Cheap indicator
Russian scientists have developed a test for biological age
Scientists from Moscow State University have proposed a new method for determining the level of one of the compounds-indicators of a person's biological age using an inexpensive set of chemical reagents. The results of the study are published in the journal Analytical Biochemistry (Artiukhov et al., Advantages of formate dehydrogenase reaction for efficient NAD+ quantification in biological samples).
Nicotinamide adenine dinucleotide (NAD) is a compound used by living beings to generate energy. In metabolic processes, it takes electrons from some molecules, oxidizing them, and gives them to others, restoring them. Recently, the functions of this compound associated with chemical modifications of proteins have been discovered.
Scientists from Moscow State University have proposed using the NAD+ level as one of the markers of the biological age of the body and pathologies of its metabolic processes, such as obesity, some heart and liver diseases, neurodegenerative diseases, for example, Alzheimer's and Parkinson's diseases, chronic inflammation and cancer. In addition, the content of NAD+ can serve as an indicator of the effectiveness of the pharmacological action of drugs aimed at combating these diseases. Research supported by a grant The Russian Science Foundation (RNF).
Scientists have found out that the NAD form, capable of taking electrons, participates in the work of the cell's signaling systems, allowing it to respond to external stimuli. Changes in the amount of NAD+ in cells during the day are involved in the mechanisms of maintaining the biological clock – the so-called circadian rhythms. The work of the biological clock deteriorates during aging, and its violation leads to various diseases.
Having easily accessible, fast and inexpensive methods for determining the amount of NAD+ in cells, this indicator can be used in clinical and research practice. However, the methods currently available require sophisticated equipment, highly qualified personnel and expensive consumables, which limits their applicability for clinical analyses and implementation into practice.
As an alternative approach, the staff of the A. N. Belozersky Research Institute of Physico-Chemical Biology of Moscow State University and colleagues proposed using formate dehydrogenase (FDG), an enzyme that microorganisms and plants have. It oxidizes the smallest of the organic acids, formic acid, transferring its electrons to the NAD+ molecule in the form of a hydride ion - a negatively charged hydrogen ion containing an "extra" electron.
In this case, NAD+ passes from the oxidized form to the reduced one, which is able to fluoresce. Due to this, the amount of electrons received by NAD+ can be determined using fluorescence analysis. Such properties make it possible to use FDG for the determination of NAD+ in complex samples of biological fluids containing many different compounds.
The researchers propose to create a commercial set of reagents for such enzymatic measurements of NAD+ in the blood of patients. Analyses can be carried out on microplate readers – relatively inexpensive devices widely used for diagnostics, biotechnology, veterinary medicine and scientific research. A tablet with many holes is placed in the device, which allows you to simultaneously analyze up to several hundred samples.
"Analogues of commercial kits available from foreign manufacturers for measuring the content of NAD + use enzymes that have a number of significant drawbacks that are absent from the one used in our work. In addition to the high efficiency and specificity of NAD+ binding, FDG has high physical and chemical resistance. Therefore, one batch of the drug can be used to measure a large number of different biological samples for a long time," the words of the head of the study, Doctor of Chemical Sciences Victoria Bunik, professor of the Faculty of Bioengineering and Bioinformatics of Lomonosov Moscow State University and the First Sechenov Moscow State Medical University are quoted in the RNF press release.
The authors believe that the proposed method can significantly simplify and reduce the cost of current methods of measuring the level of NAD+, which will help their implementation in clinical and research practice.
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