Fine-structural changes in the midgut of old Drosophila melanogaster.

Variations of the mitochondrial structure with increasing age are of particular interest because the production of ATP is necessary for most cellular activities. Therefore, a senescent decrease of mitochondrial activity could jeopardize cell survival and function. We combined a statistical with a morphological approach to investigate these changes in mitochondrial size and structure of Drosophila melanogaster. The midgut epithelium is of special interest because previous research shows striking aging changes. Investigations of the mitochondria of flies of three age groups (10, 39, and 65 days) reveal an increase in size as well as in its standard deviation. This is linked to the fact that mitochondrial enlargement occurs especially in the luminal region. Ultrastructural studies show that with increasing age the mitochondria become more osmiophilic, their cristae disarranged, cristae-free areas can also be found, and there is an accumulation of homogeneous, granular and lamellar dense bodies. In addition, an accumulation of virus-like particles, fiber-shaped structures, reticular structures, and dense bodies without a surrounding membrane was found.

Accelerated aging of fasted Drosophila. Preservation of physiological function and cellular fine structure by thiazolidine carboxylic acid (TCA).

Thiazolidine carboxylic acid (TCA) is a natural liver metabolite whose Mg-salt increased lifespan of flies and mice (Miquel and Economos, 1979, Exp. Geront. 14: 279). We studied the physiological and cellular fine structural effects of various concentrations of TCA in the food of male Drosophila. Flies on 0.3% TCA at 27 degrees C had a reduced oxygen consumption rate (about 20% less than controls) at 3 wks of age while their mating capacity and speed of mating were preserved; the flies lived in various experiments 20-30% longer than controls. Apparently TCA improved the metabolic efficiency of the flies (possibly from less "waste" of energy due to improved mitochondrial coupling). However, flies on 0.9% TCA had a reduced mating capacity and lifespan (:toxicity) while at 0.1% TCA was ineffective. A similar dose-response relationship was found in young flies treated with TCA for 1 week and then deprived of food and water, a procedure found to induce accelerated physiological aging. TCA at the 0.3% and 0.6% level reduced the speed of development and the size of the enclosed flies. Electron microscopic investigation of wing muscle showed that 0.3% TCA had a protective effect on cellular fine structure. Though in starved controls (40% survivors after 24 hours of starvation) there was a total absence of glycogen granules, and a striking shrinkage and densification of mitochondria, TCA to a large extent protected muscle cells from these effects of starvation.

Modification of a Kowa RC-2 fundus camera for self-photography without the use of mydriatics.

Research on retinal circulation during space flight required the development of a simple technique to provide self monitoring of blood vessel changes in the fundus without the use of mydriatics. A Kowa RC-2 fundus camera was modified for self-photography by the use of a bite plate for positioning and cross hairs for focusing the subject's retina relative to the film plane. Dilation of the pupils without the use of mydriatics was accomplished by dark adaption of the subject. Pictures were obtained without pupil constriction by the use of a high speed strobe light. This method also has applications for clinical medicine.

Effects of simulated increased gravity on the rate of aging of rats: implications for the rate of living theory of aging.

Ever since Pearl proposed the rate of living theory of aging numerous studies have demonstrated its validity in poikilotherms. In mammals, however, satisfactory experimental demonstration is still lacking because an externally imposed increase of basal metabolic rate of these animals (e.g. by placement in the cold) is usually accompanied by general homeostatic disturbance and stress. The present study was based on the finding that rats exposed to slightly increased gravity are able to adapt with little chronic stress but at a higher level of basal metabolic expenditure (increased 'rate of living'). The rate of aging of 17-mth-old rats that had been exposed to 3.14 times normal gravity in an animal centrifuge for 8 mth was larger than of controls as shown by apparently elevated lipofuscin content in heart and kidney, reduced numbers and increased size of mitochondria of heart tissue, and inferior liver mitochondria respiration (reduced 'efficiency': 20% larger ADP: 0 ratio, P less than 0.01; reduced 'speed': 8% lower respiratory control ratio, P less than 0.05). On the other hand, steady-state food intake per day per kg body weight, which is presumably proportional to 'rate of living' or specific basal metabolic expenditure, was about 18% higher than in controls (P less than 0.01) after an initial 2-mth adaptation period. Finally, though half of the centrifuged animals lived only a little shorter than controls (average about 343 vs. 364 days on the centrifuge, difference statistically nonsignificant), the remaining half (longest survivors) lived on the centrifuge an average of 520 days (range 483-572) compared to an average of 574 days (range 502-615) for controls, computed from onset of centrifugation, or 11% shorter (P less than 0.01). Therefore, these results show that a moderate increase of the level of basal metabolism of young adult rats adapted to hypergravity compared to controls in normal gravity is accompanied by a roughly similar increase in the rate of organ aging and reduction of survival, in agreement with Pearl's rate of living theory of aging, previously experimentally demonstrated only in poikilotherms.

Long term effects of low doses of 56Fe ions on the brain and retina of the mouse: ultrastructural and behavioral studies.

Eight month old male C57BL6 mice were exposed without anesthesia to whole-body irradiation in circular holders. The mice were tested for behavioral decrements after 0.5 and 50 rads of Fe particle irradiation at 6 and 12 months post irradiation to obtain long term results. A standard maze was used and the animals were timed for completion thereof. A string test also was administered to the mice, testing their ability to grasp and move along a string to safety. The results from animals exposed to 50 rads were significantly different from [correction of fron] control results to p = < .001 in both systems of testing. The hippocampus (believed to be the location of environmental interaction in the brain) and the retina were examined for ultrastructural changes. The ultrastructural changes were similar to those we found in our Cosmos 782, 936 and in our Argon experiments. The mouse data indicate that iron particles were able to induce long term changes in the central nervous system which lead to behavioral impairment.

Comparative study of spermatogonial survival after x-ray exposure, high LET (HZE) irradiation or spaceflight.

Spermatogonial cell loss has been observed in rats flown on Space Lab 3, Cosmos 1887, Cosmos 2044 and in mice following irradiation with X-ray or with high energy (HZE) particle beams. Spermatogonial loss is determined by cell counting in maturation stage 6 seminiferous [correction of seminferous] tubules. With the exception of Iron, laboratory irradiation experiments (with mice) revealed a similar pattern of spermatogonial loss proportional to the radiation dose at levels less than 0.1 Gy. Helium and Argon irradiation resulted in a 5% loss of spermatogonia after only 0.01 Gy exposure. However, significant spermatogonial loss (45%) occured at this radiation level with Iron particle beams. The loss of spermatogonia during each space flight was less than 10% when compared to control (non-flight) animals. This loss, although small, was significant. Although radiation may be a contributing factor in the loss of spermatogonia during space flight, exposure levels, as determined by dosimetry, were not significant to account for the total cell loss observed.

Ultrastructural changes in tracheal epithelial cells exposed to oxygen.

White albino rats were sacrificed after 24, 36, 48, 72, and 96 h of exposure to 100% O2 at 1 atm. Tissue was prepared for the scanning electron microscope (SEM) by Critical Point Drying and for the transmission electron microscope (TEM) by plastic embedding. Scanning microscopy showed a loss of microvilli after 48 h of exposure. Cilia appeared relatively normal with SEM, but TEM revealed changes in the outer membrane. In TEM, nonciliated cells appeared swollen and often encroached on the ciliated cells. A heavy mucous blanket remained even after processing. All the changes observed that are induced by oxygen exposure contribute to mucostasis, reducing and/or halting mucociliary clearance.

The effects of cosmic particle radiation on pocket mice aboard Apollo XVII: V. preflight studies on tolerancee to oxygen and heat. Part III. effects on eyes.

A study was made of the eyes of eight pocket mice exposed to oxygen at partial pressures of 8, 10, or 12 psi over a period of 7 d. At the termination of the exposure, the animals were decompressed to sea-level O2 either immediately or over a period of 30, 60, or 90 min. No pathological changes were found in any of the eyes, except in the retina of one of the animals exposed to 12 psi O2. Here, only a single rod photoreceptor was found damaged, an observation not regarded as significant. Hence, an oxygen partial pressure as high as 12 psi in the camister in which pocket mice were expected to fly on Apollo XVII would probably have no deleterious effect on the eyes of the animals.

The effects of cosmic particle radiation on pocket mice aboard Apollo XVII: VII. Cosmic ray particle dosimetry and trajectory tracing.

Five pocket mice (Perognathus longimembris) were flown on Apollo XVII, each with a solid-state (plastic) nuclear track detector implanted beneath its scalp. The subscalp detectors were sensitive to HZE cosmic ray particles with a LET larger than or equal to 0.15 million electron volts per micrometer (MeV/mjm). A critical aspect of the dosimetry of the experiment involved tracing individual particle trajectories through each mouse head from particle tracks registered in the individual subscalp detectors, thereby establishing a one-to-one correspondence between a trajectory location in the tissue and the presence or absence of a lesion. The other major aspect was the identification of each registered particle. An average of 16 particles with Z larger than or equal to 6 and 2.2 particles with Z larger than or equal to 20 were found per detector. The track density, 29 tracks/cm2, when adjusted for detection volume, was in agreement with the photographic emulsion data from an area dosimeter located next to the flight package.

The effects of cosmic particle radiation on pocket mice aboard Apollo XVII: XI. Results of eye examination.

Five pocket mice (Perognathus longimembris) were flown on Apollo XVII, and four survived. All the eyes, except one eye from the dead flight mouse, were examined histologically. In the four surviving mice, a total of five cosmic ray particles which had registered in the subscalp particle detectors had trajectories that intersected the eyes. Four of them (Z equal to 6-9 for three of the particles and Z larger than or equal to 10 for the fourth) most likely went through the head before reaching the particle detector, while the thindown direction of the fifth (Z larger than or equal to 10) was not determinable. The retinas of the flight animals were found free from histological alterations such as might have been expected from encounters with cosmic ray particles.

Retinal changes in rats flown on Cosmos 936: A cosmic ray experiment.

Ten rats, five centrifuged during flight to simulate gravity and five stationary in flight and experiencing hypogravity, orbited the Earth. No differences were noted between flight-stationary and flight-centrifuged animals, but changes were seen between these two groups and ground controls. Morphological alterations were observed comparable to those in the experiment flown on Cosmos 782 and to the retinal cells exposed to high-energy particles at Berkeley. Affected cells in the outer nuclear layer showed swelling, clearing of cytoplasm, and disruption of the membranes. Tissue channels were again found, similar to those seen on 782. After space flight, preliminary data indicated an increase in cell size in montages of the nuclear layer of both groups of flight animals. This experiment shows that weightlessness and environmental conditions other than cosmic radiation do not contribute to the observed damage of retinal cells.

Cosmic ray effects on the eyes of rats flown on Cosmos No. 782, experimental K-007.

The eyes from six rats were fixed at the recovery site in Russia after they had circled the earth for 19.5 d in a 62.8 degree orbit. The eyes of six more flight rats were fixed 25 d later. These two preparations and eyes exposed to 1000 rad of neon and argon were compared to obtain data on possible radiation effects on the retina. Most of the flight eye tissue was normal; however, necrotic nuclei were found in the outer nuclear layer and channels were located in the outer segment area. Macrophages were seen between the pigment layer and outer segments. Comparison of the Day zero znd 25-d postflight eyes suggested some possible recovery. Light flashes seen by space travelers and damage from cosmic rays appear to arise from two different sites of interaction. The flashes are created by cosmic ray transversal of the outer segments while pathologic change, when it occurs, is quite possibly from interaction with some part of the nucleus. Nevertheless, direct interaction with other cellular components could also occur. Other factors, such as secondaries from spacecraft shielding, may play an important role.

The effects of cosmic particle radiation on pocket mice aboard Apollo XVII: VI. launch, flight, and recovery.

The final phase to fly five pocket mice in the Apollo XVII command module was carried out at the NASA Kennedy Space Center. Upon completion of the 13-d space flight, the package was removed from the spacecraft and, after having been purged with an oxygen-helium gas mixture, was flown to American Samo. Four of the five mice were recovered alive from the package. Analysis of the mouse that died during the flight revealed several factors that could have contributed to its death, the chief of which was massive hemorrhage in its middle ear cavities.

Effects of high energy particle (HZE) radiation on the distal lung.

Major sources of concern for manned space travel are the effects of high energy particle (HZE) radiation on various biological systems, and the consequences of major solar activity. To date, considerable attention has been directed toward HZE-induced alterations both on non-dividing systems, such as the retina, cornea and brain, and on dividing systems, such as the gut and testis. This paper is focused on the morphologically detectable late-occurring alterations in the distal lung, and toward a comparison of the changes with those induced by x-irradiation. Briefly, the salient alterations involve an increase in the width of the septal walls and the capillary and alveolar basal laminae, and the irregularity of the luminal surface of the capillaries, as exemplified by the presence of filipodial projections and blebbing. All alterations were focal in their localization, and no cells of any type (e.g., epithelial, endothelial or stromal) appeared to undergo damage, an observation quite unlike the cellular changes induced by x-irradiation.

Cardiac muscle ultrastructure and cyclic AMP reactions to altered gravity conditions.

Morphological and biochemical analyses of heart muscle of rats subjected to microgravity on Spacelab 3 (SL-3) flight and rats born and reared under increased gravity (1.7 G) conditions were compared with 1-G controls. Electronmicroscopic studies showed an increase in the number of lipid droplets and in areas of glycogen storage. Distribution changes of microtubules and cytoskeletal elements from both SL-3 and 1.7-G groups were observed. The high Km cyclic AMP phosphodiesterase activity was lower (P less than 0.05) in SL-3 heart muscle, and low Km activity was lower in 1.7-G males but was unaltered in females. Cyclic AMP-dependent protein kinase (cA-PK) activity was decreased in subcellular fractions of heart muscle of SL-3 animals. Recompartmentalization of cA-PK activity occurred in particulate tissue fraction of 1.7-G animals (70.3% of total for 1.7 G vs. 35.9% for controls). Phosphorylation of endogenous low-mobility proteins increased in SL-3 heart-soluble fractions. Photoaffinity labeling (18 h, 4 degrees C) decreased in type II cA-PK regulatory (R) subunits in both SL-3 and in 1.7-G male heart tissue particulate fractions. The 1.7-G female heart R subunit distribution did not differ from controls. These findings indicate that in heart muscle altered gravity conditions influenced physiological reactions similar to catecholamine-induced receptor-mediated hormonal responses.

Response of the seminiferous epithelium of the mouse exposed to low dose high energy (HZE) and electromagnetic radiation.

This study was undertaken to observe the response of a rapidly dividing cell population (spermatogonial) to Helium and Argon ions as compared to x-rays. Low doses (below 100 rads) were used to more nearly simulate radiation encountered during space missions. The methods used proved compatible for both light and electron microscope studies. The average relative biological effectiveness (RBE) for Helium ions is 1, while Argon is twice as effective in killing spermatogonial cells. Part of this mixed population of cells exhibits a higher sensitivity to radiation below 15 rads. Quantitation of the radiation effects by counting the necrotic cells is not feasible because of their rapid removal, therefore all measurements were done using the surviving fraction (S/So) of spermatogonial cells.