A correlation between shortened life span and UV-sensitivity in some strains of Saccharomyces cerevisiae.

Summary From a UV-irradiated sample of diploid cells several clones were isolated, which produced cells with a shortened life span. A closer examination of three of these clones showed among other deviations from the wild type a higher sensitivity to UV-irradiation. Three other clones, which were selected from a haploid strain as UV-sensitive mutants, proved to be shortlived as well. In all these strains photoreactivation and liquid holding reactivation were unimpaired. There was no cross-sensitivity to X-irradiation. The correlation between shortened life span and UV-sensitivity is discussed.These studies were aided in part by a grant of the Deutsche Forschungsgemeinschaft.     

Mammalian growth and development parameters, cell proliferation in culture and the maximal life span]

The maximum life span of mammals is known to be proportional to the pregnancy duration and to the age at puberty. We found that the maximum life span of mammals was also proportional to the number of cell doublings, and inversely proportional to the rate of duplication of these cells, during embryogenesis or for the time from zygote formation to growth termination. We found also that the life span of "stationary phase aging" transformed Chinese hamster cells (time from subcultivation until culture "death", i.e., until the moment when the number of live cells is less than 10% of their number at saturation density) was proportional to the duration of their growth and number of cell doublings during the period from subcultivation to saturation density, and inversely proportional to the rate of cell culture duplication during the same period. The dependencies for cell cultures and mammals proved to be analogous to each other. An approximately twofold decrease in the cell duplication rate, as a result of a decrease of the growth medium temperature from 37 to 27 degrees C or the introduction of ethanol to a final concentration 2%, increased the life span of "stationary phase aging" cultures more than twofold. The data obtained suggest that influences resulting in optimized delay of the rate of cell duplication, and correspondingly the mean rate of proliferation during the period of growth in mammals, may increase their maximum life span.     

An investigation of effects of toluene and cigarette smoking on some blood parameters and lymphocyte life span

As toluene is an organic solvent, its cytotoxic effect on the cell is known. Similarly, it has been demonstrated that many of the chemical agents that enter the body through smoking have cytotoxic and genotoxic effects on the cells. In this study, the effects of these two toxic agents, both separately and in combination, on leukocyte counts, lymphocyte counts and mitotic index values were investigated. The study was carried out on blood samples of 100 males, divided into four groups: 25 non-smokers and 25 smokers, 25 toluene-exposed non-smokers and 25 no toluene-exposed smokers. The blood cell values of the blood samples were determined automatically on the hemogram apparatus. In addition slides of the blood samples were prepared according to the chromosome analysis procedure and the mitotic index values were determined through microscopy. The possible effects of smoking and toluene on lymphocyte life span was considered by correlating mitotic index values with lymphocyte counts in the same way for each of the subgroups. Results revealed that leukocyte counts and mitotic index values were higher in the smokers than the non-smokers whether or not they had been exposed to toluene. In addition the results indicate that lymphocyte life span may be shortened due to cigarette smoking and toluene exposure.     

Oxidative damage to mitochondrial DNA is inversely related to maximum life span in the heart and brain of mammals.

DNA damage is considered of paramount importance in aging. Among causes of this damage, free radical attack, particularly from mitochondrial origin, is receiving special attention. If oxidative damage to DNA is involved in aging, long-lived animals (which age slowly) should show lower levels of markers of this kind of damage than short-lived ones. However, this possibility has not heretofore been investigated. In this study, steady-state levels of 8-oxo-7, 8-dihydro-2'-deoxyguanosine (8-oxodG) referred to deoxyguanosine (dG) were measured by high performance liquid chromatography (HPLC) in the mitochondrial (mtDNA) and nuclear (nDNA) DNA from the heart of eight and the brain of six mammalian species ranging in maximum life span (MLSP) from 3.5 to 46 years. Exactly the same digestion of DNA to deoxynucleosides and HPLC protocols was used for mtDNA and nDNA. Significantly higher (three- to ninefold) 8-oxodG/dG values were found in mtDNA than in nDNA in all the species studied in both tissues. 8-oxodG/dG in nDNA did not correlate with MLSP across species either in the heart (r=-0.68; P<0.06) or brain (r = 0.53; P<0.27). However, 8-oxodG/dG in mtDNA was inversely correlated with MLSP both in heart (r=-0.92; P<0.001) and brain (r=-0.88; P<0.016) tissues following the power function y = a(.)x(b), where y is 8-oxodG/dG and x is the MLSP. This agrees with the consistent observation that mitochondrial free radical generation is also lower in long-lived than in short-lived species. The results obtained agree with the notion that oxygen radicals of mitochondrial origin oxidatively damage mtDNA in a way related to the aging rate of each species.-Barja, G., Herrero, A. Oxidative damage to mitochondrial DNA is inversely related to maximum life span in the heart and brain of mammals.     

The linear allometric relationship between total metabolic energy per life span and body mass of mammals

The aim of this study is to establish and calculate the exact allometric relationship between the total metabolic energy per life span and the body mass in a wide range of mammals with about six orders of magnitude variation of the body mass of animals. The study shows that it exists a linear relationship between the total metabolic energy per life span PT(ls) (kJ) and the body mass M (kg) of 95 mammals (3 monotremes, Subclass Prototheria, 16 marsupialis (Subclass Theria, Infraclass Metatheria) and 76 placentals (Subclass Theria, Infraclass Eutheria)) from type: PT(ls)=A(ls)(+)M(1.0511), where P (kJ/day) is the basal rate of metabolism and T(ls) (days) is the mean life span of animals. The linear coefficient A(ls)(+)=7.158x10(5) kJ/kg is the total metabolic energy, exhausted during the life span per 1 kg body mass of the animals. The mean values of the total metabolic energy per life span, per unit body mass (A(ls)) for orders from Subclass Prototheria and Theria (Infraclass Metatheria) and orders Xenarthra, Pholidota, Soricomorpha, Rodentia (Infraclass Eutheria) varied negligible in interval (4.656-5.80)x10(5) kJ/kg. The coefficient A(ls) grows from (7.68-8.36)x10(5) kJ/kg in Lagomorpha and Artiodactyla (Eutheria) to (10.58-12.64)x10(5) kJ/kg in orders Carnivora, Pinnipeda and Chiroptera (Eutheria). A(ls) grows maximum to 18.5x10(5) kJ/kg in Primates. Thus, the values of coefficient A(ls) differ maximum four-fold in all orders. Across the all species the values of A(ls) are changes about one order of magnitude. Consequently, our survey shows that the changes of the body mass, basal metabolic rate and the life span of animals are three mutually related parameters, so that the product A(ls)=(PT(ls))/M remains relatively constant in comparison to 1 million fold difference in body mass and total metabolic energy per life span between mammals.     

Methylcellulose effect on cell proliferation and glucose utilization in chemically defined medium in large stationary cultures

Three different established strains of mammalian cells were grown in chemically defined medium in large cultures. The degree of proliferation of cells of an established strain from human skin in large stationary cultures was significantly greater in the presence of methylcellulose (medium NCTC 135M) than in its absence (medium NCTC 135). The relatively fragile cells of a derivative of monkey kidney LLCMK2 strain were carried in large stationary cultures through 11 transfer generations during 152 days. The presence of methylcellulose was associated with higher cell population levels, proliferation rates, and cell viability. Cells of this strain utilized glucose at an extremely high rate; during two representative periods the rate averaged 1.2 mg/10⁶ cells/day in cultures on medium 135M and 1.9 mg in medium 135. In a 53-day experiment with mouse fibroblast 2071-L cells, the cells in suspension culture during the first 28 days went through the normal lag, logarithmic plateau, and initial decline phases in medium 135M, and then were transferred to large stationary cultures, where they proliferated for 7 days at uniformly high rates in both medium 135 and medium 135M. It appeared that cells of strain 2071-L in such stationary cultures had no need for Methocel as a protective agent. Glucose utilization rates while these cells were carried in large stationary cultures averaged 2–4 times the rates while they were in suspension cultures: about 0.8 and 0.2 mg/10⁶ cells/day, repectively.     

Influence of environmental factors in early ontogenesis on aging and life span

A possible dependence of aging and life span in man on specific features of early ontogenesis has been analyzed on the basis of published and author's data. It was shown that the life span depended on climatic factors acting during prenatal and/or early postnatal period. When analyzing a sample of 101,634 humans died in Kiev in 1990-2000, a reliable relationship was established between the age of death and month of birth. The lowest and highest death ages were observed in persons born in April-June and in the end of the year, respectively. The lowest and highest average monthly values of death age differed in men and women by 2.6 and 2.3 years, respectively. In persons survived for more than 60 years, a "birthday" effect was demonstrated: dependence of the mortality level on the month of individual annual cycle was the highest in its first and last month. It has been proposed that this effect is related imprinting of the "birth stress" in the structure of biological rhythms, which may lead to periodic changes in viability during the individual annual cycle. The mechanisms underlying long-term effects of stresses in early ontogenesis have been studied on Drosophila melanogaster. X-irradiation of eggs at 0.50 and 0.75 Gy led to an increased level of survival of imago. The described radiation hormesis was accompanied by an increased resistance of DNA to S1-nuclease, which could be due to a long-term activation of the repair system in irradiated insects. The results obtained suggest that long-term changes in the range of gene expression induced by mild stresses in early ontogenesis can be a key mechanism of early "programming" of the potential of longevity and hormesis for life span.     

Some remarks on the relationship between autophagy,cell aging,and cell proliferation restriction

In the review, the main types of autophagy (macroautophagy, microautophagy, and chaperonemediated autophagy) are shortly described. Data about the character of the influence of autophagy on the aging process and on the development of some neurodegenerative diseases in various organisms are analyzed. It is noted that this effect is usually (though not always) beneficial. Results of investigations of the phenomenon in experiments on mice, nematodes, fruit flies, bacteria, yeast, and cell cultures of higher organisms are considered. Obvious relationship between autophagy activation and cell proliferation restriction is emphasized. The latter, in our opinion, is the main cause of age-related accumulation of various defects (the most important of them is DNA damage) in cells and tissues, which leads to an increase in the death probability (i.e., to aging). It is concluded that studies of the role of autophagy in the aging process on the models of chronological aging in yeast or stationary phase aging of cell cultures could be considered as the most appropriate approach to the problem solution.     

A comparison of the proliferative and replicative life span kinetics of cell cultures derived from monozygotic twins

Summary We have compared the growth rates, kinetics of cell aging, and replicative life spans of skin fibroblast cell cultures derived from three pairs of monozygotic twins of similar ages. The results of these studies indicated no significant differences in the cell densities 7 days after inoculation or replicative life spans within each twin pair but highly significant differences among each twin pair. The kinetics by which each culture aged ([³H]thymidine-labeled nuclei) were compared within and among the twin cell cultures. Although the slopes of each regression line were not significantly different, comparisons of the elevations of each line supported the conclusion that the aging of monozygotic twin cell cultures is similar within the twin pairs but differs among the twin pairs. This research was supported by IIT Research Institute.     

Inverse correlation between species life span and capacity of cultured fibroblasts to metabolize polycyclic hydrocarbon carcinogens.

Many investigators have hypothesized that the aging process may result from an accumulation of DNA damage, and, if valid, this necessitates a means by which this accumulation can be related to the potential life span of an organism. Using an assay for cell-mediated mutagenesis, we have tested multiple diploid fibroblast strains from six mammalian species of widely differing life spans, and found a very good inverse correlation between species life span and ability to activate 7,12-dimethylbenz(a)anthracene (DMBA) to mutagenic forms. We have also found a very good inverse correlation between species life span and ability to activate DMBA to forms capable of covalent binding to DNA. Since the polycyclic hydrocarbon carcinogens such as DMBA and benzo(a)pyrene (BP) are chemically non-reactive in their native forms and must be metabolically activated by mixed-function oxidases to their biologically active forms, these data indicate that the capacity of fibroblasts to activate polycyclic hydrocarbon carcinogens to DNA-damaging forms is a species property related to potential life span. To determine the role of carcinogen metabolism in this phenomenon the capacity of diploid fibroblasts from eight mammalian species to convert BP and DMBA to water-soluble metabolites was then determined. This rate of conversion varies widely among different species and shows a very good inverse correlation with species life span. As a whole, these findings suggest that the ability of cultured cells to metabolize the polycyclic hydrocarbon carcinogens is related to species life span, and may be important in the occurrence of spontaneous cancer.     

The 'transition probability model' and the regulation of proliferation of human diploid cell cultures during aging.

Genealogies derived from time-lapse cinemicrophotographic studies of aging human diploid cell cultures were analysed in terms of the 'transition probability' model. It was found that the distribution of intermitotic times obtained from middle passage cells deviated only slightly from that predicted by the model. In contrast, the plot for late passage cultures did not fit the predicted pattern and appeared to be composed of multiple curves. These changes are discussed in reference to cellular senescence as expressed by normal human diploid cells in vitro.     

The nature of and the interrelations between the concepts of "life","disease" and "aging"

An attempt is made to consider disease and aging following from the concepts of the essence of life. The proposed definition of life represents a modified Engels' (1878) definition. Proceeding from the analysis of possible mechanisms of different disturbances in the life process leading to a decreased probability of the organism existence it is concluded that disease develops either as a result of hereditary changes in the genome or due-to disorders in its realization under certain unfavorable conditions. Aging is determined by the properties of the genome itself and develops in connection with age increase.     

The effect of UV irradiation on proliferation and life span of human diploid fibroblast-like cells

Summary The effect of low dose UV irradiation on the reinitiation of proliferative activity and on the life span of human diploid fibroblast-like cells is described. Cells were exposed to UV at confluence or after maintenance in an arrested state. Cell division was stimulated immediately after UV irradiation or after an additional post-UV incubation period. Arrested populations of all in vitro ages exhibited a greater sensitivity to UV and the reinitiation of proliferation was enhanced by post-UV incubation before stimulation. Ultraviolet light had no effect on life span regardless of in vitro cell age, culture state at the time of exposure, or the presence of a postirradiation period of arrest.     

Effects of establishing cell cultures and cell culture conditions on the proliferative life span of human fibroblasts isolated from different tissues and donors of different ages

It is well known that normal human cells placed in a culture environment exhibit a limited proliferative capacity. The extent to which the culture environment influences proliferative life span is not understood. This study evaluated the effects of the standard procedures used to establish and maintain cultures on the proliferative life spans of different types of human fibroblast cells established from fetal and adult skin and lung. The results of this study demonstrate that procedures to establish cell cultures use only one of several subpopulations of cells present in biopsy pieces and that the culture conditions routinely employed by most laboratories can exert significant effects on proliferative life-span determinations. The maximum proliferative life span differed significantly when obtained by growing the cells in two commonly used commercial media. Proliferative life span was inversely related to ambient oxygen tension and directly related to seeding density in all of the lines examined although lines established from adult skin were much more resistant to toxicity. Enzymatic antioxidant defense levels of fetal skin fibroblasts were much lower than those observed in adult skin fibroblasts, but the effects of oxygen on their life spans were similar. Hyperoxia induced larger increases in glutathione concentration in cell lines with low antioxidant enzyme levels.     

Effect of long-term somatotropin treatment on body composition and life span in aging obese Zucker rats

The objective of this work was to test the hypothesis that a somatotropin (STH)-induced reduction in body fat would prolong the life span of the obese Zucker rat. Two experiments were conducted. In the first experiment, male and female, lean and obese Zucker rats were treated with STH (0 or 2 mg/d bovine STH) for 4 weeks, beginning at 7 months of age. Across phenotypes, STH treatment increased the growth rate by 159%, muscle weights by 14%, and circulating insulin-like growth factor (IGF)-1 by 23%, and decreased carcass fat by 21% (P < 0.05). The second experiment was a longevity trial to determine whether these changes in body composition would increase the life span of the obese rat. Beginning at 7 months of age, individually housed, male and female, lean and obese rats were assigned to daily STH treatments (0 or 2 mg/d). Rats were monitored daily, and sick or moribund rats were euthanized and necropsied to determine existing pathologies. The average life span of the lean rats was 661 days and was unaffected by STH treatment (639 days, NS) or gender. Average life span of the vehicle-injected obese rats (435 days) was less than that of the lean group (P < 0.001). STH treatment of the obese rats resulted in a further reduction of life span (349 days, P < 0.02). The predominant pathology observed across the treatment groups was renal disease, characterized by progressive glomerulonephropathy. Thus, although exogenous STH was able to reduce carcass lipid and to increase lean tissue mass in obese rats, there was no improvement in longevity. In contrast to the hypothesis, STH actually reduced the life span of the obese rat. It is likely that STH treatment accelerated the development of progressive glomerulonephropathy in the obese rat.