The effect of 2G on mouse circadian rhythms.

This study examined the effect of chronic 2G exposure on the regulation of body temperature (T(b)), activity (ACT), and circadian rhythms of mice. Five mice were implanted with biotelemetry units to record T(b) and ACT. The mice exhibited a stable daily mean of T(b) (37.1 +/- 2.1 degrees C) and ACT and robust circadian rhythms during the control 1G period. Mice exhibited a significant decline in T(b) (30.1 +/- 1.5 degrees C; t(4)=8.32, p<.01) and cessation of ACT within two hours following 2G onset. After 6 hours of continuous 2G exposure there was a recovery in T(b) (34.4 +/- 1.6 degrees C) that remained significantly below that of baseline (t(4)=3.66, p<.05). A similar pattern of recovery was seen following 12 hours of continuous 2G for ACT. A slower pattern of adaptation toward baseline levels occurred steadily over the next 6-13 days. Exposure to 2G also caused an immediate 4 day loss in circadian rhythm amplitude in both T(b) and ACT. Recovery to new steady state levels was achieved by 8 days and 13 days, respectively. These results demonstrate that under chronic 2G, the recovery time for the homeostatic steady-state values and circadian rhythms are shorter for the mouse than for the rat. These differences may be related to the scaling effects of 2G resulting from the mass difference between mice and rats.

Effects of square-wave and simulated natural light-dark cycles on hamster circadian rhythms.

Circadian rhythms of activity (Act) and body temperature (Tb) were recorded from male Syrian hamsters under square-wave (LDSq) and simulated natural (LDSN, with dawn and dusk transitions) light-dark cycles. Light intensity and data sampling were under the synchronized control of a laboratory computer. Changes in reactive and predictive onsets and offsets for the circadian rhythms of Act and Tb were examined in both lighting conditions. The reactive Act onset occurred 1.1 h earlier (P < 0.01) in LDSN than in LDSq and had a longer alpha-period (1.7 h; P < 0.05). The reactive Tb onset was 0.7 h earlier (P < 0.01) in LDSN. In LDSN, the predictive Act onset advanced by 0.3 h (P < 0.05), whereas the Tb predictive onset remained the same as in LDSq. The phase angle difference between Act and Tb predictive onsets decreased by 0.9 h (P < 0.05) in LDSN, but the offsets of both measures remained unchanged. In this study, animals exhibited different circadian entrainment characteristics under LDSq and LDSN, suggesting that gradual and abrupt transitions between light and dark may provide different temporal cues.

Development of circadian rhythms in rat pups exposed to microgravity during gestation.

Ten pregnant Sprague Dawley rat dams were exposed to spaceflight aboard the Space Shuttle (STS-70) for gestational days 11-20 (G 11-20; FLT group). Control dams were maintained in either a flight-like (FDS group) or vivarium cage environment (VIV group) on earth. All dams had ad lib access to food and water and were exposed to a light-dark cycle consisting of 12 hours of light (approximately 30 lux) followed by 12 hours of darkness. The dams were closely monitored from G 22 until parturition. All pups were cross-fostered at birth; each foster dam had a litter of 10 pups. Pups remained with their foster dam until post natal day 21 (PN 21). Pup body mass was measured twice weekly. At PN 14 FLT pups had a smaller body mass than did the VIV pups (p < 0.01). Circadian rhythms of body temperature and activity of pups from two FLT dams (n = 8), two FDS dams (n = 9) and two VIV dams (n = 7) were studied starting from age PN 21. All pups had circadian rhythms of temperature and activity at this age. There were no significant differences in rhythms between groups that could be attributed to microgravity exposure. These results indicate that exposure to the microgravity environment of spaceflight during this embryonic development period does not affect the development of the circadian rhythms of body temperature and activity.

Chronic 2G exposure affects c-Fos reactivity to a light pulse within the rat suprachiasmatic nucleus.

This study examined the effect of the hyperdynamic environment on the function of the retinohypothalamic tract. Rats were exposed to either 2 days or 21 days of 2G via centrifugation. During the last hour of 2G exposure, one series of rats was exposed to a 1 hour phase-shifting light pulse while the second series of rats did not receive a light pulse. In addition a groups of 1G controls was exposed to the same 1 hour lighting paradigm. All animals were processed for c-Fos within the SCN. The 1G controls showed the normal response to light in which significantly greater numbers of c-Fos positive neurons were found in the SCN of the light pulsed rats relative to that of the nonlight pulsed rats. However, rats exposed to 2 days of 2G did not show the same response to light. Light pulsed rats and nonlight pulsed rats exhibited few c-Fos positive neurons within the SCN. A recovery in the effect of light to induce c-Fos reactivity within SCN neurons occurred in the rats exposed to 21 days of 2G. These results suggest that exposure to 2G can temporarily suppress the responsiveness of the SCN to the phase-shifting effects of light mediated by the retinohypothalamic tract.

The effect of a BRN 3.1 deletion on the temperature response to 2G.

NASA: Researchers studied the effect of 2G exposure on body temperature in Wild type and BRN 3.1 Knockout mice to determine the feasibility to using BRN 3.1 Knockout mice as an animal model of the effects of altered gravitational fields on vestibular system physiology.^ieng

The effect of spaceflight on retino-hypothalamic tract development.

NASA: Researchers examined the effect of late prenatal exposure to microgravity on the development of the retina, retinohypothalamic tract, geniculo-hypothalamic tract, and suprachiasmatic nucleus. Results indicate an effect on c-fos activity in the intergeniculate leaflet between gestational day 20 and postnatal day 8, suggesting a delay in development of the circadian timing system.^ieng

Circadian rhythms of temperature and activity in obese and lean Zucker rats.

The circadian timing system is important in the regulation of feeding and metabolism, both of which are aberrant in the obese Zucker rat. This study tested the hypothesis that these abnormalities involve a deficit in circadian regulation by examining the circadian rhythms of body temperature and activity in lean and obese Zucker rats exposed to normal light-dark cycles, constant light, and constant dark. Significant deficits in both daily mean and circadian amplitude of temperature and activity were found in obese Zucker female rats relative to lean controls in all lighting conditions. However, the circadian period of obese Zucker rats did not exhibit differences relative to lean controls in either of the constant lighting conditions. These results indicate that although the circadian regulation of temperature and activity in obese Zucker female rats is in fact depressed, obese rats do exhibit normal entrainment and pacemaker functions in the circadian timing system. The results suggest a deficit in the process that generates the amplitude of the circadian rhythm.

Acute exposure to 2G phase shifts the rat circadian timing system.

The circadian timing system (CTS) provides internal and external temporal coordination of an animal's physiology and behavior. In mammals, the generation and coordination of these circadian rhythms is controlled by a neural pacemaker, the suprachiasmatic nucleus (SCN), located within the hypothalamus. The pacemaker is synchronized to the 24 hour day by time cues (zeitgebers) such as the light/dark cycle. When an animal is exposed to an environment without time cues, the circadian rhythms maintain internal temporal coordination but exhibit a "free-running" condition in which the period length is determined by the internal pacemaker. Maintenance of internal and external temporal coordination are critical for normal physiological and psychological function in human and non-human primates. Exposure to altered gravitational environments has been shown to affect the amplitude, mean, and timing of circadian rhythms in species ranging from unicellular organisms to man. However, it has not been determined whether altered gravitational fields have a direct effect on the neural pacemaker, or affect peripheral physiological systems that express these circadian parameters. In previous studies, the ability of a stimulus to phase shift circadian rhythms was used to determine whether a stimulus has a direct effect on the neural pacemaker. The present experiment was performed in order to determine whether acute exposure to a hyperdynamic field could phase shift circadian rhythms.

Changes in hypothalamic [correction of hypothalmic] staining for c-Fos following 2G exposure in rats.

The static gravitational field of the earth has been an important selective pressure that has shaped the evolution of biological organisms. This is illustrated by the evolution of tetrapods from a water environment where gravitational force was partially negated to a terrestrial environment where gravity is of greater consequence. Terrestrial invasion resulted in a series of new structural, physiological, and behavioral features. Therefore, it is not surprising that alterations in the gravitational field can cause widespread effects in many physiological systems and behaviors. Our previous studies have demonstrated that both exposure to hyperdynamic fields and the microgravity condition of space flight have significant effects on body temperature, heartrate, activity, feeding, drinking, and circadian rhythms. However, it has not been determined whether these physiological adaptations are associated with changes in neural activity within the hypothalamic nuclei that regulate these functions. This study examined the changes in body temperature, activity, body weight and food and water intake in rats caused by exposure to a hyperdynamic field. In addition, the immediate early gene activation marker, c-Fos, was used to examine potential protein synthesis changes in the hypothalamic nuclei that regulate these functions.

Influence of gravity on the circadian timing system.

The circadian timing system (CTS) is responsible for daily temporal coordination of physiological and behavioral functions both internally and with the external environment. Experiments in altered gravitational environments have revealed changes in circadian rhythms of species ranging from fungi to primates. The altered gravitational environments examined included both the microgravity environment of spaceflight and hyperdynamic environments produced by centrifugation. Acute exposure to altered gravitational environments changed homeostatic parameters such as body temperature. These changes were time of day dependent. Exposure to gravitational alterations of relatively short duration produced changes in both the homeostatic level and the amplitude of circadian rhythms. Chronic exposure to a non-earth level of gravity resulted in changes in the period of the expressed rhythms as well as in the phase relationships between the rhythms and between the rhythms and the external environment. In addition, alterations in gravity appeared to act as a time cue for the CTS. Altered gravity also affected the sensitivity of the pacemaker to other aspects of the environment (i.e., light) and to shifts of time cues. Taken together, these studies lead to the conclusion that the CTS is indeed sensitive to gravity and its alterations. This finding has implications for both basic biology and space medicine.

Bone contusions of the knee: increased lesion detection with fast spin-echo MR imaging with spectroscopic fat saturation.

PURPOSE: To demonstrate that T2-weighted fast spin-echo (FSE) imaging with spectroscopic fat saturation (FS-FSE) increases the conspicuity between normal marrow and bone contusions in posttraumatic knees. MATERIALS AND METHODS: Seventy-six magnetic resonance (MR) studies of the knee were prospectively evaluated in 73 consecutive patients with knee pain. Conspicuity of regions of microtrabecular trauma (bone contusions) was evaluated with conventional T2-weighted imaging in the sagittal plane, T2-weighted FSE imaging in the coronal plane, T2-weighted FS-FSE imaging in the sagittal plane, and conventional T1-weighted imaging in the sagittal plane. RESULTS: Twenty-six foci of bone contusion were identified in 21 knees with the FS-FSE technique. Only 16 foci were demonstrated with conventional T2-weighted imaging. Six of the sites of bone contusion were not demonstrated with the FSE technique alone (without fat saturation). Four of the bone contusions were not seen on T1-weighted images; these sites of bone contusion were substantially more conspicuous on the FS-FSE images. In addition, FS-FSE imaging was more sensitive in demonstrating the extent of microtrabecular trauma. CONCLUSION: T2-weighted FS-FSE imaging is a sensitive and rapid method of identifying and assessing the extent of microtrabecular trauma about the knee.

The regulation of rat activity following exposure to hyperdynamic fields.

The microgravity of spaceflight and the hyperdynamic fields produced via centrifugation have allowed researchers to examine the effect of altered gravitational environments on the regulation of physiological systems. In previous studies, we have discussed the importance of homeostatic and circadian mechanisms for the regulation of physiological systems such as body temperature and heart rate. Rats exposed to a chronic 2G field exhibited lower mean daily body temperature and heart rate. The homeostatic component of regulation for body temperature and heart rate adapts to a new steady state after 5-6 days. However, the circadian rhythm of body temperature and heart rate became severely depressed and did not recover for approximately 7-10 days. The measurements of body temperature and heart rate suggest an adaptation of homeostatic and circadian regulatory mechanisms following 10 days exposure to 2G. However, an important function of physiological homeostasis is to respond to environmental stressors. An important question thus becomes whether the regulation of body temperature and heart rate has sufficiently recovered to respond to an environmental challenge separate from that of the hyperdynamic field. In this study, a high frequency light/dark cycle (LD 3:3) was provided for 24 hours as an environmental challenge to assess the recovery of homeostatic and circadian regulation. Previous studies have demonstrated that high frequency light dark cycles are highly effective for testing homeostatic and circadian components of physiological regulation in monkeys and rats. For example, the nocturnal rat exhibited a homeostatic increase in body temperature during the dark periods and a decrease during the light periods. In addition, the magnitude of the body temperature response exhibits a time of day variation demonstrating the effect on circadian regulation.

Advances in imaging of rheumatoid arthritis.

Several new imaging modalities have been found useful in clinical evaluation of patients with rheumatoid arthritis (RA). Magnetic resonance (MR) imaging has proven to be an excellent noninvasive method to evaluate the spine, shoulder, hip, and knees; its use for the evaluation of smaller joints is still being investigated. In patients with RA, MR imaging has been used to evaluate cervical spine instability, rotator cuff tear, osteonecrosis, and osteomyelitis. Patients with RA may have advanced osteoporosis, predisposing to insufficiency fractures. This includes fractures associated with increased activity after hip or knee arthroplasty. Newer methods for measuring the degree of osteoporosis include single photon absorptiometry, dual photon absorptiometry, quantitative computed tomography (CT), and dual-energy projection radiography. It has not yet been determined which of these methods will be most widely used in the future, but quantitative CT and dual-energy projection radiography currently show the most promise. Ultrasonography provides an excellent noninvasive method for the diagnosis of popliteal cysts, and color Doppler sonography can differentiate cyst and popliteal aneurysm. As compared to radiography or conventional CT, high-resolution CT provides an improved method to detect the early changes of RA in the lung parenchyma.

Neuronal activity in hypothalamic nuclei of obese and lean Zucker rats.

Central neural activity was assessed by measuring relative cytochrome oxidase (CO) activity in the ventromedial nucleus (VMN; thermogenesis regulation), the parvocellular paraventricular nucleus (PVN; feeding regulation), and the magnocellular PVN (secretion of vasopressin and oxytocin) in 10 age-matched pairs of 39- to 42-day-old Zucker rats. When obese (fa/fa) were compared to lean (Fa/Fa) rats, relative CO activity was significantly lower (approximately 10 percent) in the VMN and parvocellular PVN, but not in the magnocellular PVN. Cell diameters did not differ. To determine if there were corresponding differences in levels or release of hypothalamic monoamines, we compared 7 pairs of 90- to 94-day-old lean (Fa/?) and obese (fa/fa) rats at rest and after 2 h of 9 degrees C. Tissue punches from frozen PVN, VMN, and preoptic area (the latter being a site of thermosensitive units modulating VMN output) were assayed. In obese vs. lean noncold-exposed rats, we observed lower concentrations of: 5-hydroxyindoleacetic acid (5HIAA; metabolite of serotonin, 5HT) in the VMN; 3-methoxy-4-hydroxyphenylglycol (MHPG; metabolite of norepinephrine, NE) and NE + MHPG (index of total NE) in the preoptic area; and 3,4-dihydroxyphenylacetic acid (DOPAC; metabolite of dopamine, DA) in the PVN. Additionally, in the VMN, cold exposure resulted in: elevated concentrations of MHPG and MHPG + NE in both lean and obese rats; elevated concentrations of 5HT, 5HIAA, and 5HT + 5HIAA in obese rats, with no significant changes in these variables in lean animals; decreased ratio of 5HIAA/5HT in obese rats and increased ratio in leans. In the preoptic region, cold exposure led to increased concentrations of MHPG, NE + MHPG, 5HT, and 5HT + 5HIAA in obese but not lean rats. In the PVN, 5HT concentrations were increased in cold-exposed obese but not lean rats. Our data support the hypothesis that neuronal activity in obese rats differs from that of lean rats at rest and during cold exposure and suggest that several monoamine systems play a role in such differences.

The effect of hyperdynamic fields on the oxidative metabolism of the paraventricular nucleus.

An important issue in space biology and medicine is understanding the effect of gravitational changes on the mechanisms that regulate fluid homeostasis. The results of this study show that following 7 d exposure to a 2 G or 3 G hyperdynamic field, rats exhibited a linear increase in the cytochrome oxidase staining of neurons in the paraventricular nucleus (PVN). The elevated oxidative metabolism in the PVN suggests that there was an increase in the manufacturing and release of vasopressin into the plasma in response to a perceived hypovolemic condition caused by increased hydrostatic pressure and redistribution of fluid to the periphery. Since vasopressin also has widespread cardiovascular effects, it will be important to understand the relationship between vasopressin and altered gravitational fields.

The retinohypothalamic projection and oxidative metabolism in the suprachiasmatic nucleus of primates and tree shrews.

This study compared the patterns of retinal projections and oxidative metabolism in the hypothalamus of squirrel monkeys, Bonnet macaques, and tree shrews. Intraocular injections of horseradish peroxidase in primates demonstrated that retinal terminals were present from the anterior to posterior poles of the suprachiasmatic nucleus (SCN). The terminals were primarily located in the ventral and ventrolateral regions of the SCN. In addition, there was a relatively even density and distribution of retinal terminals between the ipsilateral and contralateral projections. The pattern of oxidative metabolism in the hypothalamus of the primates examined demonstrated that the SCN is highly metabolic relative to the surrounding area, and distinct regions of the SCN exhibit clear differences in metabolism. These distinct metabolic regions may reflect functional subdivisions within the SCN. In addition, elevated metabolism is found along the hypothalamo-optic chiasm border. The retinal projection to the hypothalamus in tree shrews was very different from that of the primates examined. The contralateral retinal projection was very dense, but the ipsilateral retinal projection was very sparse. Retinal terminals were primarily distributed along the lateral border of the SCN. Both the SCN and the region lateral to the SCN exhibited elevated oxidative metabolism relative to the surrounding hypothalamus.

The retinohypothalamic tract in the cat: retinal ganglion cell morphology and pattern of projection.

The pattern of retinal projection to the hypothalamus and the morphological properties of the retinal ganglion cells that comprise the retinohypothalamic tract have been examined in the cat. Intraocular injections of horseradish peroxidase revealed a dense retinal projection to the ventral suprachiasmatic nucleus; however, lighter projections were seen in the dorsal suprachiasmatic nucleus, and in hypothalamic regions both dorsal and lateral to the suprachiasmatic nucleus. Intrasuprachiasmatic nucleus injections of horseradish peroxidase retrogradely labelled retinal ganglion cells that were small to medium in soma size. The labelled ganglion cells exhibited long thin dendrites that were sparsely branched. The labelled retinal ganglion cells exhibited a significant change in soma size associated with retinal eccentricity. The morphological characteristics of the ganglion cells that project to the suprachiasmatic nucleus are similar to those of gamma cells.

Temperature regulation in rats exposed to a 2 G field.

The regulation of body temperature involves both homeostatic and circadian control systems. Both systems are influenced by exposure to hyperdynamic fields and demonstrate acute responses that eventually recover to an adapted level. This experiment examined both the homeostatic and circadian responses of body temperature to a separate environmental challenge (high frequency light/dark cycles) during exposure to a 2 G hyperdynamic field.