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An automatic classification method of testicular histopathology based on SC-YOLO framework. 基于 SC-YOLO 框架的睾丸组织病理学自动分类方法。
IF 2.7 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-12 DOI: 10.1080/07366205.2024.2393544
Jinggen Wu,Yao Sun,Yangbo Jiang,Yangcheng Bu,Chong Chen,Jingping Li,Lejun Li,Weikang Chen,Keren Cheng,Jian Xu
The pathological diagnosis and treatment of azoospermia depend on precise identification of spermatogenic cells. Traditional methods are time-consuming and highly subjective due to complexity of Johnsen score, posing challenges for accurately diagnosing azoospermia. Here, we introduce a novel SC-YOLO framework for automating the classification of spermatogenic cells that integrates S3Ghost module, CoordAtt module and DCNv2 module, effectively capturing texture and shape features of spermatogenic cells while reducing model parameters. Furthermore, we propose a simplified Johnsen score criteria to expedite the diagnostic process. Our SC-YOLO framework presents the higher efficiency and accuracy of deep learning technology in spermatogenic cell recognition. Future research endeavors will focus on optimizing the model's performance and exploring its potential for clinical applications.
无精子症的病理诊断和治疗取决于对生精细胞的精确鉴定。传统方法耗时长,而且由于约翰森评分的复杂性,主观性很强,给无精子症的准确诊断带来了挑战。在此,我们介绍了一种用于自动分类生精细胞的新型 SC-YOLO 框架,该框架集成了 S3Ghost 模块、CoordAtt 模块和 DCNv2 模块,可有效捕捉生精细胞的纹理和形状特征,同时减少模型参数。此外,我们还提出了简化的约翰森评分标准,以加快诊断过程。我们的 SC-YOLO 框架展示了深度学习技术在生精细胞识别中的更高效率和准确性。未来的研究工作将侧重于优化模型的性能,并探索其在临床应用中的潜力。
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引用次数: 0
When is an SNP not an SNP? 什么时候 SNP 不是 SNP?
IF 2.7 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-12 DOI: 10.1080/07366205.2024.2387993
Shapour Jalilzadeh,Valerie Walker,Gary P Leggatt,Eli Hatchwell
Genomic duplications are important sources of structural change and gene innovation. In humans, the most recent and highly identical sequences (>90% homology, >1 kb long) are known as segmental duplications (SDs). Single-nucleotide variants or single-nucleotide polymorphisms within SDs have not been systematically assessed due to limitations around mapping short-read sequencing data. Single-nucleotide variant rs62486260 was flagged in a study of familial renal stone disease but it was unclear whether it was real or an artifact resulting from the presence of a SD. We describe in silico and wet-lab approaches to investigate this, using segment-specific long-PCR assays, followed by short PCR for Sanger sequencing. Our conclusion was that rs62486260 is an artifact. Our approach can be generalized to deal with other such situations.
基因组复制是结构变化和基因创新的重要来源。在人类中,最新和高度相同的序列(同源性大于 90%,长度大于 1 kb)被称为片段重复(SD)。由于短线程测序数据映射的局限性,SDs 内的单核苷酸变异或单核苷酸多态性尚未得到系统评估。在一项关于家族性肾结石病的研究中,单核苷酸变异体 rs62486260 被标记出来,但尚不清楚它是真实的还是因 SD 的存在而产生的假象。我们介绍了利用分段特异性长 PCR 检测和用于 Sanger 测序的短 PCR 进行研究的硅学和湿实验室方法。我们的结论是,rs62486260 是一个假象。我们的方法可用于处理其他此类情况。
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引用次数: 0
Sampling and analysis methods of air-borne microorganisms in hospital air: a review. 医院空气中气载微生物的采样和分析方法:综述。
IF 2.7 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-12 DOI: 10.1080/07366205.2024.2372939
Xinyan Wei,Xuezheng Ma,Feng Tian,Zhaohui Wei,Liping Zhang,Kongxin Hu
Pathogenic microorganisms can spread in the air as bioaerosols. When the human body is exposed to different bioaerosols, various infectious diseases may occur. As indoor diagnosis and treatment environments, hospitals are relatively closed and have a large flow rate of people. This indoor environment contains complex aerosol components; therefore, effective sampling and detection of microbial elements are essential in airborne pathogen monitoring. This article reviews the sampling and detection of different kinds of microorganisms in bioaerosols from indoor diagnostic and therapeutic settings, with a particular focus on microbial activity. This provides deeper insights into bioaerosols in diagnostic and therapeutic settings.
病原微生物会以生物气溶胶的形式在空气中传播。当人体接触到不同的生物气溶胶时,可能会引发各种传染病。医院作为室内诊疗环境,相对封闭,人流量大。这种室内环境含有复杂的气溶胶成分,因此,有效采样和检测微生物元素对空气中病原体的监测至关重要。本文回顾了对室内诊断和治疗环境生物气溶胶中不同种类微生物的采样和检测,尤其关注微生物的活性。这有助于深入了解诊断和治疗环境中的生物气溶胶。
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引用次数: 0
Mito-kaede photoactivation and chase experiment for mitophagy: optimizing flux measurement via fluid exchange system. 有丝分裂的光激活和追逐实验:通过流体交换系统优化通量测量。
IF 2.7 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-11 DOI: 10.1080/07366205.2024.2372955
Hiroyuki Morinaga,Yoh Sugawara,Yoshinori Kitagawa,Jingyuan Chen,Nobuo Yasuda,Hiroki Ogata,Yoshihiro Yamaguchi,Masao Kaneki,Joseph A Jeevendra Martyn,Shingo Yasuhara
Modulating autophagy and mitophagy, vital cellular quality control systems, offer therapeutic potential for critical illnesses. However, limited drug screening options hinder progress. We present a novel assay using the photoswitchable fluorescent reporter, mito-Kaede, to quantify mitophagy flux. Mito-Kaede's superior UV-induced photoconversion and brightness post-conversion make it ideal for prolonged mitochondrial dynamics tracking. Its specificity in responding to mitophagy, confirmed by parkin-knockout cells, adds value. When coupled with a custom fluid exchange system, enabling efficient medium changes, precise mitophagy observations become feasible. This mitophagy assay, alongside our methodological insights, can decipher mitophagy's role in pathology and supports drug screening efforts.
自噬和有丝分裂是重要的细胞质量控制系统,调节这两个系统可为危重疾病提供治疗潜力。然而,有限的药物筛选方案阻碍了研究的进展。我们介绍了一种利用光开关荧光报告物 mito-Kaede 来量化有丝分裂通量的新型检测方法。Mito-Kaede 具有优异的紫外线诱导光电转换能力和转换后的亮度,因此非常适合用于长时间的线粒体动态跟踪。经 Parkin 基因敲除细胞证实,它对有丝分裂反应的特异性增加了其价值。如果与定制的液体交换系统配合使用,就能有效地更换培养基,从而实现对有丝分裂的精确观察。这种有丝分裂检测方法与我们的方法论见解相结合,可以破译有丝分裂在病理学中的作用,并为药物筛选工作提供支持。
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引用次数: 0
Prioritizing privacy and presentation of supportable hypothesis testing in forensic genetic genealogy investigations. 在法医遗传系谱调查中优先考虑隐私和提出可支持的假设检验。
IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-09 DOI: 10.1080/07366205.2024.2386218
Bruce Budowle, Lee Baker, Antti Sajantila, Kristen Mittelman, David Mittelman

Investigative leads are not generated by traditional forensic DNA testing, if the source of the forensic evidence or a 1st degree relative of unidentified human remains is not in the DNA database. In such cases, forensic genetic genealogy (FGG) can provide valuable leads. However, FGG generated genetic data contain private and sensitive information. Therefore, it is essential to deploy approaches that minimize unnecessary disclosure of these data to mitigate potential risks to individual privacy. We recommend protective practices that need not impact effective reporting of relationship identifications. Examples include performing one-to-one comparisons of DNA profiles of third-party samples and evidence samples offline with an "air gap" to the internet and shielding the specific shared single nucleotide polymorphisms (SNP) states and locations by binning adjacent SNPs in forensic reports. Such approaches reduce risk of unwanted access to or reverse engineering of third-party individuals' genetic data and can give these donors greater confidence to support use of their DNA profiles in FGG investigation.

如果法医证据的来源或身份不明遗骸的一级亲属不在 DNA 数据库中,传统的法医 DNA 检测就无法产生调查线索。在这种情况下,法医基因谱(FGG)可以提供有价值的线索。然而,FGG 生成的基因数据包含隐私和敏感信息。因此,必须采取尽量减少不必要披露这些数据的方法,以降低对个人隐私的潜在风险。我们建议采取不影响有效报告关系识别的保护措施。例如,对第三方样本和证据样本的脱机 DNA 图谱进行一对一的比较,并与互联网保持 "间隙";在法医报告中对相邻 SNP 进行分选,以屏蔽特定的共享单核苷酸多态性 (SNP) 状态和位置。这种方法降低了意外访问或逆向工程第三方个人基因数据的风险,并可使这些捐献者更有信心支持在 FGG 调查中使用其 DNA 图谱。
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引用次数: 0
Dual-mode detection of glucose based on pistol-like DNAzyme-mediated exonuclease-assisted signal cycle. 基于手枪式 DNA 酶介导的外切酶辅助信号循环的葡萄糖双模式检测。
IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-05 DOI: 10.1080/07366205.2024.2381403
Huiying Lu, Xiaofeng Wang

Detecting glucose accurately and sensitively from clinical samples like tears and saliva is still difficult. We have created a sensor that can detect glucose with high sensitivity and accuracy by combining the use of glucose oxidase (GOx) to catalyze glucose, a pistol-like DNAzyme (PLDz) to transform the signal, gold nanoparticles (AuNPs) to enhance the optical properties and the exonuclease-III (Exo-III) to amplify the signal. As a result, the proposed method exhibits a low detection limit of 7.5 pM and a wide detection range covering seven orders of magnitude. The suggested dual-mode strategy provides a sensitive, precise and specific detection method for glucose. Another advantage is that the dual-mode technique significantly improves the precision and consistency of the measurements, demonstrating its immense potential for use in biomedical research and clinical diagnostics.

要从泪液和唾液等临床样本中准确而灵敏地检测葡萄糖仍然很困难。我们利用葡萄糖氧化酶(GOx)催化葡萄糖、手枪样 DNA 酶(PLDz)转换信号、金纳米粒子(AuNPs)增强光学特性以及外切酶-III(Exo-III)放大信号,创造出了一种能够高灵敏度、高准确度检测葡萄糖的传感器。因此,该方法的检测限低至 7.5 pM,检测范围广达七个数量级。建议的双模式策略提供了一种灵敏、精确和特异的葡萄糖检测方法。另一个优点是,双模式技术大大提高了测量的精确度和一致性,显示了其在生物医学研究和临床诊断中的巨大应用潜力。
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引用次数: 0
The environmental impact of AI in the lab: a double-edged sword? 实验室人工智能对环境的影响:一把双刃剑?
IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-31 DOI: 10.1080/07366205.2024.2376459
Annie Coulson

Computational tools, particularly AI, are becoming more ubiquitous in scientific research; but what impact do they have on the environment?[Formula: see text].

计算工具,尤其是人工智能,在科学研究中越来越无处不在;但它们对环境有什么影响?
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引用次数: 0
Modular probe-based colorimetric miRNA detection via polymerase/endonuclease assisted chain displacement. 通过聚合酶/内切酶辅助链置换进行基于探针的模块化比色 miRNA 检测。
IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-23 DOI: 10.1080/07366205.2024.2368394
Xialing Xu, Ping Zhang, Siyu Tao

Methods for sequence-specific microRNA (miRNA) analysis are crucial for miRNA research and guiding nursing strategies. We have devised a colorimetric technique for detecting miRNA using a dumbbell probe-based polymerase/endonuclease assisted chain displacement, along with silver ions (Ag+) aptamer assisted color reaction. The suggested approach enables precise measurement of miRNA-21 within the concentration range of 100 fM-5 nM, with a low detection limit of 45.32 fM. Additionally, it exhibits exceptional capability in distinguishing variations at the level of individual nucleotides. Furthermore, the detection technique may be utilized to precisely measure the amount of miRNA-21 in serum samples, demonstrating a high level of concordance with the findings obtained from a commercially available miRNA detection kit.

序列特异性 microRNA(miRNA)分析方法对于 miRNA 研究和指导护理策略至关重要。我们设计了一种利用哑铃探针聚合酶/内切酶辅助链转移以及银离子(Ag+)合剂辅助显色反应来检测 miRNA 的显色技术。所建议的方法能在 100 fM-5 nM 的浓度范围内精确测量 miRNA-21,检测限低至 45.32 fM。此外,它在区分单个核苷酸水平的变化方面表现出卓越的能力。此外,该检测技术还可用于精确测量血清样本中 miRNA-21 的含量,与市售 miRNA 检测试剂盒的结果高度一致。
{"title":"Modular probe-based colorimetric miRNA detection <i>via</i> polymerase/endonuclease assisted chain displacement.","authors":"Xialing Xu, Ping Zhang, Siyu Tao","doi":"10.1080/07366205.2024.2368394","DOIUrl":"https://doi.org/10.1080/07366205.2024.2368394","url":null,"abstract":"<p><p>Methods for sequence-specific microRNA (miRNA) analysis are crucial for miRNA research and guiding nursing strategies. We have devised a colorimetric technique for detecting miRNA using a dumbbell probe-based polymerase/endonuclease assisted chain displacement, along with silver ions (Ag<sup>+</sup>) aptamer assisted color reaction. The suggested approach enables precise measurement of miRNA-21 within the concentration range of 100 fM-5 nM, with a low detection limit of 45.32 fM. Additionally, it exhibits exceptional capability in distinguishing variations at the level of individual nucleotides. Furthermore, the detection technique may be utilized to precisely measure the amount of miRNA-21 in serum samples, demonstrating a high level of concordance with the findings obtained from a commercially available miRNA detection kit.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141747399","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Adipocyte ABCA1 expression analysis using flow cytometry. 使用流式细胞仪分析脂肪细胞 ABCA1 的表达。
IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-17 DOI: 10.1080/07366205.2024.2376466
Sakshi Shukla, Ashutosh Bansal, Sandeep Aggarwal, Archna Singh

Adipocyte characterization and assessing membrane proteins using flow cytometry has been proven to be challenging as adipocytes are fragile, especially in subjects with high BMI. We overcame these challenges through a protocol optimizing tissue digestion time by reducing intermediate steps to minimize adipocyte friction and breakage. We avoided requirement for specialized instrument configuration and used a modified gating strategy to prevent inclusion of lipid droplets during analysis. Up to 90% of the cell population were available in the gating area. We checked the expression level of ABCA1, a membrane protein reaffirming adipocyte selection. In summary, this protocol requires lesser tissue sample improving feasibility and cost efficiency. Thus, our flow cytometry method is an improvement for studying adipocyte membrane characteristics.

使用流式细胞术鉴定脂肪细胞特征和评估膜蛋白已被证明具有挑战性,因为脂肪细胞很脆弱,尤其是在高体重指数的受试者中。我们通过减少中间步骤来优化组织消化时间,最大程度地减少脂肪细胞的摩擦和破损,从而克服了这些挑战。我们避免了对专业仪器配置的要求,并采用了改进的门控策略,以防止在分析过程中加入脂滴。多达 90% 的细胞群可进入选通区域。我们检测了 ABCA1 的表达水平,这是一种膜蛋白,再次证实了脂肪细胞的选择。总之,该方案所需的组织样本较少,提高了可行性和成本效益。因此,我们的流式细胞仪方法是研究脂肪细胞膜特征的一种改进。
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引用次数: 0
Novel buffer for long-term preservation of DNA in biological material at room temperature1. 用于在室温下长期保存生物材料中 DNA 的新型缓冲液。
IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-01 DOI: 10.1080/07366205.2024.2360813
Mohaimin Kasu, Peter G Ristow, Adria Michelle Burrows, Zafrir Kuplik, Mark J Gibbons, Maria E D'Amato

The collection and preservation of biological material before DNA analysis is critical for inter alia biomedical research, medical diagnostics, forensics and biodiversity conservation. In this study, we evaluate an in-house formulated buffer called the Forensic DNA Laboratory-buffer (FDL-buffer) for preservation of biological material for long term at room temperature. Human saliva stored in the buffer for 8 years, human blood stored for 3 years and delicate animal tissues from the jellyfish Pelagia noctiluca comb jelly Beroe sp., stored for 4 and 6 years respectively consistently produced high-quality DNA. FDL-buffer exhibited compatibility with standard organic, salting out and spin-column extraction methods, making it versatile and applicable to a wide range of applications, including automation.

在 DNA 分析前收集和保存生物材料对于生物医学研究、医疗诊断、法医和生物多样性保护等至关重要。在这项研究中,我们对一种内部配制的缓冲液进行了评估,这种缓冲液被称为法医 DNA 实验室缓冲液(FDL-buffer),用于在室温下长期保存生物材料。在缓冲液中保存 8 年的人类唾液、保存 3 年的人类血液以及分别保存 4 年和 6 年的海蜇(Pelagia noctiluca comb jelly Beroe sp.FDL 缓冲液与标准的有机提取、盐析提取和旋柱提取方法兼容,因此它用途广泛,适用于包括自动化在内的各种应用。
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引用次数: 0
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BioTechniques
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