IFN-gamma does not mimic the catabolic effects of TNF-alpha

Cachexia is common in chronic inflammatory diseases and is attributed, in part, to an elevation of circulating proinflammatory cytokines. TNF-alpha is the prototype in this category. IFN-gamma is also thought to play a role, but the evidence supporting this model is primarily indirect. To determine the direct effects of IFN-gamma stimulation on muscle cells, we selected key components of the procatabolic signaling pathways by which TNF-alpha stimulates protein loss. We tested two hypotheses: 1) IFN-gamma mimics TNF-alpha signaling by increasing intracellular oxidant activity and activating MAPKs and NF-kappaB and 2) IFN-gamma increases the expression of the ubiquitin ligases atrogin1/MAFbx and muscle-specific ring finger protein 1 (MuRF1). Results showed that treatment with IFN-gamma at 60 ng/ml increased Stat1 phosphorylation after 15 min, indicating receptor activation. IFN-gamma had no effect on cytosolic oxidant activity, as measured by 2',7'-dichlorofluorescein oxidation. Nor did IFN-gamma activate JNK, ERK1/2, or p38 MAPK, as assessed by Western blot. Treatment for up to 60 min did not decrease IkappaB-alpha protein levels, as measured by Western blot analysis, or the DNA binding activity of NF-kappaB, as measured by EMSA. After 6 h, IFN-gamma decreased Akt phosphorylation and increased atrogin1/MAFbx and MuRF1 mRNA. Daily treatment for up to 72 h did not alter adult fast-type myosin heavy chain content or the total protein-to-DNA ratio. These data show that responses of myotubes to IFN-gamma and TNF-alpha differ markedly and provide little evidence for a direct catabolic effect of IFN-gamma on muscle.     

Repeated psychosocial stress at night,but not day,affects the central molecular clock

We have recently demonstrated that the outcome of repeated social defeat (SD) on behavior, physiology and immunology is more negative when applied during the dark/active phase as compared with the light/inactive phase of male C57BL/6 mice. Here, we investigated the effects of the same stress paradigm, which combines a psychosocial and novelty stressor, on the circadian clock in transgenic PERIOD2::LUCIFERASE (PER2::LUC) and wildtype (WT) mice by subjecting them to repeated SD, either in the early light phase (social defeat light?=?SDL) or in the early dark phase (social defeat dark?=?SDD) across 19 days. The PER2::LUC rhythms and clock gene mRNA expression were analyzed in the suprachiasmatic nucleus (SCN) and the adrenal gland, and PER2 protein expression in the SCN was assessed. SDD mice showed increased PER2::LUC rhythm amplitude in the SCN, reduced Per2 and Cryptochrome1 mRNA expression in the adrenal gland, and increased PER2 protein expression in the posterior part of the SCN compared with single-housed control (SHC) and SDL mice. In contrast, PER2::LUC rhythms in the SCN of SDL mice were not affected. However, SDL mice exhibited a 2-hour phase advance of the PER2::LUC rhythm in the adrenal gland compared to SHC mice. Furthermore, plasma levels of brain-derived neurotrophic factor (BDNF) and BDNF mRNA in the SCN were elevated in SDL mice. Taken together, these results show that the SCN molecular rhythmicity is affected by repeated SDD, but not SDL, while the adrenal peripheral clock is influenced mainly by SDL. The observed increase in BDNF in the SDL group may act to protect against the negative consequences of repeated psychosocial stress.     

Immobilization on the day 14th does not disrupts the basic diurnal rhythm of bone resorption

Weight bearing and physical activity are important mechanical stimuli to bone growth and metabolism, and microgravity, such a space flight and/or bed rest, induces bone resorption and bone loss. An increased excretion of urinary Ca, an increased bone resorption and a decreased bone mineral density (BMD) have been observed in bed rest experiment of healthy subjects. Bone resorption markers show the specific circadian rhythms in human. Cross-linked carboxyl-terminal telopeptide of type I collagen (ICTP) and the urinary excretion of deoxypyridinoline (Dpy) are the highest in the early morning and the lowest late at night. Bed rest immobilization might influence these rhythms, due to no mechanical loading with loss of daily life activity. Bone resorption markers in healthy subjects had been compared between before and during bed rest to determine disruption of diurnal rhythms of bone resorption.     

Rapid reset of the corticosterone rhythm by food presentation in rats under acircadian restricted feeding schedule

Corticosterone levels were determined in the 7-week-old male rat maintained under different feeding and lighting schedules. At 4 weeks of age, the animals were kept either under a natural photoperiod (LD) or were subjected to continuous illumination (LL). Access to food was either ad libitum or restricted to an 8 hr span per 24 hr (circadian) or 32 hr (acircadian).

The food signal seemed able to synchronize the corticosterone rhythm to its own circadian periodicity, irrespective of the lighting regimen. No synchronization was observed in serially sampled LL or LD rats under an acircadian feeding schedule. Instead, the group acrophase appeared 24 hr subsequent to food presentation. Regarding individual patterns, many rats showed an acrophase or a peak also at that time. We speculate that an endogenous circadian mechanism was reset by the food signal, whenever it appeared.     

The role of the daily feeding rhythm in the regulation of the day/night rhythm in triglyceride secretion in rats

Plasma triglyceride (TG) levels show a clear daily rhythm, however, thus far it is still unknown whether this rhythm results from a daily rhythm in TG production, TG uptake or both. Previous studies have shown that feeding activity affects plasma TG concentrations, but it is not clear how the daily rhythm in feeding activity affects plasma TG concentrations. In the present study, we measured plasma TG concentrations and TG secretion rates in rats at 6 Zeitgeber times to investigate whether plasma TG concentrations and TG secretion show a daily rhythm. We found that plasma TG concentrations and TG secretion show a significant day/night rhythm. Next, we removed the daily rhythm in feeding behavior by introducing a 6-meals-a-day (6M) feeding schedule to investigate whether the daily rhythm in feeding behavior is necessary to maintain the daily rhythm in TG secretion. We found that the day/night rhythm in TG secretion was abolished under 6M feeding conditions. Hepatic apolipoprotein B (ApoB) and microsomal TG transfer protein (Mttp), which are both involved in TG secretion, also lost their daily rhythmicity under 6M feeding conditions. Together, these results indicate that: (1) the daily rhythm in TG secretion contributes to the formation of a day/night rhythm in plasma TG levels and (2) a daily feeding rhythm is essential for maintaining the daily rhythm in TG secretion.     

The effects of stress on plasma ACTH and corticosterone in young and aging pregnant rats and their fetuses

Compared to younger rats, old rats exhibit prolonged elevations of plasma ACTH and corticosterone (CORT) in response to stress. In addition, CORT crosses the placenta. To investigate whether fetuses of older rats may be exposed to higher concentrations of CORT during development than fetuses of young rats, we compared the effects of stress on hypothalamic-pituitary-adrenal (HPA) axis function in young and aging pregnant rats and their 19-day-old fetuses. The plasma of the mothers and fetuses was assayed for ACTH and CORT by radioimmunoassay. Both young and aging pregnant rats showed a significant increase in plasma ACTH and CORT immediately after exposure to stress. However, aging rats had more prolonged elevations of ACTH and CORT than young rats. This suggests that, like old male rats, aging pregnant rats have an alteration in feedback inhibition of the HPA axis. Prolonged elevation of CORT was also seen in fetuses of aging mothers. These results have important implications concerning the effects of stress during pregnancy at different maternal ages, and for the potential deleterious consequences of prolonged prenatal elevation in stress hormones on the offspring of aging females.     

Dietary tyrosine suppresses the rise in plasma corticosterone following acute stress in rats

Acute, uncontrollable stress increases norepinephrine (NE) turnover in the rat's brain (depleting NE) and diminishes the animal's subsequent tendency to explore a novel environment. Pre-treatment with tyrosine can reverse these adverse effects of stress, presumably by preventing the depletion of NE in the hypothalamus. Numerous studies suggest that NE inhibits the release of adrenocorticotropic hormone (ACTH) by suppressing corticotropic releasing factor (CRF) secretion in the hypothalamus. In the present study, we found that pre-treatment with supplemental tyrosine not only prevented the behavioral depression and hypothalamic NE depletion observed after an acute stress, but also suppressed the rise in plasma corticosterone. These results support a role for brain NE in stress-induced corticosterone secretion and demonstrate that supplemental tyrosine can protect against several adverse consequences of such stress.     

Influence of the corticosterone rhythm on photic entrainment of locomotor activity in rats

The question of involvement of glucocorticoid hormones as temporal signals for the synchronization of the timekeeping system was addressed in rats with different corticosterone status. The authors showed that adrenalectomy had no effects on the synchronization of wheel-running activity rhythms to a steady-state LD 12:12 cycle, regardless of whether it was compensated for by a corticosterone replacement therapy that either reinstated constant plasma concentrations of the hormone or mimicked its natural rhythm. However, after a 12-h phase shift (daylight reversal), the lack of circulating corticosterone induced a significant shortening of the resynchronization rate (less than 3 days vs. 7 days). Normalization required restoration of a rhythmic corticosterone secretion that was synchronized to the new photoperiod. Under constant darkness, the corticosterone rhythm did not show any synchronizing effect, providing evidence that it participates in entrainment of the locomotor activity rhythm through modulation of light effects. It is proposed that, under stable lighting conditions, circulating glucocorticoids contribute to stabilizing activity rhythms by reinforcing resistance of the circadian timing system to variations of the photoperiod. Experimental evidence that serotonergic neurons are involved in relaying their modulatory effects to the clock is also presented.     

ACTH, corticosterone and beta-endorphin in the blood plasma of rats under prolonged immobilization stress

Radioimmunoassays were employed to study variation in the concentration of corticosterone, ACTH and beta-endorphin in rat blood plasma at different times of the 30-hour immobilization and under the effect of a short-term action of ecologically significant negative stimulation emanating from other animals. Both prolonged immobilization stress and short-term emotional reaction produced appreciable alterations in the blood plasma content of all the hormones under study. The findings indicate that variations in the corticosterone and beta-endorphin levels were in the best agreement.     

Circaseptan rhythm of corticosterone and lipids in the serum of male rats

After adaptation to standard conditions and to a 12:12 h light: dark regimen, groups of young SPF male Wistar rats were killed at 8 h on 17 consecutive days and their serum corticosterone and lipid concentrations were determined. Using a combination of a Fourier harmonic analysis and an analysis of variance, the circaseptan rhythm of corticosterone, total cholesterol and triacylglycerol oscillations was determined; it is unlikely that the nonesterified fatty acid concentration has a 7-day rhythm. The circaseptan variation of metabolic and hormonal indicators is evidently adaptive in character and its existence ought to be taken particularly into account in the evaluation of long-term experiments.     

Abiotic stress does not magnify the deleterious effects of spontaneous mutations

Although the effects of deleterious alleles often are predicted to be greater in stressful environments, there is no theoretical basis for this prediction and the empirical evidence is mixed. Here we characterized the effects of three types of abiotic stress (thermal, oxidative and hyperosmotic) on two sets of nematode (Caenorhabditis elegans) mutation accumulation (MA) lines that differ by threefold in fitness. We compared the survival and egg-to-adult viability between environments (benign and stressful) and between fitness categories (high-fitness MA, low-fitness MA). If the environment and mutation load have synergistic effects on trait means, then the difference between the high and low-fitness MA lines should be larger in stressful environments. Although the stress treatments consistently decreased survival and/or viability, we did not detect significant interactions between fitness categories and environment types. In contrast, we did find consistent evidence for synergistic effects on (micro)environmental variation. The lack of signal in trait means likely reflects the very low starting fitness of some low-fitness MA lines, the potential for cross-stress responses and the context dependence of mutational effects. In addition, the large increases in the environmental variance in the stressful environments may have masked small changes in trait means. These results do not provide evidence for synergism between mutation and stress.     

Daily rhythm of plasma corticosterone binding activity in the white-throated sparrow,Zonotrichiaalbicollis

There is a bimodal daily rhythm of plasma corticosterone binding activity (CBA) in the white-throated sparrow, $\text{Zonotrichia}$$\text{albicollis}$, in the spring migratory condition. Low levels occurred at the beginning (0500) and at the end (2100) of a 16-hour daily photoperiod. Peak CBA occurred at 0900 and 0100, as did peak locomotor activity in this nocturnal migrant. Comparisons of CBA with total plasma corticosteroid concentrations from a previous study of the same group of birds indicate a positive correlation during most of the day but not during the early hours of darkness. The daily rhythm of locomotor activity may account for the rhythm of CBA which, in turn, may be partially responsible for the daily rhythm of plasma corticosteroid concentration.     

Adrenal splanchnic innervation contributes to the diurnal rhythm of plasma corticosterone in rats by modulating adrenal sensitivity to ACTH

Activity of the hypothalamic-pituitary-adrenal axis is characterized by a diurnal rhythm with an AM nadir and PM peak. Splanchnic nerve transection disrupts the diurnal rhythm in plasma corticosterone; however, there is a controversy as to whether the nerve-mediated effect is 1) via inhibition in the AM vs. excitation in the PM, or 2) involves changes in adrenal sensitivity to ACTH. The present studies were designed to address these issues. Adult male rats were anesthetized and underwent bilateral transection of the thoracic splanchnic nerve or sham transection. One week after surgery, rats were killed in the AM or PM with collection of nonstress plasma for measurement of corticosterone and ACTH. Plasma corticosterone was increased in the PM relative to the AM; however, plasma corticosterone in the PM was attenuated by splanchnic nerve transection, without affecting plasma ACTH. This decrease in PM plasma corticosterone after nerve-transection was 1) associated with decreased adrenal responsivity to ACTH, 2) associated with decreased adrenal cAMP content, 3) prevented by adrenal demedullation, and 4) not affected by removal of adrenal capsaicin-sensitive afferent fibers. Repeated serial blood sampling from individual rats confirmed the excitatory effect of splanchnic innervation in the PM. These results support the hypothesis that the adrenal splanchnic innervation modulates the diurnal rhythm in plasma corticosterone by increasing adrenal responsivity to ACTH and augmenting steroidogenesis in the PM and suggest that alterations in adrenal corticosterone secretion obscured by pulsatile secretion are more clearly revealed with repeated serial blood sampling.     

Anion binding at the active site of trypanosomal triosephosphate isomerase. Monohydrogen phosphate does not mimic sulphate

The three-dimensional structure of triosephosphate isomerase complexed with the competitive inhibitor SO-4(2) was determined by X-ray crystallography to a resolution of 0.24 nm. A comparison with the native crystal structure, where SO-4(2) is bound, revealed five changes: (a) a 0.10-nm shift of the anion-binding site; (b) a further closing of the flexible loop of the enzyme; (c) a 'swinging in' of the side chain of the catalytic Glu, that is chi 1 changes from (+) to (-) synclinal; (d) an altered water structure; (e) a disappearance of the conformational heterogeneity at the C-terminus of strand beta 7. Some of these changes may be related to the different hydrogen-bond pattern about the two different anions. However, the distance of 0.10 nm between the sulphur and phosphorus positions is unexpected and remains intriguing.     

The effect of strength training at the same time of the day on the diurnal fluctuations of muscular anaerobic performances

The aim of this study was to examine the effects of training at the same time of the day on the diurnal variations of anaerobic performances to provide some recommendations to adjust training hours with the time of the day of competitive events. Thirty participants underwent 8 weeks of lower-extremity progressive resistance training performed 3 times per week designed to promote muscular strength and power. These subjects were randomly assigned to a morning training group (MTG, 07:00-08:00 hours, n = 10), an evening training group (ETG, 17:00-18:00 hours, n = 10), and a control group (CG, completed all tests but did not train, n = 10). Performance in the squat jump, the countermovement jump, the Wingate and 1 repetition maximum (1RM) during leg extension, leg curl, and squat tests was recorded just before and 2 weeks after an 8-week course of regular training. For all the subjects, the morning and evening tests were scheduled at the same time of the day as for the morning and evening training sessions. Before training, the results indicated a significant increase in performance from morning to evening tests (ca. 2.84-17.55% for all tests) for all groups. After training, the diurnal variations in anaerobic performances were blunted in the MTG. In fact, there was no significant difference in muscular power or strength between morning and evening tests. However, these intradaily variations in anaerobic performances persisted in the ETG and CG. From a practical point of view, adaptation to strength training is greater at the time of the day at which training was scheduled than at other times.     

Factors modifying the effect of diazepam on plasma corticosterone levels in rats

We have examined factors that alter the effect of diazepam (DZ) on plasma corticosterone (CS) in rats. DZ had a biphasic effect on plasma CS levels: CS decreased with doses below 5 mg/kg and increased with higher doses. Peak response occurred 90 minutes post injection in both sexes. Plasma DZ levels were significantly higher in females than in males and peak at 10 and 30 minutes post injection in males and females, respectively. There was also a sex difference in the pattern of DZ metabolites. An acute stressor (30 minutes of immobilization) did not affect plasma CS levels in rats injected with a 5 mg/kg dose of DZ. Prenatally stressed animals did not differ in basal CS levels or in their response to 5 mg/kg of DZ compared to prenatally non-stressed animals. These two groups of animals also did not differ in plasma levels of DZ or of its metabolites. By contrast, the 5 mg/kg dose of DZ had no effect on plasma testosterone levels in control animals, but increased it in prenatally stressed animals. Furthermore, compared to non-stressed controls, prenatally stressed animals had lower baseline plasma testosterone levels. These results indicate that the effect of DZ on plasma CS is influenced by endogenous as well as exogenous factors and that these effects vary with the particular biochemical parameter under examination.