Biochemical paths in humans and cells: Frontiers of AMS bioanalysis

Vogel, John S.; Palmblad, Magnus; Ognibene, Ted; Kabir, Md. Mohiuddin; Buchholz, Bruce A.; Bench, Graham

doi:10.1016/j.nimb.2007.01.215

Cited by 12 publications

(7 citation statements)

References 41 publications

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“…NADH has been shown to function as an inhibitor of Sir2 [10], so lower levels of NADH found in yeast cells grown under CR could result in less inhibition of Sir2 and therefore an increased lifespan in yeast [10]. By culturing yeast in 14 C-labeled growth media, we are investigating the relative contributions of the de novo and salvage pathways to total NAD and NADH levels in yeast strains as a function of nutrient status [40]. The protocol developed here avoids derivitization or the measurement of degradation products from NAD or NADH so that the potential for loss of the isotope label is minimized.…”

Section: Discussionmentioning

confidence: 99%

“…The protocol developed here avoids derivitization or the measurement of degradation products from NAD or NADH so that the potential for loss of the isotope label is minimized. Initial radiolabeling of yeast cells and subsequent accelerator MS (AMS) measurement for 14 C quantitation indicate that: (i) yeast cells are amenable to such labeling and quantification [40]; and (ii) the single sample extraction and HPLC speciation protocol developed here are amenable to AMS quantitation of 14 C in isolated fractions of the HPLC trace [40] enabling us to quantify total NAD and NADH (by UV–Vis absorption) and the contribution of NAD and NADH from a specific pathway (by AMS) from the same sample.…”

Section: Discussionmentioning

confidence: 99%

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Single sample extraction protocol for the quantification of NAD and NADH redox states in Saccharomyces cerevisiae

Sporty¹,

Kabir²,

Turteltaub³

et al. 2008

J of Separation Science

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A robust redox extraction protocol for quantitative and reproducible metabolite isolation and recovery has been developed for simultaneous measurement of nicotin-amide adenine dinucleotide (NAD) and its reduced form, NADH, from Saccharomyces cerevisiae. Following culture in liquid media, yeast cells were harvested by centrifugation and then lysed under nonoxidizing conditions by bead blasting in ice-cold, nitrogen-saturated 50 mM ammonium acetate. To enable protein denaturation, ice cold nitrogen-saturated CH 3 CN/50 mM ammonium acetate (3:1 v/v) was added to the cell lysates. Chloroform extractions were performed on supernatants to remove organic solvent. Samples were lyophilized and resuspended in 50 mM ammonium acetate. NAD and NADH were separated by HPLC and quantified using UV-Vis absorbance detection. NAD and NADH levels were evaluated in yeast grown under normal (2% glucose) and calorie restricted (0.5% glucose) conditions. Results demonstrate that it is possible to perform a single preparation to reliably and robustly quantitate both NAD and NADH contents in the same sample. Robustness of the protocol suggests it will be (i) applicable to quantification of these metabolites in other cell cultures; and (ii) amenable to isotope labeling strategies to determine the relative contribution of specific metabolic pathways to total NAD and NADH levels in cell cultures.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Single sample extraction protocol for the quantification of NAD and NADH redox states in Saccharomyces cerevisiae

Sporty¹,

Kabir²,

Turteltaub³

et al. 2008

J of Separation Science

View full text Add to dashboard Cite

show abstract

“…AMS’s complexity, large size, time consuming sample processing, and relatively high cost have been an obstacle to the scientific community’s adoption of the method and its applications [23]. This has led to several scientific groups exploring new ways to measure 14 C [24,25,26,27,28].…”

Section: Technologymentioning

confidence: 99%

Radiocarbon Tracers in Toxicology and Medicine: Recent Advances in Technology and Science

Malfatti

Buchholz

Enright

et al. 2019

Toxics

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This review summarizes recent developments in radiocarbon tracer technology and applications. Technologies covered include accelerator mass spectrometry (AMS), including conversion of samples to graphite, and rapid combustion to carbon dioxide to enable direct liquid sample analysis, coupling to HPLC for real-time AMS analysis, and combined molecular mass spectrometry and AMS for analyte identification and quantitation. Laser-based alternatives, such as cavity ring down spectrometry, are emerging to enable lower cost, higher throughput measurements of biological samples. Applications covered include radiocarbon dating, use of environmental atomic bomb pulse radiocarbon content for cell and protein age determination and turnover studies, and carbon source identification. Low dose toxicology applications reviewed include studies of naphthalene-DNA adduct formation, benzo[a]pyrene pharmacokinetics in humans, and triclocarban exposure and risk assessment. Cancer-related studies covered include the use of radiocarbon-labeled cells for better defining mechanisms of metastasis and the use of drug-DNA adducts as predictive biomarkers of response to chemotherapy.

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“…The use of man-made radioisotopes in the field of medicine for research, diagnostics and therapy is well known. AMS plays an important role in biomedical research through its outstanding detection sensitivity of long-lived radioisotopes [135]. The very small amount of radioisotopes required for testing new drugs allowed one to study the metabolism and pharmacokinetics of these drugs by microdosing of humans, considerably reducing the time for a new drug development [136].…”

Section: Biomedical Research With Radioisotopesmentioning

confidence: 99%

Accelerator mass spectrometry: state of the art and perspectives

Kutschera

2016

Advances in Physics: X

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Accelerator mass spectrometry (AMS) is sometimes called 'the art of counting atoms one by one'. In addition to counting individual atoms, AMS is also capable to determine both mass number (A) and atomic number (Z). Since an atom (also called a nuclide) is unambiguously characterized by the proton number (Z) and the neutron number (N = A − Z), a rare nuclide can be well separated from possible background events, and extremely low abundances of specific nuclides can be measured. In general, the use of an accelerator system as a mass spectrometer improves the isotope abundance sensitivity by many orders of magnitude as compared to standard mass spectrometry (without an accelerator). In particular, AMS provides the means to measure minute traces of long-lived radioisotopes of cosmogenic and/or anthropogenic origin in essentially every domain of our environment at large. This allows one to use AMS for performing research in many different areas, ranging from archaeology to astrophysics. In this review, an update of the current status of AMS and an outlook to future developments in both technical and applied aspects will be presented. The transition from Mass Spectrometry (MS) to Accelera tor Mass Spectrometry (AMS)

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Biochemical paths in humans and cells: Frontiers of AMS bioanalysis

Cited by 12 publications

References 41 publications

Single sample extraction protocol for the quantification of NAD and NADH redox states in Saccharomyces cerevisiae

Single sample extraction protocol for the quantification of NAD and NADH redox states in Saccharomyces cerevisiae

Radiocarbon Tracers in Toxicology and Medicine: Recent Advances in Technology and Science

Accelerator mass spectrometry: state of the art and perspectives

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