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Sequencing, Fast and Slow: Profiling Microbiomes in Human Samples with Nanopore Sequencing 测序,快和慢:用纳米孔测序分析人类样品中的微生物组
Pub Date : 2023-08-17 DOI: 10.3390/applbiosci2030028
Yunseol Park, Jeesu Lee, Hyunjin Shim
Rapid and accurate pathogen identification is crucial in effectively combating infectious diseases. However, the current diagnostic tools for bacterial infections predominantly rely on century-old culture-based methods. Furthermore, recent research highlights the significance of host–microbe interactions within the host microbiota in influencing the outcome of infection episodes. As our understanding of science and medicine advances, there is a pressing need for innovative diagnostic methods that can identify pathogens and also rapidly and accurately profile the microbiome landscape in human samples. In clinical settings, such diagnostic tools will become a powerful predictive instrument in directing the diagnosis and prognosis of infectious diseases by providing comprehensive insights into the patient’s microbiota. Here, we explore the potential of long-read sequencing in profiling the microbiome landscape from various human samples in terms of speed and accuracy. Using nanopore sequencers, we generate native DNA sequences from saliva and stool samples rapidly, from which each long-read is basecalled in real-time to provide downstream analyses such as taxonomic classification and antimicrobial resistance through the built-in software (<12 h). Subsequently, we utilize the nanopore sequence data for in-depth analysis of each microbial species in terms of host–microbe interaction types and deep learning-based classification of unidentified reads. We find that the nanopore sequence data encompass complex information regarding the microbiome composition of the host and its microbial communities, and also shed light on the unexplored human mobilome including bacteriophages. In this study, we use two different systems of long-read sequencing to give insights into human microbiome samples in the ‘slow’ and ‘fast’ modes, which raises additional inquiries regarding the precision of this novel technology and the feasibility of extracting native DNA sequences from other human microbiomes.
快速准确的病原体鉴定是有效防治传染病的关键。然而,目前细菌感染的诊断工具主要依赖于已有百年历史的基于培养的方法。此外,最近的研究强调了宿主微生物群内宿主-微生物相互作用在影响感染事件结果中的重要性。随着我们对科学和医学的理解的进步,迫切需要创新的诊断方法,既能识别病原体,又能快速准确地描绘人类样本中的微生物群景观。在临床环境中,这种诊断工具将成为一种强大的预测工具,通过提供对患者微生物群的全面了解,指导传染病的诊断和预后。在这里,我们在速度和准确性方面探索了长读测序在分析各种人类样本微生物组景观方面的潜力。利用纳米孔测序仪,我们从唾液和粪便样本中快速生成天然DNA序列,通过内置软件(<12 h)实时调用每个长读段,以提供下游分析,如分类分类和抗微生物耐药性。随后,我们利用纳米孔序列数据对每个微生物物种进行深入分析,包括宿主-微生物相互作用类型和基于深度学习的未知读段分类。我们发现纳米孔序列数据包含了宿主及其微生物群落的微生物组组成的复杂信息,并揭示了包括噬菌体在内的未开发的人类移动组。在这项研究中,我们使用了两种不同的长读测序系统,以“慢”和“快”模式深入了解人类微生物组样本,这就提出了关于这种新技术的精度和从其他人类微生物组中提取天然DNA序列的可行性的额外疑问。
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引用次数: 0
Analysis of Adolescents’ Head to Shoulder Region during Tablet Use from Sagittal and Frontal RGB Images 矢状面和正面RGB图像对青少年使用片剂时头肩区域的分析
Pub Date : 2023-08-04 DOI: 10.3390/applbiosci2030027
Ivanna Kramer, S. Bauer
As schools go digital, the use of tablet computers is increasing. Concerns are raised that the extensive use of tablets and the associated bent-over posture may negatively affect the individual’s health. In order to analyse the possible effects of prolonged tablet use on physical health, a detailed analysis of the posture during tablet use is needed so that appropriate preventive measures can be taken to prevent degenerative changes. Therefore, the aim of this study was to measure and report the posture of 56 students while working with a tablet computer and compare it with an upright posture. Sagittal and frontal images were used for measurements of the subjects’ postures while seated, using the tablet, and in a neutral sitting position looking straight ahead. The body position during tablet use was recorded in two different user configurations: tablet flat on the table and tablet in individual freely chosen user configuration. After appropriate annotation of the data, the following parameters were evaluated in different planes. The craniovertebral angle (CVA), head tilt angle (HTA), and forward shoulder angle (FSA) are measurements that describe the extent to which the head bends forward and downward and how the shoulders are aligned in the sagittal plane. On the other hand, the head shoulder angle (HSA), lateral head tilt angle (LHTA), and trunk flexion angle (TFA) are angles measured in the frontal plane, which indicate the degree of head tilt and trunk bending to the right or left side. The measurement results clearly showed that the use of a tablet had a pronounced effect on the positions and rotations of the participants’ head, neck, and shoulders. This was evident through strong deviations observed in the angles measured between the sitting straight posture and the postures while using the tablet. For example, depending on the body posture class, the mean CVA values were 45.76° for straight sitting posture, 28.25° for holding the tablet individually posture, and 26.04° for the posture adopted while using a tablet placed flat on the table.
随着学校的数字化,平板电脑的使用也在增加。人们担心,大量使用片剂和相关的弯腰姿势可能会对个人健康产生负面影响。为了分析长期服用药片对身体健康可能产生的影响,需要详细分析服用药片时的姿势,以便采取适当的预防措施,防止退行性变化。因此,本研究的目的是测量和报告56名学生在使用平板电脑时的姿势,并将其与直立姿势进行比较。矢状面和额状面图像被用来测量受试者在坐着、使用平板电脑和处于直视前方的中立坐姿时的姿势。在平板电脑使用过程中,记录了两种不同的用户配置:平板电脑平放在桌子上和平板电脑在个人自由选择的用户配置。在对数据进行适当注释后,在不同平面上对以下参数进行评估。颅椎角(CVA)、头部倾斜角(HTA)和前肩角(FSA)是描述头部向前和向下弯曲的程度以及肩膀在矢状面上如何对齐的测量值。另一方面,头肩角(HSA)、头侧倾斜角(LHTA)和躯干弯曲角(TFA)是在正面测量的角度,表示头部倾斜和躯干向右或向左弯曲的程度。测量结果清楚地表明,使用平板电脑对参与者的头部、颈部和肩部的位置和旋转有明显的影响。这一点在使用平板电脑时观察到的坐直姿势和姿势之间的角度有明显的偏差。例如,根据身体姿势类别的不同,笔直坐姿的平均CVA值为45.76°,单独持片姿势的平均CVA值为28.25°,使用平板平放在桌子上的姿势的平均CVA值为26.04°。
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引用次数: 0
Effect of Doxapram, a K2p Channel Blocker, and pH on Heart Rate: Larval Drosophila Model Doxapram, K2p通道阻滞剂和pH对心率的影响:幼虫果蝇模型
Pub Date : 2023-08-03 DOI: 10.3390/applbiosci2030026
Elizabeth R. Elliott, Alaina C. Taul, Maya O. Abul-Khoudoud, Nicole Hensley, R. Cooper
Two-P-domain K+ (K2p) channels are responsible for maintaining the resting membrane potential. K2p channels have varied expression in healthy tissue, but they also change in cancerous or diseased states. The correlation and causation as regards the alteration of K2p channel expression are still being investigated. The compound doxapram seems to block K2p channels and depolarize cells. Using Drosophila, the increased expression of the ORK1 K2p channel in cardiac and skeletal muscle was investigated. The heart rate in larval Drosophila is very sensitive to pH, and since doxapram blocks a subset of the K2p channels that are known to be acid-sensitive, it was postulated that doxapram would affect heart rate. A pH change from 7.1 to 6.5 increased the rate, while that from 7.1 to 7.5 decreased the rate. An amount of 0.1 mM of doxapram had no effect, but 0.5 of mM depressed Drosophila heart rates within five minutes. Exposure to 5 mM of doxapram immediately decreased the rate. Lipopolysaccharides (LPSs) from Gram-negative bacteria acutely increased the rate. LPSs activate K2p channels in the skeletal muscle of larvae and are blocked by doxapram. LPSs slightly reduce depression in the rate induced by doxapram. The overexpression of K2p channels in the heart and skeletal muscle depressed the heart rate and heightened pH sensitivity. At larval neuromuscular junctions, the overexpression in skeletal muscle increases the frequency of spontaneous quantal events and produces a more negative resting membrane potential.
两个p域K+ (K2p)通道负责维持静息膜电位。K2p通道在健康组织中有不同的表达,但在癌变或病变状态下也会发生变化。K2p通道表达变化的相关性和原因仍在研究中。化合物doxapram似乎阻断了K2p通道并使细胞去极化。以果蝇为研究对象,研究了心肌和骨骼肌中ORK1 - K2p通道表达的增加。果蝇幼虫的心率对pH值非常敏感,由于多巴胺阻断了已知对酸敏感的K2p通道的一个子集,因此假设多巴胺会影响心率。pH值从7.1到6.5增加了反应速率,而从7.1到7.5则降低了反应速率。0.1 mM的doxapram没有效果,但0.5 mM的doxapram在5分钟内降低了果蝇的心率。暴露于5毫米的多巴胺会立即降低这一比率。来自革兰氏阴性菌的脂多糖(lps)急剧增加了这一比率。脂多糖激活幼体骨骼肌中的K2p通道,并被多巴胺阻断。脂多糖能轻微降低多巴胺诱导的抑郁率。心脏和骨骼肌中K2p通道的过度表达降低心率,提高pH敏感性。在幼体神经肌肉连接处,骨骼肌中的过表达增加了自发量子事件的频率,并产生更负的静息膜电位。
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引用次数: 0
Automation of Cluster Extraction in Fundus Autofluorescence Images of Geographic Atrophy 地理萎缩眼底自荧光图像聚类提取的自动化
Pub Date : 2023-07-21 DOI: 10.3390/applbiosci2030025
J. Arslan, K. Benke
The build-up of lipofuscin—an age-associated biomarker referred to as hyperfluorescence—is considered a precursor in the progression of geographic atrophy (GA). Prior studies have attempted to classify hyperfluorescent regions to explain varying rates of GA progression. In this study, digital image processing and unsupervised learning were used to (1) completely automate the extraction of hyperfluorescent regions from images, and (2) evaluate prospective patterns and groupings of hyperfluorescent areas associated with varying levels of GA progression. Patterns were determined by clustering methods, such as k-Means, and performance was evaluated using metrics such as the Silhouette Coefficient (SC), the Davies–Bouldin Index (DBI), and the Calinski–Harabasz Index (CHI). Automated extraction of hyperfluorescent regions was carried out using pseudocoloring techniques. The approach revealed three distinct types of hyperfluorescence based on color intensity changes: early-stage hyperfluorescence, intermediate-stage hyperfluorescence, and late-stage hyperfluorescence, with the early and late stages having three additional subclassifications that could explain varying levels of GA progression. The performance metrics for early-stage hyperfluorescence were SC = 0.597, DBI = 0.915, and CHI = 186.989. For late-stage hyperfluorescence, SC = 0.593, DBI = 1.013, and CHI = 217.325. No meaningful subclusters were identified for the intermediate-stage hyperfluorescence, possibly because it is a transitional phase of hyperfluorescence progression.
脂褐素(一种与年龄相关的生物标志物,称为高荧光)的积累被认为是地理萎缩(GA)进展的前兆。先前的研究试图对高荧光区进行分类,以解释GA进展的不同速率。在本研究中,使用数字图像处理和无监督学习来(1)完全自动化地从图像中提取高荧光区域,(2)评估与不同GA进展水平相关的高荧光区域的预期模式和分组。通过聚类方法(如k-Means)确定模式,并使用廓形系数(SC)、Davies-Bouldin指数(DBI)和Calinski-Harabasz指数(CHI)等指标评估绩效。利用伪着色技术自动提取高荧光区域。该方法根据颜色强度变化揭示了三种不同类型的高荧光:早期高荧光、中期高荧光和晚期高荧光,早期和晚期有三个额外的亚分类,可以解释不同程度的GA进展。早期高荧光性能指标SC = 0.597, DBI = 0.915, CHI = 186.989。晚期高荧光SC = 0.593, DBI = 1.013, CHI = 217.325。中期高荧光没有确定有意义的亚群,可能是因为它是高荧光进展的过渡阶段。
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引用次数: 0
Review of Partial Hybrids between Herbaceous Medicago sativa and Woody Medicago arborea and Their Potential Role in Alfalfa Improvement 草本紫花苜蓿与木本紫花苜蓿部分杂交研究进展及其在苜蓿改良中的潜在作用
Pub Date : 2023-07-13 DOI: 10.3390/applbiosci2030024
J. Irwin, E. Bingham
Medicago sativa (2n = 4x = 32) and M. arborea (2n = 4x = 32) were thought to be reproductively isolated until hybrids (Alborea) were produced by sexual reproduction for the first time in 2003 in Wisconsin. The hybrids were asymmetric, at or near 2n = 4x = 32, and with a predominance of the alfalfa genome. Only M. sativa seed parents with reproductive abnormalities, including unreduced eggs, have produced hybrids; where M. arborea has been used as the seed parent, no hybrids have resulted. Pedigree selection within derivatives of the two original M. sativa seed parents (MB and M8) has been successful in increasing the frequency of hybrids produced. While Alborea individuals more closely resemble M. sativa, a number of M. arborea-specific traits have been observed across different hybrid individuals. These include single-coil flat pods, large seeds, yellow flowers, indeterminate growth, a minimal crown, lodging, frost resistance, and anthracnose resistance. These M. arborea traits have the potential to restructure alfalfa to increase its versatility and utilisation. There is emerging evidence from North and South America and Australia that some Alborea selections have the capacity to complement adapted alfalfa cultivars for yield. Work is continuing to introgress M. arborea traits of value into alfalfa.
苜蓿(2n = 4x = 32)和树楸(2n = 4x = 32)一直被认为是生殖分离的,直到2003年在威斯康星州首次通过有性繁殖产生了杂种植物(Alborea)。杂交种不对称,在2n = 4x = 32附近,具有苜蓿基因组优势。只有具有生殖异常(包括未减少的卵子)的sativa种子亲本才能产生杂交种;在用木本松作为种亲本的地方,没有杂交的结果。对两种原始sativa种子亲本(MB和M8)的衍生物进行系谱选择,成功地提高了杂交种的产生频率。虽然白桦个体更接近于苜蓿,但在不同的杂交个体中观察到许多白桦特有的性状。这些包括单圈扁平的豆荚,大的种子,黄色的花,不确定的生长,最小的冠,倒伏,抗冻性,和抗炭疽病。这些特征有可能重组紫花苜蓿,以增加其多功能性和利用率。来自北美、南美和澳大利亚的新证据表明,一些桤木选种有能力补充适应的苜蓿品种的产量。将有价值的木蠹性状引入紫花苜蓿的工作仍在继续。
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引用次数: 0
Wine Grapes Ripening: A Review on Climate Effect and Analytical Approach to Increase Wine Quality 酿酒葡萄成熟:气候影响及提高葡萄酒品质的分析方法综述
Pub Date : 2023-07-10 DOI: 10.3390/applbiosci2030023
M. Rouxinol, M. Martins, J. M. Barroso, A. Rato
Red wine grapes have an important impact on the economy of many regions, both for wine quality and for their richness in phenolic compounds, which have many health benefits. Climate has been changing substantially in the last years, which affects greatly grape polyphenolic composition and wine quality. In this review, we will unveil the importance of climate in grape development, both physically and chemically, the different methodologies used to evaluate grape quality, the interesting new approaches using NIR spectroscopy, and the functional properties of grapes and red wine, due to their high phenolic content. Climate has an impact in the development of phenolic compounds in grapes, namely in the anthocyanins biosynthesis. The phenolic chemical composition changes during maturation, therefore, it is essential to keep on track the accumulation of these key compounds. This information is crucial to help producers choose the best harvest date since specific compounds like polyphenols are responsible for the color, taste, and mouthfeel of wines, which directly affects wine quality. The usage of different methodologies to assess quality parameters in grapes and wine, can be used to provide essential information to create the chemical profile of each variety to develop calibration methods. NIR spectroscopy seems to be a reliable method to be used in vineyards during grape maturation to provide real time information on quality parameters to producers since many reliable calibration models have been developed over time.
红葡萄酒葡萄对许多地区的经济都有重要影响,无论是葡萄酒质量还是其丰富的酚类化合物,都有许多健康益处。在过去的几年里,气候发生了很大的变化,这极大地影响了葡萄多酚成分和葡萄酒的品质。在这篇综述中,我们将揭示气候在葡萄发育中的重要性,包括物理和化学,用于评估葡萄品质的不同方法,利用近红外光谱的有趣的新方法,以及葡萄和红葡萄酒由于其高酚含量而具有的功能特性。气候影响葡萄中酚类化合物的发育,即花青素的生物合成。在成熟过程中,酚类化合物的化学成分发生了变化,因此,跟踪这些关键化合物的积累是必要的。这些信息对于帮助生产者选择最佳收获日期至关重要,因为多酚等特定化合物负责葡萄酒的颜色,味道和口感,直接影响葡萄酒的质量。使用不同的方法来评估葡萄和葡萄酒的质量参数,可以用来提供必要的信息,以创建每个品种的化学特征,以制定校准方法。随着时间的推移,许多可靠的校准模型已经开发出来,因此在葡萄成熟过程中,近红外光谱似乎是一种可靠的方法,可以为生产者提供质量参数的实时信息。
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引用次数: 0
Exploring the Synergistic Impacts of Cover Crops and Fertilization on Soil Microbial Metabolic Diversity in Dryland Soybean Production Systems Using Biolog EcoPlates 利用生物生态板研究覆盖作物和施肥对旱地大豆生产系统土壤微生物代谢多样性的协同影响
Pub Date : 2023-07-06 DOI: 10.3390/applbiosci2030022
Durga P. M. Chinthalapudi, Sapna Pokhrel, W. Kingery, M. Shankle, Shankar Ganapathi Shanmugam
The metabolic diversity of soil microbiota embodies diverse functional capabilities that support ecosystem resilience, driving essential biogeochemical processes and facilitating the optimization of sustainable agricultural systems. Integrating cover crops into agricultural systems cultivates a diverse array of metabolic activities among soil microbes, synergistically enhancing ecosystem services and bolstering soil health for sustainable and productive farming practices. In an effort to gain deeper insights and expand our knowledge, we conducted a study examining the effects of cover crops and fertilizer sources, thereby shedding light on their combined impacts on the metabolic activity dynamics of soil microbial communities. In this investigation, we employed a split-plot design with two factors: (a) cover crop with three solo cover crop species—Cereal rye (Secale cereale), wheat (Triticum aestivum), hairy vetch (Vicia villosa), and one mixture of mustard (Brassica rapa) and cereal rye (Secale cereale) (CC-mix), (b) Fertilizer source includes poultry litter, chemical fertilizer, and no-fertilizer treatments. We assessed the metabolic potential of soil microbiota by using carbon substrates utilizing Biolog EcoPlates. The findings revealed that the plots with CC-mix treatment exhibited greater metabolic diversity compared to the other treatments, while among the fertilizer sources, poultry litter demonstrated higher metabolic activity. Furthermore, both treatment factors predominantly metabolized carbohydrates and polymers compared to other carbon substrate categories. The principal component analysis accounted for 46.4% of the variance, collectively represented by PC1 and PC2, emphasizing the substantial contributions of carbohydrates, amino acids, and carboxylic acids to the observed metabolic diversity. Canonical correspondence analysis revealed that pH had positively correlated with microbial functional diversity, whereas total carbon (TC), total nitrogen (TN), and water-stable aggregates (WSA) showed a negative correlation. In conclusion, cover cropping and type of fertilizer source had a notable impact on soil microbial functional diversity, with the cover crop mixture exhibiting a more pronounced influence than the individual cover crop treatments.
土壤微生物群的代谢多样性体现了支持生态系统恢复力、驱动基本生物地球化学过程和促进可持续农业系统优化的多种功能能力。将覆盖作物纳入农业系统可促进土壤微生物的多种代谢活动,协同增强生态系统服务,促进土壤健康,促进可持续和生产性耕作方式。为了获得更深入的认识和扩展我们的知识,我们进行了一项研究,考察了覆盖作物和肥料来源的影响,从而揭示了它们对土壤微生物群落代谢活性动态的综合影响。在本研究中,我们采用了两个因素的分块设计:(a)覆盖作物为3种单独覆盖作物,即黑麦(Secale cereale)、小麦(Triticum aestivum)、野豌豆(Vicia villosa)和芥菜(Brassica rapa)和黑麦(Secale cereale)的混合(c -mix); (b)肥料来源包括家禽凋落物、化肥和无肥处理。利用Biolog EcoPlates利用碳基质对土壤微生物群的代谢潜力进行了评价。结果表明,与其他肥料处理相比,CC-mix处理具有更大的代谢多样性,而在肥料来源中,家禽粪便的代谢活性更高。此外,与其他碳底物类别相比,这两种处理因素主要代谢碳水化合物和聚合物。主成分分析占46.4%的方差,由PC1和PC2共同代表,强调碳水化合物、氨基酸和羧酸对观察到的代谢多样性的重要贡献。典型对应分析表明,pH值与微生物功能多样性呈正相关,而总碳(TC)、总氮(TN)和水稳性团聚体(WSA)呈负相关。综上所述,覆盖作物和肥料来源类型对土壤微生物功能多样性有显著影响,覆盖作物混作处理对土壤微生物功能多样性的影响比单种覆盖作物处理更显著。
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引用次数: 0
Trichoderma species have potential in suppressing Fusarium oxysporum f.sp cubense infecting banana in Kenya 木霉对肯尼亚香蕉尖孢镰刀菌有潜在的抑制作用
Pub Date : 2023-06-30 DOI: 10.35759/jabs.186.10
Samuel Musime Malaka, D. M. Thuranira, M. Mwangi, S. Nchore, Hudson Lubabali, Sylvia W Kuria, Daniel Omingo Omari, C. Gathambiri
Objectives: This study examined the effectiveness of carbendazim, Trichoderma harzianum, T. asperellum, T. viride, and a combination of the carbendazim and the bio-fungicides for the management of Foc both under in vitro and greenhouse conditions. Methodology and Results: The poisoned food technique was used in the in vitro tests. Fungal radial growth was recorded daily for seven days. In the greenhouse, the test products and Foc were applied directly into the soil, disease severity data was recorded at seven day intervals for 98 days using a scale of 1-5. Data on plant biomass was recorded at the end of the experiment (98 days). ANOVA was used to analyze data on fungal radial growth, disease severity, and plant biomass. The Student-Newman-Keuls (SNK) test was used to compare the results at P≤0.05. For the in vitro test, all treatments significantly (P<0.05) suppressed the growth of Foc compared to the untreated control. In the greenhouse trials a significant difference (P<0.05) in external/yellowing symptoms was observed but vascular discoloration was not different. Conclusions and application of findings: This study provides insights into the performance of bio fungicides and carbendazim in managing Panama disease. Keywords: Banana, Fusarium oxysporum f.sp. cubense, , Panama disease, Trichoderma sp.
目的:本研究考察了多菌灵、哈茨木霉、曲霉、绿霉以及多菌灵与生物杀菌剂联合使用在体外和温室条件下对Foc的治理效果。方法与结果:采用中毒食品法进行体外试验。连续7天每天记录真菌径向生长情况。在温室内,试验产品和Foc直接施用于土壤中,每隔7天记录病害严重程度数据,连续98 d,按1-5分进行。试验结束时(98 d)记录植物生物量数据。采用方差分析分析真菌径向生长、疾病严重程度和植物生物量的数据。采用Student-Newman-Keuls (SNK)检验比较,P≤0.05。在体外试验中,与未处理对照组相比,各处理均显著(P<0.05)抑制了Foc的生长。在温室试验中,外黄化症状有显著性差异(P<0.05),但血管变色无显著性差异。结论和研究结果的应用:本研究为生物杀菌剂和多菌灵在管理巴拿马病中的性能提供了见解。关键词:香蕉;尖孢镰刀菌;巴拿马病,木霉属。
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引用次数: 0
Comparison of Culture Media for In Vitro Expansion of Oral Epithelial Keratinocytes 口腔上皮角质形成细胞体外扩增培养基的比较
Pub Date : 2023-06-16 DOI: 10.3390/applbiosci2020021
Giancarlo A. Cuadra, Abrar Shamim, Raivat Shah, Joey Morgan, D. Palazzolo
Background: Expansion of OKF6/TERT-2 oral epithelial cells in vitro is important for studying the molecular biology of disease and pathology affecting the oral cavity. Keratinocyte serum-free medium (KSFM) is the medium of choice for this cell line. This study compares three media for OKF6/TERT-2 cultures: KSFM, Dulbecco’s Modified Eagle Medium/Nutrient Mixture of Hams F-12 (DMEM/F12), and a composite medium comprised of DMEM/F-12 and KSFM (1:1 v/v), referred to as DFK. The toxicological effects of electronic cigarette liquids (e-liquids) on OKF6/TERT-2 cells cultured in these media were also compared. Methods: Cells were cultured in KSFM, DMEM/F12, or DFK, and cellular morphology, growth, wound healing and the gene expression of mucins and tight junctions were evaluated. Additionally, cytotoxicity was determined after e-liquid exposures. Results: Switching from KSFM to DMEM/F12 or DFK 24 h post-seeding leads to typical cellular morphologies, and these cultures reach confluency faster than those in KSFM. Wound-healing recovery occurred fastest in DFK. Except for claudin-1, there is no difference in expression of the other genes tested. Additionally, e-liquid cytotoxicity appears to be amplified in DFK cultures. Conclusions: DMEM/F12 and DFK are alternative media for OKF6/TERT-2 cell culture to study the molecular biology of disease and pathology, provided cells are initially seeded in KSFM.
背景:OKF6/TERT-2口腔上皮细胞的体外扩增对研究口腔疾病和病理的分子生物学具有重要意义。无血清角质细胞培养基(KSFM)是该细胞系的培养基选择。本研究比较了OKF6/TERT-2培养的三种培养基:KSFM, Dulbecco 's Modified Eagle培养基/火腿F-12营养混合物(DMEM/F12),以及由DMEM/F-12和KSFM (1:1 v/v)组成的复合培养基,称为DFK。并比较了电子烟液对OKF6/TERT-2细胞的毒理学影响。方法:分别在KSFM、DMEM/F12、DFK中培养细胞,观察细胞形态、生长、创面愈合、粘蛋白和紧密连接基因表达。此外,还测定了电子烟液暴露后的细胞毒性。结果:播种24 h后,从KSFM切换到DMEM/F12或DFK可以获得典型的细胞形态,并且这些培养比KSFM更快地达到融合。DFK组伤口愈合速度最快。除claudin-1外,其他基因的表达均无差异。此外,电子液体的细胞毒性似乎在DFK培养中被放大。结论:DMEM/F12和DFK是OKF6/TERT-2细胞培养研究疾病和病理分子生物学的替代培养基,前提是细胞最初播种在KSFM中。
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引用次数: 0
An Event-Driven Architecture for Genomics-Based Diagnostic Data Processing 基于基因组学的诊断数据处理的事件驱动架构
Pub Date : 2023-06-02 DOI: 10.3390/applbiosci2020020
Thomas Krause, Mike Zickfeld, Sebastian Bruchhaus, Thoralf Reis, M. X. Bornschlegl, P. Buono, Michael Kramer, Paul Mc Kevitt, M. Hemmje
Genomics-based diagnostic data (GBDD) are becoming increasingly important for laboratory diagnostics. Due to the large quantity of data and their heterogeneity, GBDD poses a big data challenge. Current analysis tools for GBDD are primarily designed for research and do not meet the requirements of laboratory diagnostics for automation, reliability, transparency, reproducibility, robustness, and accessibility. This makes it difficult for laboratories to use these tools in tests that need to be validated according to regulatory frameworks and to execute tests in a time- and cost-efficient manner. In order to better address these requirements, we propose an event-driven workflow-based architecture as the basis for a processing platform that is highly scalable using container technologies and microservices. A prototype implementation of this approach, called GenomicInsights, has been developed and evaluated to demonstrate its feasibility and suitability for laboratory diagnostics.
基于基因组学的诊断数据(GBDD)对实验室诊断越来越重要。由于数据量大,数据的异质性,GBDD面临着大数据的挑战。目前GBDD的分析工具主要是为研究而设计的,不能满足实验室诊断对自动化、可靠性、透明度、可重复性、稳健性和可及性的要求。这使得实验室难以在需要根据监管框架进行验证的测试中使用这些工具,也难以以时间和成本效益高的方式执行测试。为了更好地满足这些需求,我们提出了一个基于事件驱动工作流的架构,作为使用容器技术和微服务的高度可伸缩处理平台的基础。已经开发并评估了这种方法的原型实现,称为GenomicInsights,以证明其在实验室诊断中的可行性和适用性。
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引用次数: 0
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Journal of Applied Biosciences
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