Optimization of Coated Blade Spray for Rapid Screening and Quantitation of 105 Veterinary Drugs in Biological Tissue Samples

Abstract Background Grass pea (Lathyrus sativus) is an underutilised crop with high tolerance to drought and flooding stress and potential for maintaining food and nutritional security in the face of climate change. The presence of the neurotoxin β-L-oxalyl-2,3-diaminopropionic acid (β-L-ODAP) in tissues of the plant has limited its adoption as a staple crop. To assist in the detection of material with very low neurotoxin toxin levels, we have developed two novel methods to assay ODAP. The first, a version of a widely used spectrophotometric assay, modified for increased throughput, permits rapid screening of large populations of germplasm for low toxin lines and the second is a novel, mass spectrometric procedure to detect very small quantities of ODAP for research purposes and characterisation of new varieties. Results A plate assay, based on an established spectrophotometric method enabling high-throughput ODAP measurements, is described. In addition, we describe a novel liquid chromatography mass spectrometry (LCMS)-based method for β-L-ODAP-quantification. This method utilises an internal standard (di-13C-labelled β-L-ODAP) allowing accurate quantification of β-L-ODAP in grass pea tissue samples. The synthesis of this standard is also described. The two methods are compared; the spectrophotometric assay lacked sensitivity and detected ODAP-like absorbance in chickpea and pea whereas the LCMS method did not detect any β-L-ODAP in these species. The LCMS method was also used to quantify β-L-ODAP accurately in different tissues of grass pea. Conclusions The plate-based spectrophotometric assay allows quantification of total ODAP in large numbers of samples, but its low sensitivity and inability to differentiate α- and β-L-ODAP limit its usefulness for accurate quantification in low-ODAP samples. Coupled to the use of a stable isotope internal standard with LCMS that allows accurate quantification of β-L-ODAP in grass pea samples with high sensitivity, these methods permit the identification and characterisation of grass pea lines with a very low ODAP content. The LCMS method is offered as a new ‘gold standard’ for β-L-ODAP quantification, especially for the validation of existing and novel low- and/or zero-β-L-ODAP genotypes.

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Laboratory Specimens and Genetic Privacy: Evolution of Legal Theory

The Journal of Law Medicine & Ethics ◽

10.1111/jlme.12042 ◽

2013 ◽

Vol 41 (S1) ◽

pp. 65-68 ◽

Cited By ~ 1

Author(s):

Michelle Huckaby Lewis

Keyword(s):

Biomedical Research ◽

Biological Samples ◽

Biological Tissue ◽

Genetic Information ◽

Legal Theory ◽

Genetic Privacy ◽

Tissue Samples

Human biological tissue samples are an invaluable resource for biomedical research designed to find causes of diseases and their treatments. Controversy has arisen, however, when research has been conducted with laboratory specimens either without the consent of the source of the specimen or when the research conducted with the specimen has expanded beyond the scope of the original consent agreement. Moreover, disputes have arisen regarding which party, the researcher or the source of the specimen, has control over who may use the specimens and for what purposes. The purposes of this article are: (1) to summarize the most important litigation regarding the use of laboratory specimens, and (2) to demonstrate how legal theory regarding control of laboratory specimens has evolved from arguments based upon property interests in biological samples to claims that the origins of laboratory specimens have privacy interests in their genetic information that should be protected.

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Serial Block-Face Scanning Electron Microscopy (SBF-SEM) of Biological Tissue Samples

Journal of Visualized Experiments ◽

10.3791/62045 ◽

2021 ◽

Author(s):

Justin A. Courson ◽

Paul T. Landry ◽

Thao Do ◽

Eric Spehlmann ◽

Pascal J. Lafontant ◽

...

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Biological Tissue ◽

Tissue Samples ◽

Face Scanning ◽

Block Face ◽

Scanning Electron

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An image formation model for Secondary Ion Mass Spectrometry imaging of biological tissue samples

Applied Surface Science ◽

10.1016/j.apsusc.2010.08.046 ◽

2010 ◽

Vol 257 (4) ◽

pp. 1267-1275 ◽

Cited By ~ 2

Author(s):

Gaia Volandri ◽

Luca Menichetti ◽

Marco Matteucci ◽

Claudia Kusmic ◽

Marco Consumi ◽

...

Keyword(s):

Mass Spectrometry ◽

Biological Tissue ◽

Mass Spectrometry Imaging ◽

Secondary Ion Mass Spectrometry ◽

Image Formation ◽

Formation Model ◽

Tissue Samples ◽

Ion Mass Spectrometry ◽

Secondary Ion

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Microfluidic platforms for rapid screening of cancer affinity reagents by using tissue samples

Biomicrofluidics ◽

10.1063/1.5050451 ◽

2018 ◽

Vol 12 (5) ◽

pp. 054108 ◽

Cited By ~ 4

Author(s):

Lien-Yu Hung ◽

Chien-Yu Fu ◽

Chih-Hung Wang ◽

Yuan-Jhe Chuang ◽

Yi-Cheng Tsai ◽

...

Keyword(s):

Rapid Screening ◽

Microfluidic Platforms ◽

Tissue Samples ◽

Affinity Reagents

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Spatiotemporal Characterizations of Spontaneously Beating Cardiomyocytes with Adaptive Reference Digital Image Correlation

Scientific Reports ◽

10.1038/s41598-019-54768-w ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Akankshya Shradhanjali ◽

Brandon D. Riehl ◽

Bin Duan ◽

Ruiguo Yang ◽

Jung Yul Lim

Keyword(s):

Regenerative Medicine ◽

Digital Image Correlation ◽

Digital Image ◽

Biological Tissue ◽

Image Correlation ◽

Myocardial Tissue ◽

Tissue Development ◽

Tissue Samples ◽

Non Invasive ◽

Spatiotemporal Parameters

AbstractWe developed an Adaptive Reference-Digital Image Correlation (AR-DIC) method that enables unbiased and accurate mechanics measurements of moving biological tissue samples. We applied the AR-DIC analysis to a spontaneously beating cardiomyocyte (CM) tissue, and could provide correct quantifications of tissue displacement and strain for the beating CMs utilizing physiologically-relevant, sarcomere displacement length-based contraction criteria. The data were further synthesized into novel spatiotemporal parameters of CM contraction to account for the CM beating homogeneity, synchronicity, and propagation as holistic measures of functional myocardial tissue development. Our AR-DIC analyses may thus provide advanced non-invasive characterization tools for assessing the development of spontaneously contracting CMs, suggesting an applicability in myocardial regenerative medicine.

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Development of a Genus-Specific Brucella Real-Time PCR Assay Targeting the 16S-23S rDNA Internal Transcribed Spacer from Different Specimen Types

Veterinary Sciences ◽

10.3390/vetsci7040175 ◽

2020 ◽

Vol 7 (4) ◽

pp. 175

Author(s):

Rejoice Nyarku ◽

Ayesha Hassim ◽

Annelize Jonker ◽

Melvyn Quan

Keyword(s):

Sensitivity And Specificity ◽

Internal Transcribed Spacer ◽

Bacterial Culture ◽

Limit Of Detection ◽

Quantitative Polymerase Chain Reaction ◽

Surface Protein ◽

Qpcr Assay ◽

Rapid Screening ◽

Analytical Sensitivity ◽

Tissue Samples

The aim of this study was to develop a 16S-23S ribosomal deoxyribonucleic acid internal transcribed spacer (ITS) quantitative polymerase chain reaction (qPCR) assay for the early diagnosis and rapid screening of brucellosis. Blood, milk, and tissue samples were spiked with B. abortus biovar 1 (B01988-18 strain) to determine the analytical sensitivity and specificity of the assay. The 95% limit of detection of the ITS qPCR assay was highest in tissue, followed by blood, then milk, i.e., 0.48, 4.43, and 15.18 bacteria/PCR reaction, respectively. The diagnostic performance of the assay was compared to the Brucella cell surface protein (BCSP) 31 qPCR assay and bacterial culture. Out of 56 aborted foetal tissue samples from bovine, ovine, and caprine, 33% (19/56) were positive for Brucella spp. The sensitivity and specificity of the ITS qPCR assay was 87% and 95% respectively, compared to 92% and 89% for the BCSP31 qPCR assay and 47% and 55% for bacterial culture, respectively. The assay was efficient, sensitive, and specific, making it a valuable tool in the early detection of the Brucella pathogen.

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