Comparison of Tissue Preparation Methods for Assay of Nicotinamide Coenzymes

Zhao, Zhifan; Hu, Xiuchan; Ross, Cleon W.

doi:10.1104/pp.84.4.987

Cited by 41 publications

(22 citation statements)

References 5 publications

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“…The bioassay utilized relies on NADH reduction of formazan (MTT) reagent. The intensity of the reduced MTT compound, measured at 565 nm, is proportional to the NAD ϩ -to-NADH ratio (37,56). Consistent with our other findings, chronic ethanol consumption significantly decreased the mitochondrial NAD ϩ -to-NADH ratio (Control: 1.0 Ϯ 0.11, Ethanol: 0.39 Ϯ 0.06; P Ͻ 0.05 from 6 independent animals studied).…”

Section: Effect Of Lps On Alveolar Fluid Clearance In the Chronic Alcsupporting

confidence: 90%

Chronic ethanol exposure alters the lung proteome and leads to mitochondrial dysfunction in alveolar type 2 cells

Alli

Brewer

Montgomery

et al. 2014

American Journal of Physiology-Lung Cellular and Molecular Phys

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The lungs can undergo irreversible damage from chronic alcohol consumption. Herein, we developed an animal model predisposed for edematous lung injury following chronic ingestion of alcohol to better understand the etiology of alcohol-related disorders. Using animal modeling, alongside high-throughput proteomic and microarray assays, we identified changes in lung protein and transcript in mice and rats, respectively, following chronic alcohol ingestion or a caloric control diet. Liquid chromatography-mass spectrometry identified several mitochondrial-related proteins in which the expression was upregulated following long-term alcohol ingestion in mice. Consistent with these observations, rat gene chip microarray analysis of alveolar cells obtained from animals maintained on a Lieber-DeCarli liquid alcohol diet confirmed significant changes in mitochondrial-related transcripts in the alcohol lung. Transmission electron microscopy revealed significant changes in the mitochondrial architecture in alcohol mice, particularly following lipopolysaccharide exposure. Chronic alcohol ingestion was also shown to worsen mitochondrial respiration, mitochondrial membrane polarization, and NAD(+)-to-NADH ratios in alveolar type 2 cells. In summary, our studies show causal connection between chronic alcohol ingestion and mitochondrial dysfunction, albeit the specific role of each of the mitochondrial-related proteins and transcripts identified in our study requires additional study.

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Section: Effect Of Lps On Alveolar Fluid Clearance In the Chronic Alcsupporting

confidence: 90%

Chronic ethanol exposure alters the lung proteome and leads to mitochondrial dysfunction in alveolar type 2 cells

Alli

Brewer

Montgomery

et al. 2014

American Journal of Physiology-Lung Cellular and Molecular Phys

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show abstract

“…Radicles (c. 5 mg dry mass) were homogenized in 5ml of 0.2 M HCl following homogenization to a fi ne powder under liquid nitrogen in the mortar with a pestle, heated in a boiling water bath for 5 min, cooled in an ice bath and centrifuged at 14 000 g for 10 min (Zhao et al 1987). An aliquot of 0.5 ml of the supernatant was then transferred to 1.0 M Bicine-NaOH buff er (pH 8.0) and neutralised with 0.2 M NaOH in the dark (Matsumura and Miyachi, 1980).…”

Section: Oxidised Nicotinamide Adenine Dinucleotide (Nad) Assaymentioning

confidence: 99%

Increasing the rate of drying reduces metabolic imbalance, lipid peroxidation and critical water content in radicles of garden pea (Pisum sativum L.)

Pammenter

Berjak

2013

Biol. Res.

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Orthodox seeds become desiccation-sensitive as they undergo germination. As a result, germinating seeds serve as a model to study desiccation sensitivity in plant tissues. The eff ects of the rate of drying on the viability, respiratory metabolism and free radical processes were thus studied during dehydration and wet storage of radicles of Pisum sativum. For both drying regimes desiccation could be described by exponential and inverse modifi ed functions. Viability, as assessed by germination capacity and tetrazolium staining, remained at 100% during rapid (< 24 h) desiccation. However, it declined sharply at c. 0.26 g g -1 dm following slow (c. 5 days) drying. Increasing the rate of dehydration thus lowered the critical water content for survival. Rapid desiccation was also associated with higher activities and levels of malate dehydrogenase and the oxidized form of nicotinamide adenine dinucleotide. It was also accompanied by lower hydroperoxide levels and membrane damage. In addition, the activitiy of glutathione reductase was greater during rapid drying. Ageing may have contributed to increased damage during slow dehydration, since viability declined even in wet storage after two weeks. The results presented are consistent with rapid desiccation reducing the accumulation of damage resulting from desiccation-induced aqueous-based deleterious reactions. In addition, they show that radicles are a useful model to study desiccation sensitivity in plant tissues.

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“…Reduced (NADPH) and oxidized (NADP ' ) forms were extracted from the post mitochondrial fraction according to the method of Zhao et al (1987). Enzyme cycling assay was employed as following the procedure described by Matsumura and Miyachi (1980) for the coenzyme quantification.…”

Section: Nadph/nadp Determinationmentioning

confidence: 99%

Cadmium affects the NADP-thioredoxin reductase/thioredoxin system in germinating pea seeds

Smiri

Jelali

Ghoul

2013

Journal of Plant Interactions

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Seeds have devised sophisticated mechanisms to cope with adverse situations. Among these mechanisms the collectively referred to as redox homeostasis. We have investigated the thioredoxin (Trx)/NADPH/NADPHdependent thioredoxin reductase (NTR) systems during the pea (Pisum sativum L.) seed germination under heavy metal stress. Cotyledon and embryonic axis were analyzed between 0 and 5 days of germination in the presence or absence of cadmium (Cd). Under Cd stress conditions, NTR activity and Trx h3 and Trx h4 expression were stimulated, while the overall activity of Trx decreased. When incubated with Cd ions in vitro, the disulfide reductase activity of Trx h3 and f isoforms was drastically inhibited, probably through the formation of protein dimers as evidenced by the electrophoresis analysis. The NADPH status was also affected, since Cd treatment provoked an increase in oxidized state of coenzyme as compared to control redox ratio. The contribution of Trx system to the redox regulation during the germination is discussed in relation with the Cd-caused oxidative stress.

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Comparison of Tissue Preparation Methods for Assay of Nicotinamide Coenzymes

Cited by 41 publications

References 5 publications

Chronic ethanol exposure alters the lung proteome and leads to mitochondrial dysfunction in alveolar type 2 cells

Chronic ethanol exposure alters the lung proteome and leads to mitochondrial dysfunction in alveolar type 2 cells

Increasing the rate of drying reduces metabolic imbalance, lipid peroxidation and critical water content in radicles of garden pea (Pisum sativum L.)

Cadmium affects the NADP-thioredoxin reductase/thioredoxin system in germinating pea seeds

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