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Mouse aversion to induction with isoflurane using the drop method. 使用滴注法诱导小鼠厌恶异氟醚。
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-04 DOI: 10.1177/00236772241262119
Maya J Bodnar, I Joanna Makowska, Courtney T Boyd, Catherine A Schuppli, Daniel M Weary

Isoflurane anesthesia prior to carbon dioxide euthanasia is recognized as a refinement by many guidelines. Facilities lacking access to a vaporizer can use the "drop" method, whereby liquid anesthetic is introduced into an induction chamber. Knowing the least aversive concentration of isoflurane is critical. Previous work has demonstrated that isoflurane administered with the drop method at a concentration of 5% is aversive to mice. Other work has shown that lower concentrations (1.7% to 3.7%) of isoflurane can be used to anesthetize mice with the drop method, but aversion to these concentrations has not been tested. We assessed aversion to these lower isoflurane concentrations administered with the drop method, using a conditioned place aversion (CPA) paradigm. Female C57BL/6J (OT-1) mice (n = 28) were randomly allocated to one of three isoflurane concentrations: 1.7%, 2.7%, and 3.7%. Mice were acclimated to a light-dark apparatus. Prior to and following dark (+ isoflurane) and light chamber conditioning sessions, mice underwent an initial and final preference assessment; the change in the duration spent within the dark chamber between the initial and final preference tests was used to calculate a CPA score. Aversion increased with increasing isoflurane concentration: from 1.7% to 2.7% to 3.7% isoflurane, mean ± SE CPA score decreased from 19.6 ± 20.1 s to -25.6 ± 23.2 s, to -116.9 ± 30.6 s (F1,54 = 15.4, p < 0.001). Our results suggest that, when using the drop method to administer isoflurane, concentrations between 1.7% and 2.7% can be used to minimize female mouse aversion to induction.

许多指南都认为二氧化碳安乐死前的异氟烷麻醉是一种改进。没有蒸发器的机构可以使用 "滴注 "法,即将液态麻醉剂引入诱导室。了解异氟醚的最小厌恶浓度至关重要。之前的研究表明,使用滴注法给小鼠注射浓度为 5%的异氟醚会使其产生厌恶感。其他工作表明,较低浓度(1.7% 至 3.7%)的异氟烷也可用于滴注法麻醉小鼠,但对这些浓度的厌恶程度尚未进行测试。我们使用条件性场所厌恶(CPA)范式评估了小鼠对这些较低浓度异氟醚滴注法的厌恶程度。雌性 C57BL/6J (OT-1) 小鼠(n = 28)被随机分配到三种异氟醚浓度中的一种:1.7%、2.7% 和 3.7%。小鼠适应光-暗装置。在进行暗室(+异氟醚)和光室调节之前和之后,小鼠接受了初始和最终偏好评估;初始和最终偏好测试之间小鼠在暗室内停留时间的变化被用来计算 CPA 分数。厌恶程度随着异氟烷浓度的增加而增加:异氟烷浓度从 1.7% 到 2.7% 再到 3.7%,CPA 评分的平均值(±SE)从 19.6 ± 20.1 秒下降到 -25.6 ± 23.2 秒,再下降到 -116.9 ± 30.6 秒(F1,54 = 15.4,p.
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引用次数: 0
How cage effects can hurt statistical analyses of completely randomized designs. 笼效应如何影响完全随机设计的统计分析?
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-01 Epub Date: 2024-09-24 DOI: 10.1177/00236772241276785
Reid D Landes

Cage effects: some researchers worry about them, some don't, and some aren't even aware of them. When statistical analyses do not account for cage effects, there is real reason to worry. Regardless of researchers' worries or lack thereof, all researchers should be aware of how cage effects can affect the results. The "how" depends, in part, on the experimental design. Here, I (a) define cage effects; (b) illustrate a completely randomized design (CRD) often used in animal experiments; (c) explain how statistical significance is artificially inflated when cage effects are ignored and (d) give guidance on proper analyses and on how to increase statistical power in CRDs.

笼子效应:有些研究人员担心笼子效应,有些不担心,有些甚至没有意识到笼子效应。当统计分析没有考虑笼子效应时,确实有理由担心。无论研究人员是否担心,所有研究人员都应该意识到笼效应会如何影响研究结果。至于 "如何",部分取决于实验设计。在此,我将(a) 给笼子效应下一个定义;(b) 举例说明动物实验中经常使用的完全随机设计(CRD);(c) 解释当笼子效应被忽视时,统计显著性是如何被人为夸大的;(d) 指导如何进行正确的分析,以及如何提高 CRD 的统计功率。
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引用次数: 0
Methods for applying blinding and randomisation in animal experiments. 在动物实验中应用盲法和随机化的方法。
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-01 DOI: 10.1177/00236772241272991
P S Verhave, R van Eenige, Iacw Tiebosch

Blinding and randomisation are important methods for increasing the robustness of pre-clinical studies, as incomplete or improper implementation thereof is recognised as a source of bias. Randomisation ensures that any known and unknown covariates introducing bias are randomly distributed over the experimental groups. Thereby, differences between the experimental groups that might otherwise have contributed to false positive or -negative results are diminished. Methods for randomisation range from simple randomisation (e.g. rolling a dice) to advanced randomisation strategies involving the use of specialised software. Blinding on the other hand ensures that researchers are unaware of group allocation during the preparation, execution and acquisition and/or the analysis of the data. This minimises the risk of unintentional influences resulting in bias. Methods for blinding require strong protocols and a team approach. In this review, we outline methods for randomisation and blinding and give practical tips on how to implement them, with a focus on animal studies.

盲法和随机化是提高临床前研究稳健性的重要方法,因为盲法和随机化的不完全或不当实施被认为是偏差的来源。随机化可确保任何会产生偏差的已知和未知协变量在实验组中随机分布。这样,实验组之间的差异就会减少,否则可能会造成假阳性或假阴性结果。随机化的方法多种多样,从简单的随机化(如掷骰子)到使用专门软件的高级随机化策略,不一而足。另一方面,盲法可确保研究人员在准备、执行、获取和/或分析数据期间不知道组别分配情况。这就最大限度地降低了无意影响导致偏差的风险。盲法需要强有力的协议和团队合作。在这篇综述中,我们概述了随机化和盲法的方法,并以动物研究为重点,提供了如何实施这些方法的实用技巧。
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引用次数: 0
Study design: think 'scientific value' not 'p-values'. 研究设计:要考虑 "科学价值",而不是 "P 值"。
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-01 DOI: 10.1177/00236772241276806
Penny S Reynolds

Statistically based experimental designs have been available for over a century. However, many preclinical researchers are completely unaware of these methods, and the success of experiments is usually equated only with 'p < 0.05'. By contrast, a well-thought-out experimental design strategy provides data with evidentiary and scientific value. A value-based strategy requires implementation of statistical design principles coupled with basic project management techniques. This article outlines the three phases of a value-based design strategy: proper framing of the research question, statistically based operationalisation through careful selection and structuring of appropriate inputs, and incorporation of methods that minimise bias and process variation. Appropriate study design increases study validity and the evidentiary strength of the results, reduces animal numbers, and reduces waste from noninformative experiments. Statistically based experimental design is thus a key component of the 'Reduction' pillar of the 3R (Replacement, Reduction, Refinement) principles for ethical animal research.

以统计学为基础的实验设计已有一个多世纪的历史。然而,许多临床前研究人员完全不了解这些方法,而且实验的成功与否通常只等同于 "p"。
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引用次数: 0
Depicting variability and uncertainty using intervals and error bars. 使用区间和误差条描述可变性和不确定性。
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-01 Epub Date: 2024-09-05 DOI: 10.1177/00236772241247105
Naomi Altman, Martin Krzywinski

Variability is inherent in most biological systems due to differences among members of the population. Two types of variation are commonly observed in studies: differences among samples and the "error" in estimating a population parameter (e.g. mean) from a sample. While these concepts are fundamentally very different, the associated variation is often expressed using similar notation-an interval that represents a range of values with a lower and upper bound. In this article we discuss how common intervals are used (and misused).

由于种群成员之间的差异,大多数生物系统都存在固有的变异性。研究中通常会观察到两种类型的变异:样本间的差异和根据样本估计群体参数(如平均值)时的 "误差"。虽然这两个概念从根本上说非常不同,但相关的变异通常使用类似的符号来表示--一个区间,代表一个有下限和上限的数值范围。本文将讨论常见区间的使用(和误用)方式。
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引用次数: 0
About statistical significance, and the lack thereof. 关于统计意义,以及缺乏统计意义的问题。
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-01 Epub Date: 2024-08-19 DOI: 10.1177/00236772241248509
Fulvio Magara, Benjamin Boury-Jamot

Absence of statistical significance (i.e., p > 0.05) in the results of a frequentist test comparing two samples is often used as evidence of absence of difference, or absence of effect of a treatment, on the measured variable. Such conclusions are often wrong because absence of significance may merely result from a sample size that is too small to reveal an effect. To conclude that there is no meaningful effect of a treatment/condition, it is necessary to use an appropriate statistical approach. For frequentist statistics, a simple tool for this goal is the 'two one-sided t-test,' a form of equivalence test that relies on the a priori definition of a minimal difference considered to be relevant. In other words, the smallest effect size of interest should be established in advance. We present the principles of this test and give examples where it allows correct interpretation of the results of a classical t-test assuming absence of difference. Equivalence tests are also very useful in probing whether certain significant results are also biologically meaningful, because when comparing large samples it is possible to find significant results in both an equivalence test and in a two-sample t-test, assuming no difference as the null hypothesis.

在比较两个样本的频数检验结果中,如果没有统计学意义(即 p > 0.05),通常会被用来证明在测量的变量上没有差异,或治疗没有效果。这种结论往往是错误的,因为没有显著性可能只是因为样本量太小,无法显示效果。要得出某项治疗/条件不存在有意义影响的结论,必须使用适当的统计方法。对于频数统计来说,实现这一目标的简单工具是 "两个单侧 t 检验",这是一种等效检验,它依赖于被认为相关的最小差异的先验定义。换句话说,应事先确定相关的最小效应大小。我们介绍了这种检验的原理,并举例说明了在假设无差异的情况下,它可以正确解释经典 t 检验的结果。等效检验在探究某些显著结果是否也具有生物学意义方面也非常有用,因为在比较大样本时,假设无差异为零假设,在等效检验和双样本 t 检验中都有可能发现显著结果。
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引用次数: 0
Testing for normality: a user's (cautionary) guide. 正态性测试:用户(警示)指南。
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-01 DOI: 10.1177/00236772241276808
Romain-Daniel Gosselin

The normality assumption postulates that empirical data derives from a normal (Gaussian) population. It is a pillar of inferential statistics that enables the theorization of probability functions and the computation of p-values thereof. The breach of this assumption may not impose a formal mathematical constraint on the computation of inferential outputs (e.g., p-values) but may make them inoperable and possibly lead to unethical waste of laboratory animals. Various methods, including statistical tests and qualitative visual examination, can reveal incompatibility with normality and the choice of a procedure should not be trivialized. The following minireview will provide a brief overview of diagrammatical methods and statistical tests commonly employed to evaluate congruence with normality. Special attention will be given to the potential pitfalls associated with their application. Normality is an unachievable ideal that practically never accurately describes natural variables, and detrimental consequences of non-normality may be safeguarded by using large samples. Therefore, the very concept of preliminary normality testing is also, arguably provocatively, questioned.

正态性假设假定经验数据来自正常(高斯)群体。它是推理统计的支柱,使概率函数的理论化及其 p 值的计算成为可能。违反这一假设可能不会对推理输出(如 p 值)的计算造成正式的数学限制,但可能会使其无法操作,并可能导致实验动物的不道德浪费。各种方法,包括统计检验和定性直观检查,都可以发现不符合正态性的情况,因此不应轻视程序的选择。下面的小视图将简要介绍常用于评估是否符合正态性的图解法和统计检验法。我们将特别关注与这些方法的应用相关的潜在隐患。正态性是一个无法实现的理想,实际上永远无法准确描述自然变量,而使用大样本可以避免非正态性的有害后果。因此,初步正态性检验的概念本身也受到了质疑,可以说是挑衅性的质疑。
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引用次数: 0
Ditching the norm: Using alternative distributions for biological data analysis. 抛弃常规:使用替代分布进行生物数据分析
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-01 Epub Date: 2024-08-19 DOI: 10.1177/00236772241246602
Stanley E Lazic

Most classical statistical tests assume data are normally distributed. If this assumption is not met, researchers often turn to non-parametric methods. These methods have some drawbacks, and if no suitable non-parametric test exists, a normal distribution may be used inappropriately instead. A better option is to select a distribution appropriate for the data from dozens available in modern software packages. Selecting a distribution that represents the data generating process is a crucial but overlooked step in analysing data. This paper discusses several alternative distributions and the types of data that they are suitable for.

大多数经典统计检验都假设数据呈正态分布。如果不符合这一假设,研究人员通常会求助于非参数方法。这些方法有一些缺点,如果没有合适的非参数检验方法,可能会不恰当地使用正态分布。更好的选择是从现代软件包中的几十种分布中选择适合数据的分布。选择一个能代表数据生成过程的分布是分析数据的关键步骤,但却被忽视了。本文将讨论几种可供选择的分布及其适用的数据类型。
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引用次数: 0
Understanding p-values and significance. 了解 p 值和显著性。
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-01 Epub Date: 2024-09-24 DOI: 10.1177/00236772241247106
Naomi Altman, Martin Krzywinski

P-values combined with estimates of effect size are used to assess the importance of experimental results. However, their interpretation can be invalidated by selection bias when testing multiple hypotheses, fitting multiple models or even informally selecting results that seem interesting after observing the data. We offer an introduction to principled uses of p-values (targeted at the non-specialist) and identify questionable practices to be avoided.

P 值与效应大小估计值相结合,用于评估实验结果的重要性。然而,在测试多个假设、拟合多个模型或甚至在观察数据后非正式地选择看起来有趣的结果时,其解释可能会因选择偏差而失效。我们将介绍 p 值的原则性用法(针对非专业人士),并指出应避免的可疑做法。
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引用次数: 0
Laboratory Animals presents Special Issue Biostatistics Notes. 实验动物》推出特刊《生物统计笔记》。
IF 1.3 4区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2024-10-01 DOI: 10.1177/00236772241273059
Romain-Daniel Gosselin, Penny S Reynolds
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引用次数: 0
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Laboratory Animals
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