experimental design

实验设计
  • 文章类型: Journal Article
    在涉及小鼠等小型啮齿动物的研究中,尿液收集可能具有挑战性,因为实际的收集方法是焦虑的,并且限制了动物的福利,同时收集的尿液体积具有很大的变异性。为了改进目前的方法,最终减少对小鼠健康的影响,我们开发了一种创新的3D打印尿液收集设备(UCD)。这种两室UCD的形状适合于经典的饲养笼中,并允许通过自发排尿从两只装在自己笼子中的小鼠中收集尿液,而不会交叉污染,同时实现潜在的社交互动。我们使用UCD研究抗体介导的慢性肾脏疾病模型中与肾功能相关的尿参数的演变。总的来说,我们在这里报告了一种节省时间和负担得起的方法,用于收集大量未污染的尿液,我们认为与其他方法相比,这种方法可以改善动物的福利。
    Urine collection can be challenging in studies involving small rodents like mice, as the actual methods of collection are anxiogenic and constrain animal welfare while having high variability in the volume of urine collected. To improve the current methods and eventually reduce the impact on the well-being of mice, we developed an innovative 3D-printed urine collection device (UCD). This two-compartment UCD is shaped to fit in classical husbandry cages and allows urine collection by spontaneous urination from two mice housed in their own cage without cross-contamination while enabling potential social interactions. We used our UCD to study the evolution of urinary parameters related to renal functions in a model of antibody-mediated chronic kidney disease. Overall, we report here a time-saving and affordable method for urine collection providing a large amount of uncontaminated urine and which we believe may improve animal welfare in comparison with other methods.
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  • 文章类型: Journal Article
    本研究旨在使用实验设计来配制和评估用于控制乙酰唑胺(ACZ)递送的载有Eudragit纳米颗粒的水凝胶隐形眼镜。选择EudragitS-100来制备纳米颗粒。EudragitS100浓度的优化(X1),聚乙烯醇浓度(X2),通过应用中心复合实验设计尝试超声处理时间(X3)。纳米颗粒的平均尺寸(nm),从载有ACZ-EN的隐形眼镜中体外药物释放和药物浸出的百分比被认为是因变量。通过直接负载法制备了载有纳米颗粒的隐形眼镜并进行了表征。基于使用响应面方法开发的经过验证的二次多项式方程,选择了最佳检查点公式。优化的ACZ-EN配方呈现球形,尺寸为244.3nm,ζ电位为-13.2mV。发现纳米颗粒的包封效率为82.7±1.21%。使用自由基聚合技术成功制备了负载ACZ-EN的透明隐形眼镜。掺入隐形眼镜中的ACZ-EN表现出83.4±0.82%的溶胀和80.1±1.23%的透射率。当掺入隐形眼镜中时,ACZ-EN显示出药物的显著较低的突释,并且释放持续24小时。负载有隐形眼镜的ACZ-EN的灭菌制剂在兔眼中没有显示任何毒性迹象。由于ACZ-EN能够提供持续的药物释放并因此增强患者的依从性,因此可以将隐形眼镜中的ACZ-EN视为青光眼患者的潜在替代品。
    This study aims to formulate and evaluate Eudragit nanoparticles-laden hydrogel contact lenses for controlled delivery of acetazolamide (ACZ) using experimental design. Eudragit S-100 was selected for the preparation of nanoparticles. The optimization of Eudragit S100 concentration (X1), polyvinyl alcohol concentration (X2), and the sonication time (X3) was attempted by applying a central composite experimental design. Mean size of nanoparticles (nm), percent in vitro drug release and drug leaching from the ACZ-ENs laden contact lens were considered as dependent variables. Nanoparticles-laden contact lens was prepared through the direct loading method and characterized. Optimum check-point formulation was selected based on validated quadratic polynomial equations developed using response surface methodology. The optimized formulation of ACZ-ENs exhibited spherical shape with a size of 244.3 nm and a zeta potential of -13.2 mV. The entrapment efficiency of nanoparticles was found to be 82.7 ± 1.21%. Transparent contact lenses loaded ACZ-ENs were successfully prepared using the free radical polymerization technique. ACZ-ENs incorporated in contact lens exhibited a swelling of 83.4 ± 0.82% and transmittance of 80.1 ± 1.23%. ACZ-ENs showed a significantly lower burst release of the drug when incorporated in the contact lens and release was sustained over a period of 24 h. The sterilized formulation of ACZ-ENs laden contact lens did not show any sign of toxicity in rabbit eyes. ACZ-ENs incorporated in contact lens could be considered as a potential alternative in glaucoma patients due to their ability to provide sustained drug release and thus enhance patient compliance.
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  • 文章类型: Journal Article
    研究人员认为人类和其他动物共享认知能力是基于什么基础?我们认为,这种主张不是基于固有的,预先存在的相似性,而是通过两步过程出现,我们称之为“人为制造”。在初始阶段,由于需要测量和量化,因此忽略了人体研究中基于行动的策略和环境背景。因此,认知术语与我们应用它们的上下文脱节,人类分类认知术语被转化为广泛的解释性术语,假设是“物种中性”。第二阶段需要将这些广义的解释性术语翻译并应用于特定的非语言动物,以进一步掩盖动物与其他物种之间的差异。这里,再次,研究人员有选择地丢弃上下文信息,以方便与人类进行比较。为了限制人为制造,我们应该(重新)承认认知能力不是物种中性的,不能脱离具体行动,感知及其发生的背景。我们以记忆研究为例说明了我们关于人为制造的观点。本文是主题问题“运动中的思想:人工智能时代的具体认知”的一部分。
    On what basis do researchers posit that humans and other animals share cognitive capacities? We argue that such claims are not based on inherent, pre-existing similarities, but rather emerge through a two-step process, which we will call \'anthropofabrication\'. In the initial stage, embodied action-based strategies and environmental context in human studies are ignored owing to the need for measurement and quantification. Consequently, cognitive terms become disconnected from the context to which we apply them, and human classificatory cognitive terms are transformed into broad explanatory terms, assumed to be \'species-neutral\'. The second phase entails translating and applying these generalized explanatory terms to specific nonverbal animals in ways that serve to further cloak differences between animals and other species. Here, again, researchers selectively discard contextual information to facilitate the comparison with humans. To limit anthropofabrication, we should (re)acknowledge that cognitive abilities are not species-neutral and cannot be detached from embodied action, perception and their context of occurrence. We illustrate our points about anthropofabrication using the example of memory research. This article is part of the theme issue \'Minds in movement: embodied cognition in the age of artificial intelligence\'.
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  • 文章类型: Journal Article
    结论:计算信号去卷积的最新方法进展使得能够在更精细的细胞类型水平上进行大量转录组数据分析。通过反卷积,鉴定细胞类型特异性差异表达(csDE)基因在临床应用中引起越来越多的关注。然而,研究人员在实践中采用CSDE检测方法仍然面临许多困难,尤其是在他们的实验设计中.这里我们介绍柏树,首次实验设计和统计学分析工具在CSDE鉴定中的应用。该工具可以可靠地模拟纯化的细胞类型特异性(CTS)配置文件,细胞类型成分,生物和技术的变化,为批量RNA-seq卷积和反卷积提供高保真模拟器。赛柏树进行模拟,评估多个影响因素的影响,通过各种生物统计指标,帮助研究人员优化实验设计和进行功率分析。
    方法:cypress是位于https://bioparductor.org/packages/cypress/的开源R/Bioconductor软件包。
    背景:补充数据可在Bioinformatics在线获得。
    CONCLUSIONS: Recent methodology advances in computational signal deconvolution have enabled bulk transcriptome data analysis at a finer cell-type level. Through deconvolution, identifying cell-type-specific differentially expressed (csDE) genes is drawing increasing attention in clinical applications. However, researchers still face a number of difficulties in adopting csDE detection methods in practice, especially in their experimental design. Here we present cypress, the first experimental design and statistical power analysis tool in csDE identification. This tool can reliably model purified cell-type-specific (CTS) profiles, cell-type compositions, biological and technical variations, offering a high-fidelity simulator for bulk RNA-seq convolution and deconvolution. cypress conducts simulation and evaluates the impact of multiple influencing factors, by various biostatistical metrics, to help researchers optimize experimental design and conduct power analysis.
    METHODS: cypress is an open-source R/Bioconductor package at https://bioconductor.org/packages/cypress/.
    BACKGROUND: Supplementary data are available at Bioinformatics online.
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  • 文章类型: Journal Article
    基于化学计量学的方法被应用于从新原料中获得的生物炭中选择最有效的药物吸附剂。入侵植物(Ailanthusaltissima)的叶子。代表性的目标吸附物(阿替洛尔,扑热息痛,酮咯酸和四环素)是根据其理化性质选择的,以覆盖广泛的化学空间,这是通常的分析挑战。使用实验设计作为优化吸附系统性能的综合方法来研究它们的吸附,合理化程序,克服常见的缺点。在响应面设计中,选择了中心复合材料设计,因为它可以识别重要的实验因素(固液比,pH值,离子强度)及其相互作用,并允许选择最佳的实验条件,以最大限度地提高吸附性能。通过在500°C和800°C下热解制备生物炭(BC-500和BC-800),并在650°C和800°C下制备ZnCl2活化的生物炭(AcBC-650和AcBC-800)。FTIR光谱表明,在没有活化剂的情况下,提高热解温度会降低所有谱带的强度,虽然激活保留了官能团,如AcBC-650和AcBC-800的光谱所证明的。活化过程中的高温促进了有效表面积的发展,AcBC-800的最大值达到347m2g-1。AcBC-800被发现是最有效的吸附剂,阿替洛尔的去除效率为34.1、51.3、55.9和38.2%,扑热息痛,酮咯酸和四环素,分别。描述AcBC-800去除效率与实验因素之间关系的模型,显示令人满意的预测能力(预测R2>0.8),并且没有观察到显著的拟合缺乏。获得的结果,包括数学模型,吸附物和吸附剂的性质,明确表明,活性生物炭的吸附机制主要基于疏水相互作用,孔隙填充和氢键。
    A chemometrically based approach was applied to select the most efficient drug adsorbent among the biochars obtained from the novel feedstock, the leaves of the invasive plant (Ailanthus altissima). The representative target adsorbates (atenolol, paracetamol, ketorolac and tetracycline) were selected on the basis of their physicochemical properties to cover a wide chemical space, which is the usual analytical challenge. Their adsorption was investigated using design of experiments as a comprehensive approach to optimise the performance of the adsorption system, rationalise the procedure and overcome common drawbacks. Among the response surface designs, the central composite design was selected as it allows the identification of important experimental factors (solid-to-liquid ratio, pH, ionic strength) and their interactions, and allows the selection of optimal experimental conditions to maximise adsorption performance. The biochars were prepared by pyrolysis at 500 °C and 800 °C (BC-500 and BC-800) and the ZnCl2-activated biochars were prepared at 650 °C and 800 °C (AcBC-650 and AcBC-800). The FTIR spectra revealed that increasing the pyrolysis temperature without activator decreases the intensity of all bands, while activation preserves functional groups, as evidenced by the spectra of AcBC-650 and AcBC-800. High temperatures during activation promoted the development of an efficient surface area, with the maximum observed for AcBC-800 reaching 347 m2 g-1. AcBC-800 was found to be the most efficient adsorbent with removal efficiencies of 34.1, 51.3, 55.9 and 38.2 % for atenolol, paracetamol, ketorolac and tetracycline, respectively. The models describing the relationship between the removal efficiency of AcBC-800 and the experimental factors studied, showed satisfactory predictive ability (predicted R2 > 0.8) and no significant lack-of-fit was observed. The results obtained, including the mathematical models, the properties of the adsorbates and the adsorbents, clearly indicate that the adsorption mechanisms of activated biochars are mainly based on hydrophobic interactions, pore filling and hydrogen bonding.
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  • 文章类型: Journal Article
    紫外线辐射(UVR)是影响地球生命进化的普遍因素。UVR的环境水平介导关键的生物学功能,但也可以在广泛的生物体中引起严重的致死和亚致死效应。此外,UVR是其他环境因素对机体生理学影响的强大调节剂,如温度,疾病,毒理学和pH,在其他人中。这在全球变化的背景下至关重要,对于实验生物学家来说,理解多种应激源的影响是一个关键的挑战。生态生理学家很少提供UVR讨论或在实验设计中包括UVR,即使它与他们的学习系统直接相关。在这篇评论中,我们为实验生物学家提供了一个指导,以更好地理解,when,以及如何将UVR集成到实验设计中以提高其实验的生态真实感。
    Ultraviolet radiation (UVR) is a pervasive factor that has shaped the evolution of life on Earth. Ambient levels of UVR mediate key biological functions but can also cause severe lethal and sublethal effects in a wide range of organisms. Furthermore, UVR is a powerful modulator of the effects of other environmental factors on organismal physiology, such as temperature, disease, toxicology and pH, among others. This is critically important in the context of global change, where understanding the effects of multiple stressors is a key challenge for experimental biologists. Ecological physiologists rarely afford UVR discussion or include UVR in experimental design, even when it is directly relevant to their study system. In this Commentary, we provide a guide for experimental biologists to better understand if, when, and how UVR can be integrated into experimental designs to improve the ecological realism of their experiments.
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  • 文章类型: Journal Article
    在许多运动中,预期是表现的关键。根据定义,作为一个知觉-认知过程的预期旨在告知行动,并帮助运动员在时空压力下降低潜在的运动成本。预期研究一再因忽视行动而受到批评,并提出了在感知-行动耦合(PAC)条件下进行主要测试的必要性。据我们所知,然而,缺乏明确的标准来表征和定义PAC条件。这可能导致术语模糊,并可能使PAC条件和研究结果的解释和可比性复杂化。这里,我们提出了第一个建议,对PAC条件进行7级分类,并定义了刺激呈现和响应模式的维度。我们希望这种分类可以为预期研究中的研究计划和报告提供帮助。Further,在对球拍运动预期的回顾中,我们说明了PAC分类作为实验协议分析模板的潜在利用。对N=91篇文章中报道的N=115项研究的分析证实了代表性PAC条件的代表性不足,并且在该领域40多年的研究中,PAC方法几乎没有变化。我们讨论了这些发现的潜在原因,采用拟议的PAC分类的好处,并重申在预期研究中采取更多行动的呼吁。
    Anticipation is key to performance in many sports. By definition, anticipation as a perceptual-cognitive process is meant to inform action and help athletes reduce potential motor costs under spatiotemporal pressure. Anticipation research has repeatedly been criticized for neglecting action and raised the need for predominant testing under conditions of perception-action coupling (PAC). To the best of our knowledge, however, there is a lack of explicit criteria to characterize and define PAC conditions. This can lead to blurred terminology and may complicate interpretation and comparability of PAC conditions and results across studies. Here, we make a first proposal for a 7-level classification of PAC conditions with the defining dimensions of stimulus presentation and response mode. We hope this classification may constitute a helpful orientation for study planning and reporting in research on anticipation. Further, we illustrate the potential utilization of the PAC classification as a template for experimental protocol analysis in a review on anticipation in racket sports. Analysis of N = 115 studies reported in N = 91 articles confirms an underrepresentation of representative PAC conditions and reveals little change in PAC approaches over more than 40 years of research in that domain. We discuss potential reasons for these findings, the benefits of adopting the proposed PAC classification and reiterate the call for more action in anticipation research.
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  • 文章类型: Journal Article
    本研究旨在评估口服含有盐酸托莫西汀(ODMT)的快速微型片剂相对于盐酸托莫西汀(ATO)的常规胶囊制剂的功效。为了掩盖ATO的苦味,并使其更适合患有注意力缺陷多动障碍(ADHD)的人的儿科用药,合成了ATO与β-环糊精(β-CD)的包合物。将ODMT和常规胶囊ATO制剂口服给一组暴露于尼古丁的母鼠出生的ADHD幼鼠,促进体内疗效评估。行为分析,包括露天测试,新颖的物体识别测试,和巴恩斯迷宫测试,在治疗前后进行。结果表明,ODMT的配方,掺入ATO-β-CD包合物,显示了作为ATO胶囊形式的可行替代品的希望。最后,ATO-β-CD复合物和ODMT的制备利用析因实验设计,所述动物模型经受尼古丁诱导的活动过度,从而为正在开发的ODMT制剂提供独特的评价框架。
    The current study aimed to evaluate the efficacy of orally administered rapid mini-tablets containing atomoxetine hydrochloride (ODMT) relative to the conventional capsule formulation of atomoxetine hydrochloride (ATO). To mask the bitter taste of ATO and render it more palatable for pediatric administration in individuals with Attention Deficit Hyperactivity Disorder (ADHD), an inclusion complex of ATO with β-cyclodextrin (β-CD) was synthesized. The ODMT and conventional capsule ATO formulations were administered orally to a cohort of ADHD rat pups born to nicotine-exposed dams, facilitating an in vivo efficacy assessment. Behavioral assays, including the open field test, novel object recognition test, and Barnes maze test, were conducted pre- and post-administration of the therapeutics. The outcomes suggested that the ODMT formulation, incorporating ATO-β-CD inclusion complexes, shows promise as a viable alternative to the capsule form of ATO. Conclusively, the preparation of the ATO-β-CD complexes and ODMTs leveraged a factorial experimental design, with the animal model being subjected to nicotine-induced hyperactivity to provide a unique evaluative framework for the ODMT formulation under development.
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  • 文章类型: Journal Article
    标准参考材料(SRM)2806:液压油中的介质测试粉尘代表一系列由国家标准与技术研究所(NIST)认证的参考材料,用于校准液体光学(或自动)颗粒计数器,适用于广泛的工业,航空航天,和军事应用。该系列,包括SRM2806b,和SRM2806d,由IFTS为NIST制造,国际过滤器测试服务公司,在法国。材料接受认证的一个重要因素是瓶对瓶的均匀程度,由IFTS和NIST评估。开发了一种统计图形方法,该方法可提供即时的视觉和定量统计指标,以表征SRM。这种NIST开发的方法用于四项研究,以评估材料在其生产阶段和成品瓶装产品阶段的同质性。IFTS使用光学颗粒计数器进行测量以进行在线质量保证,并对成品400瓶系列的40瓶进行采样,以根据粒度分布确定均匀性。NIST还测定成品材料的粒度分布并进行显微镜检查以寻找悬浮液中可能的污染物。对两种材料(2806b和2806d)进行加速老化实验以验证它们的稳定性。
    Standard Reference Material (SRM) 2806: Medium Test Dust in Hydraulic Fluid represents a series of reference materials certified by the National Institute of Standards and Technology (NIST) used to calibrate liquid-borne optical (or automatic) particle counters applied in a wide range of industrial, aerospace, and military applications. The series, including SRM 2806b, and SRM 2806d, was manufactured for NIST by IFTS, Institut de la Filtration et des Techniques Séparatives International Filter Testing Services, in France. An important factor for the acceptance of the material for certification was the degree of bottle-to-bottle homogeneity, which was evaluated by both IFTS and NIST. A statistical graphics methodology was developed that provided immediate visual as well as quantitative statistical metrics with which to characterize the SRM. This NIST-developed approach was used in four studies to assess the homogeneity of the material during both its production stage and its finished bottled-product stage. IFTS performed measurements using an optical particle counter for on-line quality assurance and sampled 40 bottles of the finished 400 bottle series to determine homogeneity from the particle size distribution. NIST also determined the particle size distribution of the finished material and performed microscopy to look for possible contaminant material in the suspension. An accelerated aging experiment was conducted on both materials (2806b and 2806d) to verify their stability.
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  • 文章类型: Journal Article
    纳米结构脂质载体(NLC)已成为创新的药物递送系统,与其他基于脂质的载体相比,具有明显的优势,如脂质体和固体脂质纳米颗粒。苯佐卡因(BZC),使用中最古老的局部麻醉剂,通过假胆碱酯酶进行代谢,导致对氨基苯甲酸的形成,与长期使用BZC相关的过敏反应的病原体。为了减轻不良反应和提高生物利用度,BZC封装在NLC内。利用23阶乘设计,包含棕榈酸鲸蜡酯(固体脂质)的制剂,丙二醇单辛酸酯(液体脂质),并系统地制备了以PluronicF68为表面活性剂,随着固体/液体脂质质量比(60:40-80:20%)的变化,总脂质含量(15-25%),和BZC浓度(1-3%)。通过动态光散射对优化的配方进行表征,差示扫描量热法,拉曼成像,X射线衍射,小角度中子散射,纳米裂纹分析,透射电子显微镜(TEM)/低温TEM,提供对纳米颗粒结构和BZC掺入其脂质基质的见解。NLCBZC表现出显著的包封效率(%EE=96%)和在25°C下储存时的1年稳定性。在小鼠中进行的体外动力学研究和体内镇痛试验表明,NLCBZC有效地持续药物释放超过20小时,并将BZC的麻醉效果延长至18小时。因此,我们建议使用NLCBZC来减少苯佐卡因的有效麻醉浓度(从20到3%或更低),从而最大程度地减少局部施用这种麻醉剂后的过敏反应,潜在的,为疼痛管理中BZC给药的新途径铺平了道路。
    Nanostructured lipid carriers (NLC) have emerged as innovative drug delivery systems, offering distinct advantages over other lipid-based carriers, such as liposomes and solid lipid nanoparticles. Benzocaine (BZC), the oldest topical local anesthetic in use, undergoes metabolism by pseudocholinesterase, leading to the formation of p-aminobenzoic acid, a causative agent for allergic reactions associated with prolonged BZC usage. In order to mitigate adverse effects and enhance bioavailability, BZC was encapsulated within NLC. Utilizing a 23 factorial design, formulations comprising cetyl palmitate (solid lipid), propylene glycol monocaprylate (liquid lipid), and Pluronic F68 as surfactants were systematically prepared, with variations in the solid/liquid lipid mass ratios (60:40-80:20%), total lipid contents (15-25%), and BZC concentrations (1-3%). The optimized formulation underwent characterization by dynamic light scattering, differential scanning calorimetry, Raman imaging, X-ray diffraction, small-angle neutron scattering, nanotracking analysis, and transmission electron microscopy (TEM)/cryo-TEM, providing insights into the nanoparticle structure and the incorporation of BZC into its lipid matrix. NLCBZC exhibited a noteworthy encapsulation efficiency (%EE = 96%) and a 1 year stability when stored at 25 °C. In vitro kinetic studies and in vivo antinociceptive tests conducted in mice revealed that NLCBZC effectively sustained drug release for over 20 h and prolonged the anesthetic effect of BZC for up to 18 h. We therefore propose the use of NLCBZC to diminish the effective anesthetic concentration of benzocaine (from 20 to 3% or less), thus minimizing allergic reactions that follow the topical administration of this anesthetic and, potentially, paving the way for new routes of BZC administration in pain management.
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