外的其他场景中评估步态和评估步态辅助技术的应用。这项研究旨在改进 SCI-GDI,以扩大其在摄影测量之外的适用性:方法:对原始 GDI 中包含的每个关节运动与指标性能的数学相关性进行了探讨。考虑到所获得的结果以及用于计算 SCI-GDI 的 9 个关节在 SCI 患者步态模式中的临床相关性,提出了一个适应性更强的 SCI-GDI,使用四个关节运动,这四个关节运动可以用比摄影测量更简单的系统精确捕捉:髋、膝、踝的矢状面以及髋关节内收/外展:结果:简化的 SCI-GDI (rSCI-GDI)与 SCI-GDI 一样有效地反映了患有 SCI 的成人的步态变异性,同时提供了更具普遍性的结果,并与在人群中验证的临床测试具有同等或更强的相关性。在推导改进指数的过程中,我们发现骨盆运动、髋关节旋转和足前进角会给患有 SCI 的成人步态模式数据集带来很大的变异性,但它们与描述该人群步态运动学特征的相关性较低。rSCI-GDI可以使用所提供电子附录中的14特征矢量基础进行计算。
{"title":"Improvement of the gait deviation index for spinal cord injury to broaden its applicability: the reduced gait deviation index for spinal cord injury (rSCI-GDI).","authors":"Diana Herrera-Valenzuela, Isabel Sinovas-Alonso, Ana de Los Reyes, Ángel Gil-Agudo, Antonio J Del-Ama","doi":"10.3389/fbioe.2024.1431596","DOIUrl":"https://doi.org/10.3389/fbioe.2024.1431596","url":null,"abstract":"<p><strong>Background: </strong>The SCI-GDI is an accurate and effective metric to summarize gait kinematics in adults with SCI. It is usually computed with the information registered with a photogrammetry system because it requires accurate information of pelvic and hip movement in the three anatomic planes, which is hard to record with simpler systems. Additionally, due to being developed from the GDI, the SCI-GDI is built upon nine joint movements selected for a pediatric population with cerebral palsy, for which the GDI was originally developed, but those nine movements are not necessarily as meaningful for adults with SCI. Nevertheless, pelvic movement and hip rotation have been proven to have low reliability even when acquired with gold-standard photogrammetry systems. Additionally, the use of photogrammetry is limited in real-life scenarios and when used with rehabilitation technologies, which limits the use of the SCI-GDI to evaluate gait in alternative scenarios to gait laboratories and to evaluate technologies for gait assistance. This research aimed to improve the SCI-GDI to broaden its applicability beyond the use of photogrammetry.</p><p><strong>Methods: </strong>An exploration of the mathematical relevance of each joint movement included in the original GDI for the performance of the metric is performed. Considering the results obtained and the clinical relevance of each of the 9 joints used to compute the SCI-GDI in the gait pattern of the SCI population, a more adaptable SCI-GDI is proposed using four joint movements that can be precisely captured with simpler systems than photogrammetry: sagittal planes of hip, knee and ankle and hip abduction/adduction.</p><p><strong>Results: </strong>The reduced SCI-GDI (rSCI-GDI) effectively represents gait variability of adults with SCI as does the SCI-GDI, while providing more generalizable results and equivalent or stronger correlations with clinical tests validated in the population. During the derivation of the improved index, it was demonstrated that pelvic movements, hip rotation, and foot progression angle introduce high variability to the dataset of gait patterns of the adult population with SCI, but they have low relevance to characterize gait kinematics of this population. The rSCI-GDI can be calculated using the 14-feature vectorial basis included in the electronic addendum provided.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11480198/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142461840","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Continuous Process Verification 4.0 application in upstream: adaptiveness implementation managed by AI in the hypoxic bioprocess of the Pichia pastoris cell factory. 持续过程验证 4.0 在上游的应用:在 Pichia pastoris 细胞工厂的缺氧生物过程中通过人工智能管理适应性的实施。
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-02 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1439638
Arnau Gasset, Joeri Van Wijngaarden, Ferran Mirabent, Albert Sales-Vallverdú, Xavier Garcia-Ortega, José Luis Montesinos-Seguí, Toni Manzano, Francisco Valero

The experimental approach developed in this research demonstrated how the cloud, the Internet of Things (IoT), edge computing, and Artificial Intelligence (AI), considered key technologies in Industry 4.0, provide the expected horizon for adaptive vision in Continued Process Verification (CPV), the final stage of Process Validation (PV). Pichia pastoris producing Candida rugosa lipase 1 under the regulation of the constitutive GAP promoter was selected as an experimental bioprocess. The bioprocess worked under hypoxic conditions in carbon-limited fed-batch cultures through a physiological control based on the respiratory quotient (RQ). In this novel bioprocess, a digital twin (DT) was built and successfully tested. The implementation of online sensors worked as a bridge between the microorganism and AI models, to provide predictions from the edge and the cloud. AI models emulated the metabolism of Pichia based on critical process parameters and actionable factors to achieve the expected quality attributes. This innovative AI-aided Adaptive-Proportional Control strategy (AI-APC) improved the reproducibility comparing to a Manual-Heuristic Control strategy (MHC), showing better performance than the Boolean-Logic-Controller (BLC) tested. The accuracy, indicated by the Mean Relative Error (MRE), was for the AI-APC lower than 4%, better than the obtained for MHC (10%) and BLC (5%). Moreover, in terms of precision, the same trend was observed when comparing the Root Mean Square Deviation (RMSD) values, becoming lower as the complexity of the controller increases. The successful automatic real time control of the bioprocess orchestrated by AI models proved the 4.0 capabilities brought by the adaptive concept and its validity in biopharmaceutical upstream operations.

本研究开发的实验方法展示了被视为工业 4.0 关键技术的云、物联网(IoT)、边缘计算和人工智能(AI)如何为持续过程验证(CPV)(过程验证(PV)的最后阶段)中的自适应视觉提供预期的前景。在组成型 GAP 启动子的调控下,生产白色念珠菌脂肪酶 1 的 Pichia pastoris 被选为实验性生物工艺。通过基于呼吸商数(RQ)的生理控制,该生物工艺在缺氧条件下的限碳喂料批量培养中运行。在这一新型生物过程中,建立并成功测试了数字孪生(DT)。在线传感器的实施在微生物和人工智能模型之间架起了一座桥梁,提供来自边缘和云端的预测。人工智能模型根据关键工艺参数和可操作因素模拟 Pichia 的新陈代谢,以实现预期的质量属性。这种创新的人工智能辅助自适应比例控制策略(AI-APC)与人工神经网络控制策略(MHC)相比,提高了可重复性,显示出比所测试的布尔逻辑控制器(BLC)更好的性能。以平均相对误差(MRE)表示,AI-APC 的精确度低于 4%,优于 MHC(10%)和 BLC(5%)。此外,在精确度方面,通过比较均方根偏差(RMSD)值也可以观察到相同的趋势,即随着控制器复杂度的增加,精确度也会降低。人工智能模型成功地对生物过程进行自动实时控制,证明了自适应概念带来的 4.0 功能及其在生物制药上游操作中的有效性。
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引用次数: 0
Potential of a novel brine-struvite-based growth medium for sustainable biomass and phycocyanin production by Arthrospira platensis. 基于盐水-吸附剂的新型生长培养基对板蓝根节肢动物可持续生产生物量和藻蓝蛋白的潜力。
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-02 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1466978
Stephan S W Ende, Albert S Beyer, Reham Ebaid, Mostafa Elshobary, Mafalda C Almeida, Cynthia Couto, Kit W Chew, Tamara Schwenkler, Joachim Henjes

Nutrient recovery is crucial for sustainability as it helps to recycle valuable resources, reduce environmental pollution, and promote the efficient use of natural materials in various agricultural and industrial processes. The present study investigated the impact of using brine and struvite as sustainable nutrient sources on the growth and c-phycocyanin (C-PC) production by the cyanobacterium Arthrospira platensis. Three modified growth media were compared to the standard SAG-spirul medium under yellow-white light [YLT], and blue-white light [BLT]. In the modified medium BSI, a struvite solution was utilized to replace dipotassium phosphate, while diluted brine was used to replace NaCl and de-ionized H2O. For BSII, struvite and brine were used as in BSI, with elimination of the micronutrient from the solution. In BSIII, no other nutrient sources than bicarbonate-buffer were used in addition to struvite and brine. For each medium, A. platensis was cultivated and incubated under YLT or BLT till the stationary phase. The results showed that the combinations of brine and struvite did not have any significant negative impact on the growth rates in BSIII. However, adding struvite as a phosphorus source boosted C-PC production just as effectively as YLT, with boosting biomass yield, unlike when only BLT was used. In conclusion, the brine/struvite-based media resulted in high biomass productivity with higher C-PC yields, making it an ideal growth medium for commercial sustainable C-PC production.

养分回收对可持续发展至关重要,因为它有助于循环利用宝贵的资源,减少环境污染,并促进各种农业和工业生产过程中天然材料的有效利用。本研究调查了使用盐水和硬石膏作为可持续营养源对蓝藻节杆菌的生长和 c-phycocyanin (C-PC) 生产的影响。在黄白光[YLT]和蓝白光[BLT]下,将三种改良生长培养基与标准 SAG-螺旋培养基进行了比较。在改良培养基 BSI 中,用硬石膏溶液代替磷酸氢二钾,用稀盐水代替氯化钠和去离子水。在 BSII 培养基中,与 BSI 培养基一样,也使用了硬石膏和盐水,但取消了溶液中的微量营养元素。在 BSIII 中,除了重碳酸盐缓冲液和盐水外,没有使用其他营养源。在每种培养基中,A. platensis 都在 YLT 或 BLT 条件下培养至静止期。结果表明,盐水和硬石膏的组合对 BSIII 的生长率没有明显的负面影响。然而,与只使用 BLT 时不同的是,添加硬石膏作为磷源与 YLT 一样有效地促进了 C-PC 的生产,并提高了生物量产量。总之,以盐水/硬石膏为基础的培养基具有较高的生物量生产率和较高的 C-PC 产量,是商业化可持续 C-PC 生产的理想生长培养基。
{"title":"Potential of a novel brine-struvite-based growth medium for sustainable biomass and phycocyanin production by <i>Arthrospira platensis</i>.","authors":"Stephan S W Ende, Albert S Beyer, Reham Ebaid, Mostafa Elshobary, Mafalda C Almeida, Cynthia Couto, Kit W Chew, Tamara Schwenkler, Joachim Henjes","doi":"10.3389/fbioe.2024.1466978","DOIUrl":"https://doi.org/10.3389/fbioe.2024.1466978","url":null,"abstract":"<p><p>Nutrient recovery is crucial for sustainability as it helps to recycle valuable resources, reduce environmental pollution, and promote the efficient use of natural materials in various agricultural and industrial processes. The present study investigated the impact of using brine and struvite as sustainable nutrient sources on the growth and c-phycocyanin (C-PC) production by the cyanobacterium <i>Arthrospira platensis</i>. Three modified growth media were compared to the standard SAG-spirul medium under yellow-white light [YLT], and blue-white light [BLT]. In the modified medium BSI, a struvite solution was utilized to replace dipotassium phosphate, while diluted brine was used to replace NaCl and de-ionized H<sub>2</sub>O. For BSII, struvite and brine were used as in BSI, with elimination of the micronutrient from the solution. In BSIII, no other nutrient sources than bicarbonate-buffer were used in addition to struvite and brine. For each medium, <i>A. platensis</i> was cultivated and incubated under YLT or BLT till the stationary phase. The results showed that the combinations of brine and struvite did not have any significant negative impact on the growth rates in BSIII. However, adding struvite as a phosphorus source boosted C-PC production just as effectively as YLT, with boosting biomass yield, unlike when only BLT was used. In conclusion, the brine/struvite-based media resulted in high biomass productivity with higher C-PC yields, making it an ideal growth medium for commercial sustainable C-PC production.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.3,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11479874/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142461855","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Metabolic engineering of hairy root cultures in Beta vulgaris for enhanced production of vanillin, 4-hydroxybenzoic acid, and vanillyl alcohol. 为提高香兰素、4-羟基苯甲酸和香草醇的产量而对红豆杉毛根培养物进行代谢工程改造。
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-02 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1435190
Zakir Husain, Zafar Iqbal Warsi, Sana Khan, Ganesan Mahendran, Shama Afroz, Ashish Chandran, Praveen Kumar Kashyap, Kahkashan Khatoon, Gazala Parween, Sudeep Tandon, Laiq Ur Rahman

The flavor of vanilla is a complex blend of compounds, with vanillin as the most prominent, along with vanillyl alcohol and 4-hydroxybenzoic acid. Natural vanillin extracted from vanilla beans is expensive, so researchers use heterologous synthesis to produce nature-identical vanillin in plant hosts. Consequently, alternative traditional farming and gathering methods are required to bridge the significant disparity between supply and demand. The current research successfully developed a method to induce hairy root formation from leaves. It integrated the Vanillin synthase (VpVAN) gene into transgenic hairy root lines of Beta vulgaris, synthesizing vanillin-related compounds. The presence of the VpVAN gene in transgenic roots was confirmed using PCR analysis. Additionally, RT-qPCR analysis demonstrated the expression of the VpVAN gene in the transgenic root lines. The transgenic hairy root clones H1, H2, and H5 showed enhanced vanillin production, vanillyl alcohol, and 4-hydroxybenzoic acid. Elicitation with methyl jasmonate (MJ) and salicylic acid (SA) further improved the production of these compounds in B. vulgaris hairy roots. The maximum hairy root biomass was observed after 60 days, with the maximum synthesis of vanillin and 4-hydroxybenzoic acid obtained from hairy root clones H5 and HR2, respectively. Vanillyl alcohol HR2 was obtained on the 45th day of cultivation. Elicitation with wound-associated hormone methyl jasmonate and salicylic acid enhanced the yield of vanillin, vanillyl alcohol, and 4-hydroxybenzoic acid, with a 215-fold increase in vanillin, a 13-fold increase in vanillyl alcohol, and a 21 fold increase in 4-hydroxybenzoic acid. The study results indicate that establishing transgenic hairy root cultures with the VpVAN gene is a promising alternative method for enhancing the production of vanilla flavor compounds such as vanillin, vanillyl alcohol, and 4-hydroxybenzoic acid. A cost-effective protocol has been developed to mass-produce phenolic compounds using a hairy root culture of B. vulgaris. This approach addresses the increasing demand for these substances while reducing the cost of natural vanillin production, making it suitable for industrial-scale applications.

香草的味道由多种复杂的化合物混合而成,其中最突出的是香兰素,此外还有香草醇和 4-羟基苯甲酸。从香草豆中提取的天然香兰素价格昂贵,因此研究人员采用异源合成法,在植物宿主体内生产与天然相同的香兰素。因此,需要采用其他传统种植和采集方法来弥补供需之间的巨大差距。目前的研究成功开发了一种从叶片诱导毛根形成的方法。该研究将香兰素合成酶(VpVAN)基因整合到转基因毛细根系中,合成香兰素相关化合物。通过 PCR 分析证实了转基因根中存在 VpVAN 基因。此外,RT-qPCR 分析也证明了 VpVAN 基因在转基因根系中的表达。转基因毛根克隆 H1、H2 和 H5 的香兰素、香草醇和 4-羟基苯甲酸产量均有所提高。茉莉酸甲酯(MJ)和水杨酸(SA)的激发进一步提高了粗壮毛根中这些化合物的产量。60 天后观察到毛状根生物量最大,毛状根克隆 H5 和 HR2 分别获得了最大的香兰素和 4-羟基苯甲酸合成量。香草醇 HR2 是在培养的第 45 天获得的。伤口相关激素茉莉酸甲酯和水杨酸的激发提高了香兰素、香草醇和 4-羟基苯甲酸的产量,其中香兰素增加了 215 倍,香草醇增加了 13 倍,4-羟基苯甲酸增加了 21 倍。研究结果表明,用 VpVAN 基因建立转基因毛根培养物是提高香草香味化合物(如香兰素、香草醇和 4-羟基苯甲酸)产量的一种很有前途的替代方法。目前已开发出一种具有成本效益的方案,利用毛根培养法大规模生产酚类化合物。这种方法既能满足对这些物质日益增长的需求,又能降低天然香兰素的生产成本,适合工业规模的应用。
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引用次数: 0
Establishment of a platform based on dual RPA combined with CRISPR/Cas12a for the detection of Klebsiella pneumoniae and its KPC resistance gene. 建立一个基于双 RPA 与 CRISPR/Cas12a 结合的平台,用于检测肺炎克雷伯氏菌及其 KPC 耐药基因。
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-02 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1447963
Meiying Tan, Xueli Yi, Chuan Liao, Zihan Zhou, Baoyan Ren, Lina Liang, Xuebin Li, Guijiang Wei

Carbapenem resistant Klebsiella pneumoniae (CRKP) can cause serious hospital- and community-acquired infections. Treatment for CRKP infection is limited, resulting in prolonged hospitalization and high consultation costs. The KPC genotype has the highest detection rate of CRKP, and its mortality rate is higher than the overall mortality rate of CRKP. However, traditional testing methods have disadvantages such as long time and reliance on complex and sophisticated instruments, which are not conducive to rapid screening for CRKP. Therefore, this study aimed to establish a detection platform for early screening of CRKP so that effective antimicrobial therapy could be administered promptly to prevent the widespread spread of CRKP. We integrated dual RPA with CRISPR/Cas12a to establish a dual platform for the detection of K. pneumoniae (Kp) rcsA-specific gene and KPC resistance gene. Four result reading methods were established, including fluorescence detection (FD), blue light irradiation detection (BLID), ultraviolet irradiation detection (UID), and lateral flow test strips (LFTS). For the rcsA gene, the LOD of FD was 1 × 10 pg/μL, and the other three methods could detect 1 × 101 pg/μL of bacterial DNA. As for the KPC gene, four resultant readout methods were able to detect 1 × 102 pg/μL of bacterial DNA. In 59 clinical strains tested, the dual RPA-CRISPR/Cas12a detection of the rcsA had 100% sensitivity, specificity, and accuracy compared to the culture method. Compared with the drug sensitivity test, the sensitivity of dual RPA-CRISPR/Cas12a detection for the KPC was 85.71%, the specificity was 100%, and the accuracy was 94.92%. In summary, our dual RPA-CRISPR/Cas12a platform proved to be rapid, precise, and convenient for the efficient detection of Kp with KPC in the laboratory or at the point of care.

耐碳酸培南肺炎克雷伯菌(CRKP)可导致严重的医院和社区获得性感染。CRKP 感染的治疗方法有限,导致住院时间延长和高昂的诊疗费用。KPC 基因型在 CRKP 中的检出率最高,其死亡率也高于 CRKP 的总体死亡率。然而,传统检测方法存在时间长、依赖复杂精密仪器等缺点,不利于快速筛查 CRKP。因此,本研究旨在建立一个早期筛查 CRKP 的检测平台,以便及时进行有效的抗菌治疗,防止 CRKP 的广泛传播。我们将双 RPA 与 CRISPR/Cas12a 相结合,建立了肺炎克菌(Kp)rcsA 特异基因和 KPC 耐药基因的双重检测平台。我们建立了四种结果读取方法,包括荧光检测法(FD)、蓝光照射检测法(BLID)、紫外照射检测法(UID)和侧流检测条(LFTS)。对于 rcsA 基因,荧光检测法的 LOD 为 1 × 10 pg/μL,其他三种方法可检测出 1 × 101 pg/μL 的细菌 DNA。至于 KPC 基因,四种读出方法都能检测出 1 × 102 pg/μL 的细菌 DNA。在检测的 59 株临床菌株中,与培养法相比,RPA-CRISPR/Cas12a 双重检测 rcsA 的灵敏度、特异性和准确性均为 100%。与药敏试验相比,RPA-CRISPR/Cas12a 双重检测 KPC 的灵敏度为 85.71%,特异度为 100%,准确度为 94.92%。总之,事实证明我们的 RPA-CRISPR/Cas12a 双平台快速、精确、方便,可在实验室或医疗点有效检测 Kp 与 KPC。
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引用次数: 0
A millifluidic bioreactor allows the long term culture of primary lymphocytes or CD34+ hematopoietic cells while allowing the detection of tumorigenic expansion. 利用毫流体生物反应器可以长期培养原始淋巴细胞或 CD34+ 造血细胞,同时还能检测肿瘤扩增。
IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-02 eCollection Date: 2024-01-01 DOI: 10.3389/fbioe.2024.1388312
Paolo Ritter, Stefania Oliveto, Chiara Cordiglieri, Alessandra Fasciani, Christian Andrea Di Buduo, Lucrezia Della Volpe, Alberto Bocconi, Claudio Conci, Carolina Paula Miguel, Raffaella Di Micco, Alessandra Balduini, Manuela Teresa Raimondi, Stefano Biffo

Long-term culture of primary lymphocytes and hematopoietic stem and progenitor cells (HSPCs) is pivotal to their expansion and study. Furthermore, genetic engineering of the above-mentioned primary human cells has several safety needs, including the requirement of efficient in vitro assays for unwanted tumorigenic events. In this work, we tested and optimized the Miniaturized Optically Accessible Bioreactor (MOAB) platform. The MOAB consists of a millifluidic cell culture device with three optically-accessible culture chambers. Inside the MOAB, we inserted a silk-based framework that resembles some properties of the bone marrow environment and cultivated in this device either CD4+ T lymphocytes isolated from healthy donor buffy coat or cord blood-derived hematopoietic CD34+ cells. A fraction of these cells is viable for up to 3 months. Next, we tested the capability of the MOAB to detect tumorigenic events. Serial dilutions of engineered fluorescent tumor cells were mixed with either CD4+ or CD34+ primary cells, and their growth was followed. By this approach, we successfully detected as little as 100 tumorigenic cells mixed with 100,000 primary cells. We found that non-tumorigenic primary cells colonized the silk environment, whereas tumor cells, after an adaptation phase, expanded and entered the circulation. We conclude that the millifluidic platform allows the detection of rare tumorigenic events in the long-term culture of human cells.

原代淋巴细胞和造血干细胞及祖细胞(HSPCs)的长期培养对其扩增和研究至关重要。此外,对上述人类原代细胞进行基因工程有几种安全需求,包括需要高效的体外检测方法来检测不希望发生的致瘤事件。在这项工作中,我们测试并优化了微型可视生物反应器(MOAB)平台。MOAB 由带有三个光学可及培养室的毫流体细胞培养装置组成。我们在 MOAB 中插入了一个与骨髓环境某些特性相似的丝基框架,并在该装置中培养从健康供体水包衣中分离出来的 CD4+ T 淋巴细胞或脐带血造血 CD34+ 细胞。这些细胞中的一部分可存活长达 3 个月。接下来,我们测试了 MOAB 检测致瘤事件的能力。将连续稀释的工程荧光肿瘤细胞与 CD4+ 或 CD34+ 原始细胞混合,并跟踪其生长情况。通过这种方法,我们成功地检测到了100,000个原代细胞中混有100个致瘤细胞。我们发现,非致瘤性原代细胞在丝环境中定植,而肿瘤细胞在经过适应阶段后,扩大并进入血液循环。我们的结论是,毫流体平台可以在人类细胞的长期培养过程中检测到罕见的致瘤事件。
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引用次数: 0
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Frontiers in Bioengineering and Biotechnology
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