Biochemistry (Moscow) (v.81, #12)

Mitochondrial genome and longevity by R. A. Zinovkin; M. V. Skulachev; V. P. Skulachev (1401-1405).
The mitochondrial genome provides not only respiratory chain function, but it also ensures the impact of mitochondria on nearly all crucial metabolic processes. It is well known that mitochondria regulate aging and lifespan. However, until now there were no direct experimental data concerning the influence of various mitochondrial DNA variants on lifespan of animals with identical nuclear genome. In a recent paper of J. A. Enriquez and coworkers (Latorre-Pellicer, A., et al. (2016) Nature, 535, 561-565), it was shown that mice carrying nuclear DNA from one strain and mitochondrial DNA from another had longer median lifespan and retarded development of various aging traits. This review critically analyzes that paper and considers some aspects of the crosstalk between the nuclear and mitochondrial genomes. We also discuss new perspectives of gerontology in the light of the discovery made by Enriquez’s group.
Keywords: mitochondria; mitochondrial DNA; nuclear genome; lifespan; aging

Programmed aging refers to the idea that senescence in humans and other organisms is purposely caused by evolved biological mechanisms to obtain an evolutionary advantage. Until recently, programmed aging was considered theoretically impossible because of the mechanics of the evolution process, and medical research was based on the idea that aging was not programmed. Theorists struggled for more than a century in efforts to develop non-programmed theories that fit observations, without obtaining a consensus supporting any non-programmed theory. Empirical evidence of programmed lifespan limitations continued to accumulate. More recently, developments, especially in our understanding of biological inheritance, have exposed major issues and complexities regarding the process of evolution, some of which explicitly enable programmed aging of mammals. Consequently, science-based opposition to programmed aging has dramatically declined. This progression has major implications for medical research, because the theories suggest that very different biological mechanisms are ultimately responsible for highly age-related diseases that now represent most research efforts and health costs. Most particularly, programmed theories suggest that aging per se is a treatable condition and suggest a second path toward treating and preventing age-related diseases that can be exploited in addition to the traditional disease-specific approaches. The theories also make predictions regarding the nature of biological aging mechanisms and therefore suggest research directions. This article discusses developments of evolutionary mechanics, the consequent programmed aging theories, and logical inferences concerning biological aging mechanisms. It concludes that major medical research organizations cannot afford to ignore programmed aging concepts in assigning research resources and directions.
Keywords: medical research policy; aging theories; senescence; programmed aging; evolution

Possible interventions to modify aging by Giacinto Libertini; Nicola Ferrara (1413-1428).
The programmed aging paradigm interprets aging as a function favored by natural selection at a supra-individual level. This function is implemented, according to the telomere theory, through mechanisms that operate through the subtelomere–telomere–telomerase system. After reviewing some necessary technical and ethical reservations and providing a concise description of aging mechanisms, this work considers interventions that could lead to the control of some highly disabling characteristics of aging, such as Alzheimer’s and Parkinson’s syndromes and age-related macular degeneration, and afterwards to a full control of aging up to a condition equivalent to that of the species defined as “with negligible senescence”. The various steps needed for the development of such interventions are described along general lines.
Keywords: aging; programmed aging paradigm; Alzheimer; Parkinson; telomere; subtelomere

Study of age-dependent structural and functional changes of mitochondria in skeletal muscles and heart of naked mole rats (Heterocephalus glaber) by S. Holtze; C. M. Eldarov; V. B. Vays; I. M. Vangeli; M. Yu. Vysokikh; L. E. Bakeeva; V. P. Skulachev; T. B. Hildebrandt (1429-1437).
Morphometric analysis of mitochondria in skeletal muscles and heart of 6- and 60-month-old naked mole rats (Heterocephalus glaber) revealed a significant age-dependent increase in the total area of mitochondrial cross-sections in studied muscle fibers. For 6- and 60-month-old animals, these values were 4.8 ± 0.4 and 12.7 ± 1.8%, respectively. This effect is mainly based on an increase in the number of mitochondria. In 6-month-old naked mole rats, there were 0.23 ± 0.02 mitochondrial cross-sections per μm2 of muscle fiber, while in 60-month-old animals this value was 0.47 ± 0.03. The average area of a single mitochondrial cross-section also increased with age in skeletal muscles–from 0.21 ± 0.01 to 0.29 ± 0.03 μm2. Thus, naked mole rats show a drastic enlargement of the mitochondrial apparatus in skeletal muscles with age due to an increase in the number of mitochondria and their size. They possess a neotenic type of chondriome accompanied by specific features of mitochondrial functioning in the state of oxidative phosphorylation and a significant decrease in the level of matrix adenine nucleotides.
Keywords: naked mole rat; morphometry; respiration; aging; ultrastructure; mitochondria; neoteny

Uncouplers of oxidation and phosphorylation as antiaging compounds by D. A. Knorre; F. F. Severin (1438-1444).
Food restriction causes a set of physiological changes that reduce the rate of aging. At the level of an organism, these changes are initiated by a hormonal response, which in turn activates certain intracellular signaling cascades. As a result, cells increase their antioxidant capacities and decrease the risk of cancerous transformation. A number of small molecule compounds activating these signaling cascades have been described. One could expect that direct pharmacological activation of the signaling can produce a stronger antiaging effect than that achieved by the indirect hormonal stimulation. Data from the literature point to the opposite. Possibly, a problem with pharmacological activators is that they cause generation of mitochondrial reactive oxygen species. Indeed, hyperpolarized mitochondria are known to induce oxidative stress. Such hyperpolarization could happen because of artificial activation of cellular response to caloric restriction in the absence of energy deficit. At the same time, energy deficit seems likely to be a natural consequence of the shortage of nutrients. Thus, there is a possibility that combining the pharmacological activators with compounds that decrease mitochondrial transmembrane potential, uncouplers, could be a powerful antiaging strategy.
Keywords: aging; food restriction; caloric restriction; uncouplers; geroprotectors; mTOR; AMPK

Experimental adaptation of Drosophila melanogaster to nutrient-deficient starch-based (S) medium resulted in lifespan shortening, increased early-life fecundity, accelerated reproductive aging, and sexually dimorphic survival curves. The direction of all these evolutionary changes coincides with the direction of phenotypic plasticity observed in non-adapted flies cultured on S medium. High adult mortality rate caused by unfavorable growth medium apparently was the main factor of selection during the evolutionary experiment. The results are partially compatible with Williams’ hypothesis, which states that increased mortality rate should result in relaxed selection against mutations that decrease fitness late in life, and thus promote the evolution of shorter lifespan and earlier reproduction. However, our results do not confirm Williams’ prediction that the sex with higher mortality rate should undergo more rapid aging: lifespan shortening by S medium is more pronounced in naive males than females, but it was female lifespan that decreased more in the course of adaptation. These data, as well as the results of testing of F1 hybrids between adapted and control lineages, are compatible with the idea that the genetic basis of longevity is different in the two sexes, and that evolutionary response to increased mortality rate depends on the degree to which the mortality is selective. Selective mortality can result in the development of longer (rather than shorter) lifespan in the course of evolution. The results also imply that antagonistic pleiotropy of alleles, which increase early-life fecundity at the cost of accelerated aging, played an important role in the evolutionary changes of females in the experimental lineage, while accumulation of deleterious mutations with late-life effects due to drift was more important in the evolution of male traits.
Keywords: experimental evolution; adaptation; lifespan; fecundity; aging; life cycle evolution; evolutionary trade-off

Is it possible to prove the existence of an aging program by quantitative analysis of mortality dynamics? by G. A. Shilovsky; T. S. Putyatina; S. N. Lysenkov; V. V. Ashapkin; O. S. Luchkina; A. V. Markov; V. P. Skulachev (1461-1476).
Accumulation of various types of lesions in the course of aging increases an organism’s vulnerability and results in a monotonous elevation of mortality rate, irrespective of the position of a species on the evolutionary tree. Stroustrup et al. (Nature, 530, 103–107) [1] showed in 2016 that in the nematode Caenorhabditis elegans, longevity-altering factors (e.g. oxidative stress, temperature, or diet) do not change the shape of the survival curve, but either stretch or shrink it along the time axis, which the authors attributed to the existence of an “aging program”. Modification of the accelerated failure time model by Stroustrup et al. uses temporal scaling as a basic approach for distinguishing between quantitative and qualitative changes in aging dynamics. Thus we analyzed data on the effects of various longevity-increasing genetic manipulations in flies, worms, and mice and used several models to choose a theory that would best fit the experimental results. The possibility to identify the moment of switch from a mortality-governing pathway to some other pathways might be useful for testing geroprotective drugs. In this work, we discuss this and other aspects of temporal scaling.
Keywords: lifespan; aging; survival curves; temporal scaling; phenoptosis

Effect of alpha-fetoprotein on lifespan of old mice by V. N. Krut’ko; V. I. Dontsov; A. V. Khalyavkin (1477-1479).
Alpha-fetoprotein (AFP) is one of the best-known embryo-specific proteins. It is used to diagnose fetal abnormalities and tumors of the gastrointestinal tract and liver. AFP has pronounced immunotropic and detoxifying effect and a direct apoptotic effect on tumor cells. The treatment of mice at the oldest age in our experiments with AFP dramatically increased the survival and markedly increased the relative weight of immunotropic organs, apparently due to the general effect of AFP in improving functions of tissues and detoxifying actions. It also improved appearance and the relative weight of internal organs with a reduced age of autoaggression.
Keywords: age; alpha-fetoprotein; life extension

Molecular and cellular interactions between mother and fetus. Pregnancy as a rejuvenating factor by V. A. Popkov; D. N. Silachev; S. S. Jankauskas; L. D. Zorova; I. B. Pevzner; V. A. Babenko; E. Y. Plotnikov; D. B. Zorov (1480-1487).
Aging is associated with a decline of various body functions, including ability to regenerate. Over recent decades, it has been demonstrated that some of these changes could be reversed in response to factors originating from a young organism, for example, fetal stem cells or “young blood” in models of heterochronic parabiosis. Pregnancy might be considered as parabiotic model of the interaction between two organisms of different age. In this work, we analyzed and summarized data on the effects of pregnancy on the maternal organism that confirm the hypothesis that pregnancy rejuvenates the mother’s organism or slows its aging.
Keywords: pregnancy; rejuvenation; aging

Mitochondria-targeted antioxidant SkQ1 did not increase the content of cytochromes P450 in livers of rats that were given SkQ1 in drinking water for 5 days in a dose (2.5 μmol per kg body weight) that exceeded 10 times the SkQ1 therapeutic dose. SkQ1 did not affect the levels of cytochrome P450 forms CYP1A2, CYP2B6, and CYP3A4 in monolayer cultures of freshly isolated human hepatocytes, while specific inducers of these forms (omeprazole, phenobarbital, and rifampicin, respectively) significantly increased expression of the cytochromes P450 under the same conditions. We conclude that therapeutic doses of SkQ1 do not induce cytochromes P450 in liver, and the absence of the inducing effect cannot be explained by poor availability of hepatocytes to SkQ1 in vivo.
Keywords: cytochrome P450 ; SkQ1; mitochondria-targeted antioxidant; hepatocyte culture

The vast majority of the Earth’s population lives between the 20th and 40th parallel north and south. It seems that right here humans have found the best living conditions relating not only to temperature and food recourses, but also to UV radiation necessary for the production of vitamin D by human skin. An exception to this general rule is Europe. Nearly half a billion people live between the 40th and 60th parallel north of the equator despite the fact that the amounts of UV radiation there are much lower. Moreover, since the time of the Vikings, there has always been a part of the European population that lived even further north than the 60th parallel (the northern parts of Europe, including Greenland). In this work, we present the potential role that vitamin D deficiency might have played in the extinction of the Vikings of Greenland. We analyze factors that contribute to the discrepancy between the theoretical distribution of areas with vitamin D deficiency and today’s reality, like the impact of civilization, religious traditions, as well as vitamin D supplementation in food products and as a biologically active dietary additive. The global migration of people on a scale and speed never seen before is now even more important for this discrepancy.
Keywords: vitamin D (deficiency); UV radiation; Europe; Greenland; Vikings; Scottish paradox

2-Oxo acid dehydrogenase complexes are important metabolic checkpoints functioning at the intercept of sugar and amino acid degradation. This review presents a short summary of architectural, catalytic, and regulatory principles of the complexes structure and function, based on recent advances in studies of well-characterized family members. Special attention is given to use of synthetic phosphonate and phosphinate analogs of 2-oxo acids as selective and efficient inhibitors of the cognate complexes in biological systems of bacterial, plant, and animal origin. We summarize our own results concerning the application of synthetic analogs of 2-oxo acids in situ and in vivo to reveal functional interactions between 2-oxo acid dehydrogenase complexes and other components of metabolic networks specific to different cells and tissues. Based on our study of glutamate excitotoxicity in cultured neurons, we show how a modulation of metabolism by specific inhibition of its key reaction may be employed to correct pathologies. This approach is further developed in our study on the action of the phosphonate analog of 2-oxoglutarate in animals. The study revealed that upregulation of 2-oxoglutarate dehydrogenase complex is involved in animal stress response and may provide increased resistance to damaging effects, underlying so-called preconditioning. The presented analysis of published data suggests synthetic inhibitors of metabolic checkpoints as promising tools to solve modern challenges of systems biology, metabolic engineering, and medicine.
Keywords: directed metabolic regulation; 2-oxo acid dehydrogenase complex; phosphonate or phosphinate analog of 2-oxo acids; stress resistance; tricarboxylic acid cycle; amino acid metabolism

Structural properties of potexvirus coat proteins detected by optical methods by P. I. Semenyuk; O. V. Karpova; A. L. Ksenofontov; N. O. Kalinina; E. N. Dobrov; V. V. Makarov (1522-1530).
It has been shown by X-ray analysis that cores of coat proteins (CPs) from three potexviruses, flexible helical RNA-containing plant viruses, have similar α-helical structure. However, this similarity cannot explain structural lability of potexvirus virions, which is believed to determine their biological activity. Here, we used circular dichroism (CD) spectroscopy in the far UV region to compare optical properties of CPs from three potexviruses with the same morphology and similar structure. CPs from Alternanthera mosaic virus (AltMV), potato aucuba mosaic virus (PAMV), and potato virus X (PVX) have been studied in a free state and in virions. The CD spectrum of AltMV virions was similar to the previously obtained CD spectrum of papaya mosaic virus (PapMV) virions, but differed significantly from the CD spectrum of PAMV virions. The CD spectrum of PAMV virions resembled in its basic characteristics the CD spectrum of PVX virions characterized by molar ellipticity that is abnormally low for α-helical proteins. Homology modeling of the CP structures in AltMV, PAMV, and PVX virions was based on the known high-resolution structures of CPs from papaya mosaic virus and bamboo mosaic virus and confirmed that the structures of the CP cores in all three viruses were nearly identical. Comparison of amino acid sequences of different potexvirus CPs and prediction of unstructured regions in these proteins revealed a possible correlation between specific features in the virion CD spectra and the presence of disordered N-terminal segments in the CPs.
Keywords: plant helical viruses; potexviruses; virion; coat protein; structure; optical methods

The symbiotic unicellular chlorophyte Desmodesmus sp. IPPAS-2014 capable of growth at extremely high CO2 levels prohibitive for most other microalgae is an interesting model for studies of CO2 tolerance mechanisms and a promising organism for CO2 biocapture. We studied the initial (0-60 min) phase of acclimation of this microalga to an abrupt decrease in pH of the medium sparged with air/20% CO2 mixture. Acclimation of the culture to these conditions was accompanied by a sharp decrease in photochemical activity of the chloroplast followed by its recovery with a characteristic time of 10-50 min. We hypothesize that acidification of the cultivation medium by dissolving CO2 plays a key role in the observed decrease in the photochemical activity. The possible role of photosynthetic apparatus tolerance to abrupt acidification in overall high tolerance of symbiotic microalgae to extremely high CO2 levels is discussed.
Keywords: acidification; chlorophyll fluorescence; stress; tolerance

Dysfunction of kidney endothelium after ischemia/reperfusion and its prevention by mitochondria-targeted antioxidant by S. S. Jankauskas; N. V. Andrianova; I. B. Alieva; A. N. Prusov; D. D. Matsievsky; L. D. Zorova; I. B. Pevzner; E. S. Savchenko; Y. A. Pirogov; D. N. Silachev; E. Y. Plotnikov; D. B. Zorov (1538-1548).
One of the most important pathological consequences of renal ischemia/reperfusion (I/R) is kidney malfunctioning. I/R leads to oxidative stress, which affects not only nephron cells but also cells of the vascular wall, especially endothelium, resulting in its damage. Assessment of endothelial damage, its role in pathological changes in organ functioning, and approaches to normalization of endothelial and renal functions are vital problems that need to be resolved. The goal of this study was to examine functional and morphological impairments occurring in the endothelium of renal vessels after I/R and to explore the possibility of alleviation of the severity of these changes using mitochondria-targeted antioxidant 10-(6′-plastoquinonyl)decylrhodamine 19 (SkQR1). Here we demonstrate that 40-min ischemia with 10-min reperfusion results in a profound change in the structure of endothelial cells mitochondria, accompanied by vasoconstriction of renal blood vessels, reduced renal blood flow, and increased number of endothelial cells circulating in the blood. Permeability of the kidney vascular wall increased 48 h after I/R. Injection of SkQR1 improves recovery of renal blood flow and reduces vascular resistance of the kidney in the first minutes of reperfusion; it also reduces the severity of renal insufficiency and normalizes permeability of renal endothelium 48 h after I/R. In in vitro experiments, SkQR1 provided protection of endothelial cells from death provoked by oxygen–glucose deprivation. On the other hand, an inhibitor of NO-synthases, L-nitroarginine, abolished the positive effects of SkQR1 on hemodynamics and protection from renal failure. Thus, dysfunction and death of endothelial cells play an important role in the development of reperfusion injury of renal tissues. Our results indicate that the major pathogenic factors in the endothelial damage are oxidative stress and mitochondrial damage within endothelial cells, while mitochondria-targeted antioxidants could be an effective tool for the protection of tissue from negative effects of ischemia.
Keywords: ischemia; kidney; Doppler; blood vessels; oxidative stress; mitochondria

Alterations in tear biochemistry associated with postanesthetic chronic dry eye syndrome by E. Yu. Zernii; M. O. Golovastova; V. E. Baksheeva; E. I. Kabanova; I. E. Ishutina; O. S. Gancharova; A. E. Gusev; M. S. Savchenko; A. P. Loboda; L. F. Sotnikova; A. A. Zamyatnin Jr.; P. P. Philippov; I. I. Senin (1549-1557).
Perioperative dry eye syndrome (DES) is a common ocular complication of long-term general anesthesia. Chronic DES can lead to permanent damage to the cornea and disturbance of visual function, up to total loss of vision. Here, a relationship between the duration of general anesthesia and the risk of chronic DES in patients was demonstrated. Using an experimental model of perioperative corneal abrasions in rabbits, it was found that introduction of animals to 3-h general anesthesia resulted in clinically significant chronic damage to the cornea in 50% of cases. The development of the complication was not associated with irreversible or long-term impairment of tear secretion, but it was accompanied by a decrease in tear film stability and growth of the total protein content as well as decrease in total antioxidant activity of the tear induced by low molecular weight antioxidants. In addition, anesthesia-induced changes in activity of tear antioxidant enzymes including superoxide dismutase and enzymes providing homeostasis of reduced glutathione (glutathione peroxidase, glutathione-S-transferase, glutathione reductase) were observed. All these alterations were protracted (up to 1-2 weeks) and therefore might account for transition of the perioperative DES into the chronic form. These findings can be useful in the development of novel approaches for the prevention and treatment of chronic forms of DES in the postanesthetic period.
Keywords: general anesthesia; dry eye syndrome; corneal abrasion; tear secretion; tear film stability; tear proteins; tear antioxidant activity; superoxide dismutase; glutathione peroxidase; glutathione-S-transferase; glutathione reductase

Aging in birds by D. Y. Travin; B. A. Feniouk (1558-1563).
Rodents are the most commonly used model organisms in studies of aging in vertebrates. However, there are species that may suit this role much better. Most birds (Aves), having higher rate of metabolism, live two-to-three times longer than mammals of the same size. This mini-review briefly covers several evolutionary, ecological, and physiological aspects that may contribute to the phenomenon of birds’ longevity. The role of different molecular mechanisms known to take part in the process of aging according to various existing theories, e.g. telomere shortening, protection against reactive oxygen species, and formation of advanced glycation end-products is discussed. We also address some features of birds’ aging that make this group unique and perspective model organisms in longevity studies.
Keywords: aging; senescence; ROS; reactive oxygen species; birds; avian; AGE-products; longevity; telomeres

Role of reactive oxygen species in mast cell degranulation by M. A. Chelombitko; A. V. Fedorov; O. P. Ilyinskaya; R. A. Zinovkin; B. V. Chernyak (1564-1577).
Mast cells are a heterogeneous multifunctional cellular population that promotes connective tissue homeostasis by slow release of biologically active substances, affecting primarily the permeability of vessels and vascular tone, maintenance of electrolyte and water balance, and composition of the extracellular matrix. Along with this, they can rapidly release inflammatory mediators and chemotactic factors that ensure the mobilization of effector innate immune cells to fight against a variety of pathogens. Furthermore, they play a key role in initiation of allergic reactions. Aggregation of high affinity receptors to IgE (FcεRI) results in rapid degranulation and release of inflammatory mediators. It is known that reactive oxygen species (ROS) participate in intracellular signaling and, in particular, stimulate production of several proinflammatory cytokines that regulate the innate immune response. In this review, we focus on known molecular mechanisms of FcεRI-dependent activation of mast cells and discuss the role of ROS in the regulation of this pathway.
Keywords: mast cells; degranulation; reactive oxygen species; FcεRI-signaling