Microscope imaging

  • 文章类型: Journal Article
    我们在此报告非格司亭产品Neupogen®和非格司亭制剂缓冲液在体外与人血浆混合时诱导聚集体形成,与人的血液混合时形成大的膜红细胞聚集体,类似于pegfilgrastim和pegfilgrastim缓冲液[T.Arvinte,E.Poirier,N.Ersayin,G.Darpin,A.Cudd,J.Dowd,S.Brokx,pegfilgrastim鼻祖制剂缓冲液和pegfilgrastim产品在体外诱导人血浆和人血液的聚集,欧尔.J.Pharmaceut。生物制药。(2023),doi:10.1016/j.ejpb.2023.10.019]。数据确定了filgrastim缓冲液(在filgrastim和pegfilgrastim产品中几乎相同)作为人血浆和血液聚集的主要驱动因素。动力学实验表明,由欧盟制造的非格司亭产品和美国制造的非格司亭产品诱导的血浆聚集程度存在差异。观察到非格司亭诱导的血浆聚集中的人类供体变异性。为了研究非格司亭蛋白的聚乙二醇化对血浆聚集的影响,我们比较了相同蛋白质浓度的非格司亭(Neupogen®)与pegfilgrastim(Neulasta®)溶液。这些数据表明,PEG化具有在一定程度上抑制血浆聚集的有益效果。有趣的是,与缓冲液相比,非格司亭缓冲液中的20kDa聚乙二醇诱导了更多的血浆聚集,类似于pegfilgrastim诱导的聚集。对于静脉输注非格司亭溶液(300µg/ml,仅小瓶)可以在5%葡萄糖中从300μg/ml的浓度稀释至5μg/ml。人血浆的聚集也通过在5%葡萄糖中稀释至50μg/ml的非格司亭溶液诱导,15微克/毫升和5微克/毫升非格司亭,以及类似地在5%葡萄糖中稀释的非格司亭缓冲液(1:6、1:20和1:60稀释)。这些数据表明,用于患者静脉内给药的非格司亭溶液在体外诱导人血浆聚集。这种聚集现象可能与非格司亭治疗的已知输注副作用有关。
    We herein report that filgrastim product Neupogen® and the filgrastim formulation buffer induced aggregate formation when mixed in vitro with human plasma, and formation of large membranous erythrocyte aggregates when mixed with human blood, similar to the aggregation induced by pegfilgrastim and by pegfilgrastim buffer [T. Arvinte, E. Poirier, N. Ersayin, G. Darpin, A. Cudd, J. Dowd, S. Brokx, Aggregation of human plasma and of human blood induced in vitro by pegfilgrastim originator formulation buffer and pegfilgrastim products, Eur. J. Pharmaceut. Biopharmaceut. (2023), doi: 10.1016/j.ejpb.2023.10.019]. The data identify the filgrastim buffer (which is practically the same in filgrastim and pegfilgrastim products) as the main driver of human plasma and blood aggregation. Kinetic experiments showed differences in the extent of plasma aggregation induced by a filgrastim product manufactured in EU and one manufactured in USA. Human donor variability in the plasma aggregation induced by filgrastim was observed. To study the effect of PEGylation of the filgrastim protein on plasma aggregation we compared filgrastim (Neupogen®) with pegfilgrastim (Neulasta®) solutions at the same protein concentration. These data show that PEGylation has a beneficial effect in inhibiting to an extent plasma aggregation. Interestingly, 20 kDa polyethylene glycol in the filgrastim buffer induced more plasma aggregation compared to the buffer, similar to the aggregation induced by pegfilgrastim. For intravenous infusion filgrastim solutions (300 µg/ml, vials only) may be diluted in 5 % dextrose from a concentration of 300 µg/ml to 5 µg/ml. Aggregation of human plasma was also induced by filgrastim solutions diluted in 5 % dextrose to 50 µg/ml, 15 µg/ml and 5 µg/ml filgrastim, as well as by the filgrastim buffer similarly diluted in 5 % dextrose (1:6, 1:20 and 1:60 dilution). These data show that filgrastim solutions used for intravenous administration in patients induce human plasma aggregation in vitro. Such aggregation phenomena may be related to known infusion side effects of filgrastim therapy.
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  • 文章类型: Journal Article
    基因调控网络现在处于精准生物学的前沿,这可以帮助研究人员更好地了解基因和调控元件如何相互作用以控制细胞基因表达,在生物学研究中提供了更有前途的分子机制。基因和调控元件之间的相互作用涉及不同的启动子,增强器,转录因子,消音器,绝缘子,和远程监管元素,它们以时空方式发生在10µm的核上。这样,三维染色质构象和结构生物学对于解释生物效应和基因调控网络至关重要。在审查中,我们简要总结了三维染色质构象的最新过程,显微成像,和生物信息学,我们对这三个方面提出了展望和未来方向。
    Gene regulatory networks are now at the forefront of precision biology, which can help researchers better understand how genes and regulatory elements interact to control cellular gene expression, offering a more promising molecular mechanism in biological research. Interactions between the genes and regulatory elements involve different promoters, enhancers, transcription factors, silencers, insulators, and long-range regulatory elements, which occur at a ∼10 µm nucleus in a spatiotemporal manner. In this way, three-dimensional chromatin conformation and structural biology are critical for interpreting the biological effects and the gene regulatory networks. In the review, we have briefly summarized the latest processes in three-dimensional chromatin conformation, microscopic imaging, and bioinformatics, and we have presented the outlook and future directions for these three aspects.
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  • 文章类型: Journal Article
    显微照片比较在生物科学中仍然有用。这项技术为研究人员提供了实验条件的快速快照。但有时两种情况的比较依赖于研究人员的眼睛来得出结论。我们的生物图像分析,统计,和比较(BASIN)软件提供了一个客观和可重复的比较,利用推理统计来桥接图像数据与其他模式。用户可以访问基于机器学习的对象分割。BASIN提供了几个数据点,如图像\'对象计数,强度,和地区。也可以进行假设检验。为了提高盆地的可访问性,我们使用RShiny实现了它,并提供了在线和离线版本。我们使用BASIN处理了涉及五个生物科学主题的498个图像对。我们的框架有57%的时间支持直接索赔或外推。对分析结果进行手动整理,以确定BASIN的准确性,该准确性显示为78%。此外,每个BASIN版本的初始版本显示平均82%的FAIR合规性得分。
    Micrograph comparison remains useful in bioscience. This technology provides researchers with a quick snapshot of experimental conditions. But sometimes a two- condition comparison relies on researchers\' eyes to draw conclusions. Our Bioimage Analysis, Statistic, and Comparison (BASIN) software provides an objective and reproducible comparison leveraging inferential statistics to bridge image data with other modalities. Users have access to machine learning-based object segmentation. BASIN provides several data points such as images\' object counts, intensities, and areas. Hypothesis testing may also be performed. To improve BASIN\'s accessibility, we implemented it using R Shiny and provided both an online and offline version. We used BASIN to process 498 image pairs involving five bioscience topics. Our framework supported either direct claims or extrapolations 57% of the time. Analysis results were manually curated to determine BASIN\'s accuracy which was shown to be 78%. Additionally, each BASIN version\'s initial release shows an average 82% FAIR compliance score.
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  • 文章类型: Journal Article
    We present here a novel multi-parametric approach for the characterization of multiple cellular features, using images acquired by high-throughput and high-definition light microscopy. We specifically used this approach for deep and unbiased analysis of the effects of a drug library on five cultured cell lines. The presented method enables the acquisition and analysis of millions of images, of treated and control cells, followed by an automated identification of drugs inducing strong responses, evaluating the median effect concentrations and those cellular properties that are most highly affected by the drug. The tools described here provide standardized quantification of multiple attributes for systems level dissection of complex functions in normal and diseased cells, using multiple perturbations. Such analysis of cells, derived from pathological samples, may help in the diagnosis and follow-up of treatment in patients.
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  • 文章类型: Journal Article
    在拟南芥中,绞股蓝,花的内圈,是女性的生殖部分。许多对受精很重要的组织,如柱头,风格,发射道,胎盘,胚珠,和隔膜,包括中间域,起源于心皮边缘分生组织。在绞股蓝发育过程中,发生隔膜融合,组织不断形成,为成功的授粉和受精做准备。在绞股蓝发育过程中,发生细胞壁修饰,最重要的是传输道的形成,对生殖能力有很大影响,因为它有助于花粉管的生长和通过卵巢的运动。在这项研究中,使用经典染色方法的组合,荧光染料,和间接免疫定位,我们分析了在雌蕊发育过程中伴随内侧结构域形成的细胞壁组成和修饰。我们检测到多糖分布随时间的协调变化,在传输道形成之前的细胞壁修饰,粘液物质在传播道形成过程中增加,甘露聚糖分布减少。此外,我们还检测到隔膜融合过程中脂质分布的变化.正确的细胞壁组成和修饰对于心皮的生殖器后融合(隔膜融合)和传输道的形成很重要,因为这些组织影响植物的繁殖能力。
    In Arabidopsis, the gynoecium, the inner whorl of the flower, is the female reproductive part. Many tissues important for fertilization such as the stigma, style, transmitting tract, placenta, ovules, and septum, comprising the medial domain, arise from the carpel margin meristem. During gynoecium development, septum fusion occurs and tissues form continuously to prepare for a successful pollination and fertilization. During gynoecium development, cell wall modifications take place and one of the most important is the formation of the transmitting tract, having a great impact on reproductive competence because it facilitates pollen tube growth and movement through the ovary. In this study, using a combination of classical staining methods, fluorescent dyes, and indirect immunolocalization, we analyzed cell wall composition and modifications accompanying medial domain formation during gynoecium development. We detected coordinated changes in polysaccharide distribution through time, cell wall modifications preceding the formation of the transmitting tract, mucosubstances increase during transmitting tract formation, and a decrease of mannan distribution. Furthermore, we also detected changes in lipid distribution during septum fusion. Proper cell wall composition and modifications are important for postgenital fusion of the carpel (septum fusion) and transmitting tract formation, because these tissues affect plant reproductive competence.
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