When a theory of aging ages badly

According to the widely acknowledged mitochondrial free radical theory of aging (MFRTA), the macromolecular damage that results from the production of toxic reactive oxygen species (ROS) during cellular respiration is the cause of aging. However, although it is clear that oxidative damage increases during aging, the fundamental question regarding whether mitochondrial oxidative stress is in any way causal to the aging process remains unresolved. An increasing number of studies on long-lived vertebrate species, mutants and transgenic animals have seriously challenged the pervasive MFRTA. Here, we describe some of these new results, including those pertaining to the phenotype of the long-lived Mclk1 ^+/− mice, which appear irreconcilable with the MFRTA. Thus, we believe that it is reasonable to now consider the MFRTA as refuted and that it is time to use the insight gained by many years of testing this theory to develop new views as to the physiological causes of aging.     

Chronic overcrowding decreases cytoplasmic free calcium levels in T lymphocytes of aged CBA/CA mice

Intracellular calcium concentration is a sensitive marker of the homeostasis of living cells, and its increase is an essential step of T lymphocyte activation. Changes in the environment provoke an adaptive stress-response of the organism. In our present work we have investigated the effect of chronic overcrowding on resting and lectin-stimulated cytoplasmic free calcium concentration of splenic T lymphocytes from young and aged CBA/CA mice (50 animals total). The animals were kept under ‘normal’ (68 cm²/animal) or ‘overcrowded’ (22 cm²/animal) conditions for 3 months. Young animals showed no change in resting and stimulated calcium after overcrowding. T cells from aged mice, however, displayed significantly smaller levels of both resting and lectin-stimulated intracellular calcium concentration (p<0.01 each), as compared to those of the non-stressed, aged animals. This inadequate adaptation in the calcium metabolism of T lymphocytes may significantly contribute to the diminished immune response of the aged in stress.     

Genotype-specific reduction in methyl nitrosourea (MNU) induced sister chromatid exchanges (SCE) in vivo during aging

Summary We studied mice from five strains (BALB/c, C3H/HeSnJ, C57BL/6J, Cs^b and 129/ReJ) at two ages (young, 10±1 weeks; and old, 67±3 weeks) for the induction of sister chromatid exchanges (SCEs) in vivo by methyl nitrosourea (MNU). The SCE frequency is genotype-specific. The F₁ phenotype resembles the low responding parent. SCE induction is significantly lower in the older animals of each strain than their younger counterparts, and the reduction of SCE/cell with old age is strain-specific. A general explanation for these results must include strain differences in relative mutagenic sensitivity, genotype-specific pattern of reduction in DNA repair and other such factors affecting SCE formation, with old age.     

A reappraisal of the hormonal regulation of larval fat body histolysis in femaleDrosophila melanogaster

The histolysis of larval fat body cells in adult femaleDrosophila melanogaster was examined in wild type and mutant animals. The fat body cells of wild type (Canton-S),apterous ^56f homozygotes,apterous ^78jts homozygotes and heterozygotes,apterous ⁴/+, ecdysoneless¹ homozygotes and heterozygotes all underwent histolysis normally during the 72 h following adult eclosion. Only in the case ofap ⁴/ap⁴ adults did the cells fail to histolyze normally. The fat body cells of both diapausing and non-diapausing wild type females underwent histolysis at the same rate. Attempts to demonstrate histolysis in vitro were unsuccessful, even in the presence of juvenile hormones (JHs), larval ring glands, or adult ovaries. In all strains other than theap ⁴ homozygotes, a significant proportion of larval fat body cells were dead at any time while theap ⁴/ap⁴ animals, almost all cells remained viable. It is postulated that fat body cell lysis following eclosion is not a JH-mediated event, but is elicited by an as yet unidentified factor(s), possibly originating in the ovary.     

Tissue protection against oxidative stress

We used an enhanced luminescence technique to study the response of rat tissues, such as liver, heart, muscle and blood, to oxidative stress and to determine their antioxidant capacity. As previously found for liver homogenate, the intensity of light emission (E) of tissue homogenates and blood samples, stressed with sodium perborate, is dependent on concentration, and the dose-response curves can be described by the equation E=a·C/exp(b·C). Theb value depends on the antioxidant defence capability of the tissues. In fact, it increases when homogenates are supplemented with an antioxidant, and is correlated with tissue antioxidant capacity, evaluated by two previously set up methods both using the same luminescence technique. Our results indicate that the order of antioxidant capacity of the tissues is liver>blood>heart>muscle. Thea value depends on the systems catalysing the production of radical species. In fact, it is related to the tissue level of hemoproteins, which are known to act as catalysts in radical production from hydroperoxides. The equation proposed to describe the dose-response relation is simple to handle and permits an immediate connection with the two characteristics of the systems analysed which determine their response to the pro-oxidant treatment. However, the equation which best describes the above relation for all the tissues is the following: E=·C/exp(·C). The parameter assumes values smaller than 1, which seem to depend on relative amounts of tissue hemoproteins and antioxidants. The extension of the analysis to mitochondria shows that they respond to oxidative stress in a way analogous to the tissues, and that the adherence of the dose-response curve to the course predicted from the equation E=a·C/exp(b·C) is again dependent on hemoprotein content.     

Effect of estradiol on aminoacid incorporation into proteins of different hypothalamic areas in prepuberal rats

Summary Estradiol in vitro produces a significant increase in the incorporation of³H-leucine into proteins of the anterior hypothalamic area in prepuberal female rats, 15 and 20 days old, but not in younger animals. The ovarian hormone induced no changes in the protein synthetic activity of middle and posterior hypothalamus and cerebral cortex in prepuberal female rats of different ages. Estradiol did not modify the protein synthesis of the hypothalamus and cerebral cortex in prepuberal male rats.Supported by a grant from the Consejo Nacional de Investigaciones Cientifícas y Técnicas de la República Argentina.     

13C NMR-study of osmoregulatory metabolites in the marine molluscTapes watlingi

Summary Examination of homogenates of tissue from the marine bivalve molluscTapes watlingi by natural-abundance¹³C NMR indicates that taurine, betaine, and some free amino acids play a significant role in osmoregulation.Acknowledgments. We thank Dr L.J. Borowitzka and D.D. Jamieson of this Institute for helpful discussions, and the RRIMP marine biology staff for assistance in collecting the animals.Queen Elizabeth II Fellow in Marine Science, 1976–1978. Please address correspondence to this author.     

Increased ALA dehydratase activity and spleen weight in lead-intoxicated rats. A consequence of increased blood cell destruction

Summary Lead was given in the diet (1%) to rats from birth and at different times the animals were studied for delta amino levulinic acid dehydratase (ALAD) activity, spleen weight,⁵⁹Fe incorporation in erythrocytes and⁵¹Cr-labeled erythrocytes survival. The increased ALAD and spleen weight found after lead treatment is explained as a consequence of a shortened survival, which results in a younger age of circulating erythrocytes with higher ALAD activity.Supported by Contract No. 140-76-12 ENV. B. of the E. C. Environmental Research Program.     

Variance to mean ratio and the spatial distribution of animals

Summary Biological and statistical aspects of the application of variance to mean ratio to describe spatial distribution of animals are discussed. It is shown that the parameterb inTaylor's power laws ²=a m ^b shows intra-specific variation depending on the distribution of the constituent units of the population.a andb are only parameters of a very empirical way of describing the relation between variance and mean, which itself is an indicator for spatial distribution. Hence,a andb depend on the distribution behaviour of the animals, and not vice versa.Acknowledgments: The work was developed whilst I was at the Department of Entomology, University of Nairobi. I am grateful to ProfessorT. R. Odhiambo for his interest. I thank the referee for comments.     

An experimental approach to pH measurement in the intercellular free space of higher plant tissues

Summary An experimental approach to the measurement of pH in the free space of plant tissues was made, using both metal minielectrodes and H⁺-sensitive resin glass microelectrodes inserted in small environments artificially created inside the tissue. Experiments were carried out on the parenchyma of higher plants (corn, potato, squash,Senecio herraianus), monitoring pH changes in the free space as a function of the pH of the external medium. The pH in the free space has been found to be different from that of the surrounding solution and its variation in time much slower than would be predicted by assuming that simple diffusion of H⁺ ions took place. The results agree with the hypothesis that the free space behaves as a compartment distinct from both the cytosol and the surrounding medium.     

Effect of NPV of the armywormMythimna (Pseudaletia) separata on the silkwormBombyx mori

Summary Nuclear polyhedrosis virus (NPV) material of the armywormMythimna (Pseudaletia) separata was administered in the form of PIBs and free virus rods to the larvae of silkwormBombyx mori. Routes used for administration were topical, intrahaemocoelic and oral. The larvae were treated with following concentrations: 10×10⁵ PIBs/L, 10×10⁶ PIBs/L, 10×10⁷ PIBs/L, 10×10⁸ PIBs/L. In all the 3 experiments, the larvae showed neither any signs and symptoms, nor mortality due to polyhedrosis. Thus it appears that the NPV ofM. (P.) separata is safe for the silkwormB. mori.Acknowledgments. We are grateful to the Karnatak University, Dharwad, for awarding UGC Junior Fellowship to S.G.D. and for financial assistance made available to S.B.M. We thank Professor M. Appaswamy Rao, Department of Zoology, for providing the necessary facilities. Our thanks are also due to Mr B. Chavannawar for supplying mulberry leaves and to the Government Silk Farm, Hindalaga, Belgaum, for providing silkworm eggs.     

Structural characteristics and distribution of satellite cells along crayfish muscle fibers

The distribution of satellite cells (sc) in long-sarcomere muscle fibers from the carpopod extensor muscle of the crayfish (Astacus fluviatilis) has been studied electron-microscopically. The sc are spindle-shaped and are oriented parallel to the long axis of a fiber. The mean lengths of sc nuclei (17.00 m) and that of myonuclei (18.35 m) differ non-significantly. In older animals, the mean ratio of the number of sc nuclei to the total number of nuclei (sc nuclei + myonrclei) is 0.0716, 0.0848, and 0.034 for the tendon, central and shell segments, respectively. The corresponding values for younger animals are 0.158, 0.166, and 0.081. The mean numbers of sc nuclei per mm of a fiber are 94, 117, and 47 (older animals), and 164, 117, and 94 (younger animals) for the tendon, central and shell segments, respectively. The high incidence of sc per unit fiber length in crayfish may be related to the fact that crayfish muscle fibers have a much larger diameter than vertebrate muscle cells.     

Did Einstein prove E=mc2?

Although Einstein's name is closely linked with the celebrated relation E=mc² between mass and energy, a critical examination of the more than half dozen “proofs” of this relation that Einstein produced over a span of forty years reveals that all these proofs suffer from mistakes. Einstein introduced unjustified assumptions, committed fatal errors in logic, or adopted low-speed, restrictive approximations. He never succeeded in producing a valid general proof applicable to a realistic system with arbitrarily large internal speeds. The first such general proof was produced by Max Laue in 1911 (for “closed” systems with a time-independent energy–momentum tensor) and it was generalized by Felix Klein in 1918 (for arbitrary time-dependent “closed” systems).     

High affinity recognition of a <Emphasis Type="Italic">Phytophthora</Emphasis> protein by <Emphasis Type="Italic">Arabidopsis</Emphasis> via an RGD motif

The RGD tripeptide sequence, a cell adhesion motif present in several extracellular matrix proteins of mammalians, is involved in numerous plant processes. In plant-pathogen interactions, the RGD motif is believed to reduce plant defence responses by disrupting adhesions between the cell wall and plasma membrane. Photoaffinity cross-linking of [¹²⁵I]-azido-RGD heptapeptide in the presence of purified plasma membrane vesicles of Arabidopsis thaliana led to label incorporation into a single protein with an apparent molecular mass of 80 kDa. Incorporation could be prevented by excess RGD peptides, but also by the IPI-O protein, an RGD-containing protein secreted by the oomycete plant pathogen Phytophthora infestans. Hydrophobic cluster analysis revealed that the RGD motif of IPI-O (positions 53–56) is readily accessible for interactions. Single amino acid mutations in the RGD motif in IPI-O (of Asp⁵⁶ into Glu or Ala) resulted in the loss of protection of the 80-kDa protein from labelling. Thus, the interaction between the two proteins is mediated through RGD recognition and the 80-kDa RGD-binding protein has the characteristics of a receptor for IPI-O. The IPI-O protein also disrupted cell wall-plasma membrane adhesions in plasmolysed A. thaliana cells, whereas IPI-O proteins mutated in the RGD motif (D56A and D56E) did not.Received 23 October 2003; received after revision 5 December 2003; accepted 12 December 2003     

Aberrant,canavanyl protein formation and the ability to tolerate or utilize L-canavanine

Summary L-Canavanine, 2-amino-4-(guanidinooxy)butyric acid, and L-arginine incorporation into de novo synthesized proteins was compared in six organisms. Utilizing L-[guanidinooxy¹⁴C]canavanine and L-[guanidino¹⁴C]arginine at substrate saturation, the canavanine to arginine incorporation ratio was determined in de, novo synthesized proteins.Caryedes brasiliensis andSternechus tuberculatus, canavanine utilizing insects;Canavalia ensiformis, a canavanine storing plant; and to a lesser extentHeliothis virescens, a canavanine resistant insect, failed to accumulate significant canavanyl proteins. By contrast,Manduca sexta, a canavanine-sensitive insect, andGlycine max, a canavanine free plant, readily incorporated canavanine into newly synthesized proteins. This study supports the contention that the incorporation of canavanine into proteins in place of arginine contributes significantly to canavanine's antimetabolic properties.     

The effect of chronic heat stress on cortisol levels in the antarctic fishPagothenia borchgrevinki

Radioimmunoassay was used to determine levels of the stress-inducible glucocorticoid, cortisol, circulating in the plasma of the extremely stenothermal Antarctic fishPagothenia borchgrevinki at rest and after heat stress. Fish sampled immediately after capture (–1.9°C) had low cortisol levels (10.4±1.4 ng ml^–1, mean±SEM) as did fish which were laboratory rested for 3 days. Sudden exposure to 5°C (48h) resulted in a peak cortisol value after 3 h (69.9±6.8 ng ml^–1) whereas exposure to 8°C (6h) resulted in a peak value after 1 h (73.5±8.0 ng ml^–1). At both temperatures levels remained significantly elevated (p<0.05) for the entire period of exposure. Increased temperature also resulted in a significant change in haemoglobin, haematocrit and mean cell haemoglobin concentration (MCHC) (p<0.05). Plasma lactate was significantly elevated only after exposure to 8°C (p<0.05). Plasma cortisol levels fromP. borchgrevinki are reported here for the first time and show this cryopelagic Antarctic species to have an unusual hormonal stress profile.     

Abbauprodukte von Simazin in Gramineen

Summary Coix lacryma-jobi L., about 20 days old and grown on watercultures, were fed with¹⁴C-ring-labelled simazine. After freeze drying, the plant tissue was extracted with chloroform and 80% ethanol. The ethanol extractable activities were fractionated on a cation exchange column. The radioactive metabolites seem to be identical with 2-hydroxy-4, 6-diethylamino-s-triazine (hydroxysimazine), 2-hydroxy-4, 6-diamino-s-triazine (ammelin) and 2-hydroxy-4-ethylamino-6-amino-s-triazine.     

Thermodynamics of bromate and iodate of sodium in dioxane-water mixtures at different temperatures

Summary The free energy transfer (G _t ⁰ ) enthalpy of transfer (H _t ⁰ ) and entropy of transfer (S _t ⁰ ) of NaBrO₃ and NaIO₃ from water to 10, 20 and 30% dioxane-water mixtures have been studied using conductance measurements. The chemical and electrical parts of these transfers of thermodynamic parameters have been estimated.To whom reprint requests should be addressed.     

Effect of various stressors on the levels of lipid peroxide,antioxidants and Na+, K+-ATPase actrivity in rat brain

The level of malondialdehyde (MDA), an index of lipid peroxidation, and the antioxidants superoxide dismutase (SOD) and glutathione (GSH), as well as the activity of Na⁺, K⁺-ATPase, were assessed in whole rat brain after immobilization, anemic hypoxia (NaNO₂) and 72 h starvation. The effect of these stressors on plasma glucose and corticosterone levels was also observed. Hypoxia and starvation stimulated the lipidj peroxide formation in braini as indicated by an increase in the level of MDA, being higher after starvation than hypoxia. Brain SOD activity was also increased in response to hypoxia and starvation while GSH content was only diminished ini hypoxia. However, neither MDA nor antioxidants were affected by immobilization. On the other hand, the activity of brain Na⁺, K⁺-ATPase was significantly increased by immobilization and hypoxia but decreased in starvation. A similar pattern of change was also observed in plasma glucose and corticosterone levels in response to these stressors. These results elucidate differences in the biochemical response of animals towards various types of stress, with increased lipid peroxide formation in hypoxia and starvation.     

Pauling's Defence of Bent-Equivalent Bonds: A View of Evolving Explanatory Demands in Modern Chemistry

Linus Pauling played a key role in creating valence-bond theory, one of two competing theories of the chemical bond that appeared in the first half of the 20th century. While the chemical community preferred his theory over molecular-orbital theory for a number of years, valence-bond theory began to fall into disuse during the 1950s. This shift in the chemical community's perception of Pauling's theory motivated Pauling to defend the theory, and he did so in a peculiar way. Rather than publishing a defence of the full theory in leading journals of the day, Pauling published a defence of a particular model of the double bond predicted by the theory in a revised edition of his famous textbook, The Nature of the Chemical Bond. This paper explores that peculiar choice by considering both the circumstances that brought about the defence and the mathematical apparatus Pauling employed, using new discoveries from the Ava Helen and Linus Pauling Papers archive.