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  1. Article ; Online: Improving the quality of quantitative polymerase chain reaction experiments: 15 years of MIQE.

    Bustin, Stephen A

    Molecular aspects of medicine

    2024  Volume 96, Page(s) 101249

    Abstract: The quantitative polymerase chain reaction (qPCR) is fundamental to molecular biology. It is not just a laboratory technique, qPCR is a bridge between research and clinical practice. Its theoretical foundations guide the design of experiments, while its ... ...

    Abstract The quantitative polymerase chain reaction (qPCR) is fundamental to molecular biology. It is not just a laboratory technique, qPCR is a bridge between research and clinical practice. Its theoretical foundations guide the design of experiments, while its practical implications extend to diagnostics, treatment, and research advancements in the life sciences, human and veterinary medicine, agriculture, and forensics. However, the accuracy, reliability and reproducibility of qPCR data face challenges arising from various factors associated with experimental design, execution, data analysis and inadequate reporting details. Addressing these concerns, the Minimum Information for the Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines have emerged as a cohesive framework offering a standardised set of recommendations that describe the essential information required for assessing qPCR experiments. By emphasising the importance of methodological rigour, the MIQE guidelines have made a major contribution to improving the trustworthiness, consistency, and transparency of many published qPCR results. However, major challenges related to awareness, resources, and publication pressures continue to affect their consistent application.
    MeSH term(s) Humans ; Reproducibility of Results ; Real-Time Polymerase Chain Reaction/methods
    Language English
    Publishing date 2024-01-29
    Publishing country England
    Document type Journal Article ; Review
    ZDB-ID 197640-0
    ISSN 1872-9452 ; 0098-2997
    ISSN (online) 1872-9452
    ISSN 0098-2997
    DOI 10.1016/j.mam.2024.101249
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

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  2. Article ; Online: Molecular Pathology, Diagnostics and Therapeutics: A Story of Success in 2022.

    Bustin, Stephen A

    International journal of molecular sciences

    2023  Volume 24, Issue 5

    Abstract: Molecular pathology, diagnostics and therapeutics are three closely related topics of critical importance in medical research and clinical practice [ ... ]. ...

    Abstract Molecular pathology, diagnostics and therapeutics are three closely related topics of critical importance in medical research and clinical practice [...].
    MeSH term(s) Pathology, Molecular
    Language English
    Publishing date 2023-03-06
    Publishing country Switzerland
    Document type Editorial
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms24055063
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  3. Article ; Online: Advances in Molecular Medicine: Unravelling Disease Complexity and Pioneering Precision Healthcare.

    Bustin, Stephen A / Jellinger, Kurt A

    International journal of molecular sciences

    2023  Volume 24, Issue 18

    Abstract: The escalating impacts of the climate crisis, zoonotic spill-over, and antibiotic resistance have positioned molecular medicine at the forefront of pioneering translational research [ ... ]. ...

    Abstract The escalating impacts of the climate crisis, zoonotic spill-over, and antibiotic resistance have positioned molecular medicine at the forefront of pioneering translational research [...].
    Language English
    Publishing date 2023-09-16
    Publishing country Switzerland
    Document type Editorial
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms241814168
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  4. Article ; Online: Molecular Pathology, Diagnostics and Therapeutics

    Stephen A. Bustin

    International Journal of Molecular Sciences, Vol 24, Iss 5063, p

    A Story of Success in 2022

    2023  Volume 5063

    Abstract: Molecular pathology, diagnostics and therapeutics are three closely related topics of critical importance in medical research and clinical practice [.] ...

    Abstract Molecular pathology, diagnostics and therapeutics are three closely related topics of critical importance in medical research and clinical practice [.]
    Keywords n/a ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
    Language English
    Publishing date 2023-03-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: FlashPCR: Revolutionising qPCR by Accelerating Amplification through Low ∆T Protocols.

    Bustin, Stephen A / Kirvell, Sara / Nolan, Tania / Shipley, Gregory L

    International journal of molecular sciences

    2024  Volume 25, Issue 5

    Abstract: Versatility, sensitivity, and accuracy have made the real-time polymerase chain reaction (qPCR) a crucial tool for research, as well as diagnostic applications. However, for point-of-care (PoC) use, traditional qPCR faces two main challenges: long run ... ...

    Abstract Versatility, sensitivity, and accuracy have made the real-time polymerase chain reaction (qPCR) a crucial tool for research, as well as diagnostic applications. However, for point-of-care (PoC) use, traditional qPCR faces two main challenges: long run times mean results are not available for half an hour or more, and the requisite high-temperature denaturation requires more robust and power-demanding instrumentation. This study addresses both issues and revises primer and probe designs, modified buffers, and low ∆T protocols which, together, speed up qPCR on conventional qPCR instruments and will allow for the development of robust, point-of-care devices. Our approach, called "FlashPCR", uses a protocol involving a 15-second denaturation at 79 °C, followed by repeated cycling for 1 s at 79 °C and 71 °C, together with high Tm primers and specific but simple buffers. It also allows for efficient reverse transcription as part of a one-step RT-qPCR protocol, making it universally applicable for both rapid research and diagnostic applications.
    MeSH term(s) Real-Time Polymerase Chain Reaction/methods ; Reverse Transcription ; Sensitivity and Specificity
    Language English
    Publishing date 2024-02-28
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms25052773
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Advances in Molecular Medicine

    Stephen A. Bustin / Kurt A. Jellinger

    International Journal of Molecular Sciences, Vol 24, Iss 14168, p

    Unravelling Disease Complexity and Pioneering Precision Healthcare

    2023  Volume 14168

    Abstract: The escalating impacts of the climate crisis, zoonotic spill-over, and antibiotic resistance have positioned molecular medicine at the forefront of pioneering translational research [.] ...

    Abstract The escalating impacts of the climate crisis, zoonotic spill-over, and antibiotic resistance have positioned molecular medicine at the forefront of pioneering translational research [.]
    Keywords n/a ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
    Language English
    Publishing date 2023-09-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

    Full text online

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    Inter-library loan at ZB MED

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  7. Article: How to speed up the polymerase chain reaction.

    Bustin, Stephen A

    Biomolecular detection and quantification

    2017  Volume 12, Page(s) 10–14

    Abstract: Reducing the time taken to run qPCR assays on today's qPCR cyclers is rather straightforward and requires no specialised reagents or instruments. As the first article in a new series of short technical reports, I demonstrate that it is possible to reduce ...

    Abstract Reducing the time taken to run qPCR assays on today's qPCR cyclers is rather straightforward and requires no specialised reagents or instruments. As the first article in a new series of short technical reports, I demonstrate that it is possible to reduce significantly both denaturation temperatures and cycling times, whilst retaining sensitivity and specificity of the original qPCR conditions.
    Language English
    Publishing date 2017-06-20
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 2821770-6
    ISSN 2214-7535
    ISSN 2214-7535
    DOI 10.1016/j.bdq.2017.05.002
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: RT-qPCR Testing of SARS-CoV-2: A Primer.

    Bustin, Stephen A / Nolan, Tania

    International journal of molecular sciences

    2020  Volume 21, Issue 8

    Abstract: Testing for the presence of coronavirus is an essential diagnostic tool for monitoring and managing the current COVID-19 pandemic. The only reliable test in current use for testing acute infection targets the genome of SARS-CoV-2, and the most widely ... ...

    Abstract Testing for the presence of coronavirus is an essential diagnostic tool for monitoring and managing the current COVID-19 pandemic. The only reliable test in current use for testing acute infection targets the genome of SARS-CoV-2, and the most widely used method is quantitative fluorescence-based reverse transcription polymerase chain reaction (RT-qPCR). Despite its ubiquity, there is a significant amount of uncertainty about how this test works, potential throughput and reliability. This has resulted in widespread misrepresentation of the problems faced using this test during the current COVID-19 epidemic. This primer provides simple, straightforward and impartial information about RT-qPCR.
    MeSH term(s) Betacoronavirus/genetics ; Betacoronavirus/isolation & purification ; COVID-19 ; COVID-19 Testing ; Clinical Laboratory Techniques/methods ; Coronavirus Infections/diagnosis ; Coronavirus Infections/virology ; DNA Primers ; Pandemics ; Pneumonia, Viral/diagnosis ; Pneumonia, Viral/virology ; RNA, Viral/genetics ; RNA, Viral/isolation & purification ; Real-Time Polymerase Chain Reaction/methods ; SARS-CoV-2
    Chemical Substances DNA Primers ; RNA, Viral
    Keywords covid19
    Language English
    Publishing date 2020-04-24
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms21083004
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Temporal gene signature of myofibroblast transformation in Peyronie's disease: first insights into the molecular mechanisms of irreversibility.

    Ilg, Marcus M / Harding, Sophie / Lapthorn, Alice R / Kirvell, Sara / Ralph, David J / Bustin, Stephen A / Ball, Graham / Cellek, Selim

    The journal of sexual medicine

    2024  Volume 21, Issue 4, Page(s) 278–287

    Abstract: Background: Transformation of resident fibroblasts to profibrotic myofibroblasts in the tunica albuginea is a critical step in the pathophysiology of Peyronie's disease (PD). We have previously shown that myofibroblasts do not revert to the fibroblast ... ...

    Abstract Background: Transformation of resident fibroblasts to profibrotic myofibroblasts in the tunica albuginea is a critical step in the pathophysiology of Peyronie's disease (PD). We have previously shown that myofibroblasts do not revert to the fibroblast phenotype and we suggested that there is a point of no return at 36 hours after induction of the transformation. However, the molecular mechanisms that drive this proposed irreversibility are not known.
    Aim: Identify molecular pathways that drive the irreversibility of myofibroblast transformation by analyzing the expression of the genes involved in the process in a temporal fashion.
    Methods: Human primary fibroblasts obtained from tunica albuginea of patients with Peyronie's disease were transformed to myofibroblasts using transforming growth factor beta 1 (TGF-β1). The mRNA of the cells was collected at 0, 24, 36, 48, and 72 hours after stimulation with TGF-β1 and then analyzed using a Nanostring nCounter Fibrosis panel. The gene expression results were analyzed using Reactome pathway analysis database and ANNi, a deep learning-based inference algorithm based on a swarm approach.
    Outcomes: The study outcome was the time course of changes in gene expression during transformation of PD-derived fibroblasts to myofibroblasts.
    Results: The temporal analysis of the gene expression revealed that the majority of the changes at the gene expression level happened within the first 24 hours and remained so throughout the 72-hour period. At 36 hours, significant changes were observed in genes involved in MAPK-Hedgehog signaling pathways.
    Clinical translation: This study highlights the importance of early intervention in clinical management of PD and the future potential of new drugs targeting the point of no return.
    Strengths and limitations: The use of human primary cells and confirmation of results with further RNA analysis are the strengths of this study. The study was limited to 760 genes rather than the whole transcriptome.
    Conclusion: This study is to our knowledge the first analysis of temporal gene expression associated with the regulation of the transformation of resident fibroblasts to profibrotic myofibroblasts in PD. Further research is warranted to investigate the role of the MAPK-Hedgehog signaling pathways in reversibility of PD.
    MeSH term(s) Male ; Humans ; Penile Induration/genetics ; Myofibroblasts/metabolism ; Transforming Growth Factor beta1/metabolism ; Hedgehog Proteins/metabolism ; Penis ; Cells, Cultured ; Fibroblasts/metabolism
    Chemical Substances Transforming Growth Factor beta1 ; Hedgehog Proteins
    Language English
    Publishing date 2024-04-03
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 2251959-2
    ISSN 1743-6109 ; 1743-6095
    ISSN (online) 1743-6109
    ISSN 1743-6095
    DOI 10.1093/jsxmed/qdae006
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 6417: Show issues Location:
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    Zs.MO 505: Show issues

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  10. Article: The reproducibility of biomedical research: Sleepers awake!

    Bustin, Stephen A

    Biomolecular detection and quantification

    2015  Volume 2, Page(s) 35–42

    Abstract: There is increasing concern about the reliability of biomedical research, with recent articles suggesting that up to 85% of research funding is wasted. This article argues that an important reason for this is the inappropriate use of molecular techniques, ...

    Abstract There is increasing concern about the reliability of biomedical research, with recent articles suggesting that up to 85% of research funding is wasted. This article argues that an important reason for this is the inappropriate use of molecular techniques, particularly in the field of RNA biomarkers, coupled with a tendency to exaggerate the importance of research findings.
    Language English
    Publishing date 2015-01-21
    Publishing country Germany
    Document type Review ; Journal Article
    ZDB-ID 2821770-6
    ISSN 2214-7535
    ISSN 2214-7535
    DOI 10.1016/j.bdq.2015.01.002
    Database MEDical Literature Analysis and Retrieval System OnLINE

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