Pain inhibition and postexertional malaise in myalgic encephalomyelitis/chronic fatigue syndrome: an experimental study.

OBJECTIVES: To examine the efficacy of the pain inhibitory systems in patients with myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) during two different types of exercise and to examine whether the (mal)functioning of pain inhibitory systems is associated with symptom increases following exercise. DESIGN: A controlled experimental study. SETTING AND SUBJECTS: Twenty-two women with ME/CFS and 22 healthy sedentary controls were studied at the Department of Human Physiology, Vrije Universiteit Brussel. INTERVENTIONS: All subjects performed a submaximal exercise test and a self-paced, physiologically limited exercise test on a cycle ergometer. The exercise tests were undertaken with continuous cardiorespiratory monitoring. Before and after the exercise bouts, subjects filled out questionnaires to assess health status, and underwent pressure pain threshold measurements. Throughout the study, subjects' activity levels were assessed using accelerometry. RESULTS: In patients with ME/CFS, pain thresholds decreased following both types of exercise, whereas they increased in healthy subjects. This was accompanied by a worsening of the ME/CFS symptom complex post-exercise. Decreased pressure thresholds during submaximal exercise were associated with postexertional fatigue in the ME/CFS group (r = 0.454; P = 0.034). CONCLUSIONS: These observations indicate the presence of abnormal central pain processing during exercise in patients with ME/CFS and demonstrate that both submaximal exercise and self-paced, physiologically limited exercise trigger postexertional malaise in these patients. Further study is required to identify specific modes and intensity of exercise that can be performed in people with ME/CFS without exacerbating symptoms.

Disappearance rate of glycosylated and non-glycosylated chicken growth hormone: influence on biological activity.

The disappearance rates for glycosylated (GcGH) and non-glycosylated chicken growth hormone (NGcGH) were compared following their intravenous injection into anaesthetized adult laying hens. The metabolic clearance rate of GcGH was about 20% lower (P = 0.13) compared to NGcGH. Similarly, there was a tendency towards a longer half-life for GcGH. No major physiological significance could be attributed to the glycosylation state of the preparations as far as their effects on circulating insulin-like growth factor I and iodohormone levels were concerned.

Growth hormone secretory characteristics of sex-linked dwarf and normal-sized chickens reared on a control or on a 3,3',5-triiodothyronine-supplemented diet.

This study examined the pulsatile presence of growth hormone (GH) in the plasma--which reflects its pulsatile release--of 8-week-old male sex-linked dwarf (dwdw) or normal-sized (Dwdw) chickens from a brown-egg layer strain reared on a control or on a 0.5 ppm triiodothyronine(T3)-supplemented diet. The overall GH mean, amplitude, and baseline levels of control dw chickens were significantly higher than those of control Dw chickens. No differences in peak length or peak frequency between genotypes were observed. Dietary T3 supplementation abolished the pulsatile GH release in both genotypes. T3 treatment depressed the mean GH levels of Dw but not of dw chickens.

Plasma profiles of somatotropin and IGF-I in diary cows following application of two preparations of recombinant bovine somatotropin in a sustained release vehicle.

Milk production, plasma bovine somatotropin (bST) and insulin-like growth factor I (IGF-I) were measured in dairy cows following a single subcutaneous injection of a slowly released preparation of either recombinant enterokinase linker bST (somidobove: 640 mg) or recombinant methionyl bST (sometribove: 500 mg). There was a 3-7-fold increase in plasma bST concentrations during the first three postinjection hours in cows treated with both sometribove (from 3.4 +/- 0.8 to 11.2 +/- 3.0 ng ml-1) or somidobove (from 2.3 +/- 0.3 to 17.5 +/- 2.6 ng ml-1). In the next 8 days the bST concentration in the bST-treated cows varied, but was still significantly increased above the controls. In the following days, the concentrations of bST did not differ from the controls. Plasma concentrations of IGF-I increased nearly 2-fold as early as 24 h following recombinant bST administration and then continued to rise so that by 48 h postinjection they were nearly four times higher (control 16.2, bST-treated 61.7 ng ml-1). From 48 h after sometribove injection, IGF-I concentrations remained at a plateau (varying between 60.4 and 85.7 ng ml-1) till day 11. Then it decreased slowly, but still remained higher on day 14 than those in placebo-treated cows (44.4 +/- 17.8 ng ml-1 in bST-treated animals; 12.2 +/- 7.5 ng ml-1 in the controls). Although IGF-I level was increasing in all bST-treated animals, the absolute IGF-I increase was not related to the increase in milk production.

Plasma levels of growth hormone and insulin-like growth factor-I and -II from 2 to 6 weeks of age in meat-type chickens selected for 6-week body weight or for feed conversion and reared under high or normal environmental temperature conditions.

The aim of this study was to compare the effect of high (33 degrees C) and normal (33-20 degrees C) rearing temperature on growth and plasma levels of the somatotrophic hormones of 2 genetic lines of broiler chickens selected for 6-wk body weight (GL-line) or for feed conversion between 3 and 6 wk of age (FC-line) or for feed conversion between 3 and 6 wk of age (FC-line). Blood samples were collected weekly and analysed for growth hormone (GH) and insulin-like growth factors (IGF)-I and -II levels by RIA. The growth-depressing effect of the HT-treatment was more pronounced in the heavier GL-line and in males. A similar age-related pattern for all hormones studied was observed with the highest levels between 2 and 4 wk of age. FC-line chickens and males had consistently higher plasma GH levels than GL-line chickens and females respectively. No consistent effect of rearing temperature on plasma GH levels were observed. At 2 wk of age, HT-treatment resulted in higher plasma IGF-I levels while this was reversed from 3 wk of age onwards. GL-line chickens had significantly higher plasma IGF-I levels at 2, 3 and 4 wk of age. No consistent effect of sex on plasma IGF-I levels could be observed. For the whole period studied, GL-line chickens had significantly higher plasma IGF-II levels than FC-line chickens. No consistent effect of sex or temperature treatment on plasma IGF-II levels was observed.

Endogenous growth hormone controls high plasma levels of 3,3',5-triiodothyronine (T3) in growing chickens by decreasing the T3-degrading type III deiodinase activity.

The influence of endogenous GH levels on peripheral monodeiodination activity has been investigated in growing chickens at the age of 4 weeks, when they normally show no T3 increase after GH injection. Injection of anti-GH serum decreased plasma T3 and increased plasma T4. Three d and 1 week after hypophysectomy, plasma T3 was also markedly decreased, while T4 was only slightly affected, hepatic 5'D-I activity showed a transient decrease, but 5D-III activity was highly increased, as were the number of hepatic GH receptor sites. Injection of GH in hypophysectomized chickens decreased 5D-III activity and increased plasma T3. GH receptor-deficient dwarf chickens had decreased plasma T3 and increased plasma T4 and hepatic 5'D-I and 5D-III activities compared to their normally-growing siblings. GH administration could only affect T3 and 5D-III in the non-dwarf siblings, which showed higher basal 5D-III activity compared to the non-responsive age-matched chickens of the Hisex strain used in the other experiments. It can be concluded that endogenous GH is an important factor in the control of plasma T3 levels in growing chickens due to its influence on the activity of the T3-degrading type III deiodinase. The effectiveness of exogenous GH administration to acutely increase plasma T3 probably depends on the balance between the injected dose and the endogenous GH concentration, the hepatic GH receptor availability and the hepatic type III deiodinase level.

Effects of insulin-like growth factor-I (IGF-I) infusion and dietary tri-iodothyronine (T3) supplementation on growth, body composition and plasma hormone levels in sex-linked dwarf mutant and normal chickens.

This study used a sex-linked dwarf mutant (SLD) chicken to evaluate growth-promoting and metabolic effects of recombinant human insulin-like growth factor-I (rhIGF-I) treatment. The SLD chicken is characterized by a 30% reduction in body weight and by high plasma GH levels, low plasma IGF-I and triodothyronine (T3) levels and very low GH binding on liver membranes, suggesting reduced functional GH receptors compared with normal chickens. The effects of a continuous s.c. infusion by osmotic mini-pump of 0.1 mg rhIGF-I/kg per day from 4 to 8 weeks of age on body weight, bone growth and body composition were investigated in female SLD and normal chicks. In addition, half of the birds received a dietary supplement of T3 (0.1 parts per million). Plasma levels of IGF-I, GH, T3, thyroxine and insulin were followed during the treatment. In normal chicks, rhIGF-I infusion had no effect on growth and little effect on plasma hormone levels except for a decrease in plasma insulin. In dwarf chicks, rhIGF-I infusion slightly increased body weight but had no effect on longitudinal bone growth. In addition, plasma GH levels were decreased and T3 levels remained lower than in normal chicks. Normal and dwarf chicks showed a decrease in abdominal fat after both IGF-I administration and T3 supplementation, the treatments having additive effects in dwarf chicks only. The combined rhIGF-I and T3 treatment restored a quasi-normal hormonal pattern in dwarf chicks, except for insulin which remained lower than in normal chicks. These results suggest that IGF-I in the chicken has no direct endocrine effect on statural growth.

Selenium content of livers from sex-linked dwarf and normal broiler breeders. Influence of a thyrotropin-releasing hormone-induced growth hormone release.

Plasma concentrations of cGH, T3, and T4 were not different between dwarf and normal broiler breeders. Normal hens had a liver selenium content of 710 +/- 35 ng/g, and dwarf hens 656 +/- nine ng/g (n = 8). Following injections into a wing vein of different doses (1.5, 3, 6, 12, and 24 micrograms/kg) of the hypothalamic hormone TRH, GH was increased after 15 min. This effect seemed to last longer in dwarf chickens. Plasma concentrations of T3 increased significantly 1 h after TRH in normal hens, but TRH was ineffective in raising T3 levels in dwarf animals. The selenium content of livers obtained following decapitation after 2 h was also increased in normal hens up to 902 +/- 42 ng/g using the highest dose of TRH (24 micrograms/kg). This seemed not to be the case for dwarf animals. A much smaller number of hepatic cGH receptors was also found in dwarf hens, whereas the affinity of the hepatic GH receptor was not influenced by the genotype. It is concluded that the sex-linked dwarf hens are unable to increase their hepatic T4 into T3 conversion following a TRH challenge probably because of a deficiency in hepatic GH receptors. The lower content of selenium in dwarfs and their inability to increase its uptake after TRH seem therefore to support the hypothesis that selenium has a direct role in the activity of the 5'-deiodinase complex.

Effect of recombinant human insulin-like growth factor-I on weight gain, fat content, and hormonal parameters in broiler chickens.

Osmotic minipumps were implanted in 4-wk-old female broiler chickens to supply a 2-wk continuous infusion of recombinant human insulin-like growth factor-I (rhIGF-I) in three doses (.03, .1, and .3 mg/kg BW per day). At the end of the experimental period no differences in BW were detected, although abdominal fat was significantly reduced in the highest dose group. Measurement of fat content in both breast and thigh muscle indicated a different effect of IGF-I treatment on these parameters, as no reduction was observed. Determination of circulating IGF-I levels revealed a twofold increase in the .3-mg group whereas the lowest dose did not increase circulating plasma levels. The changes in IGF-I levels did not influence growth hormone levels whereas thyroxine levels were significantly decreased both in the .03- and .3-mg groups after 1 wk of treatment. At the same time plasma triiodothyronine levels were increased in the .1- and .3-mg/kg groups. These results indicate that a continuous infusion of IGF-I did not increase weight gain but may play a role as a fat repartitioning agent.

Effect of triiodothyronine supplementation on thyrotropin-releasing hormone-induced growth hormone secretion in sex-linked dwarf and normal chicks.

The effect of a dietary triiodothyronine (T3) supplement, of either 0.1 or 0.5 microgram/g of feed, was studied on the thyrotropin-releasing hormone (TRH)-induced growth hormone (GH) secretion in sex-linked dwarf (dw) or normal (Dw) chicks of both sexes. In normal chicks, 0.1 microgram/g T3 decreased plasma GH levels before TRH as well as the GH increase after TRH, and 0.5 microgram/g T3 totally suppressed any response to TRH, either at 4 or at 7 weeks of age. Dwarf chicks were more sensitive to TRH than normals when receiving either 0 or 0.1 microgram/g T3; 0.5 microgram/g T3 abolished the difference between genotypes at 4 weeks of age but not so clearly at 7 weeks of age, where dwarf females showed a slight but still significant GH increase after TRH. Interactions between genotype, TRH injection, and T3 treatments were often significant at 4 weeks of age and even more at 7 weeks of age. Dwarf chicks receiving 0.1 microgram/g T3, expected to have normal plasma T3 levels, showed a higher GH response after TRH. This suggests that other hormones may be involved in the regulation of this response, particularly IGF-I, which is known to remain at a low level in T3-treated dwarf chicks.

Food intake after hatching inhibits the growth hormone induced stimulation of the thyroxine to triiodothyronine conversion in the chicken.

The effect of a single injection of 10 micrograms chicken GH on circulating thyroid hormones as well as in vitro liver 5'-monodeiodination (5'-D) activity was studied in posthatch chicks submitted to different feeding conditions. One group was normally fed after hatching, a second group was only fed after three days and a third group was food deprived after 2 days of feeding. Combination of all results indicates that the start of food intake abolishes the stimulatory effect of a GH injection on circulating T3 and liver 5'-D activity. Food deprivation after a period of food intake restores the GH effect on plasma T3 but not on liver 5'-D.

Effect of hypophysectomy and acute administration of growth hormone (GH) on GH-receptor binding in chick liver membranes.

The effects of hypophysectomy on GH binding to liver membranes of young chicks were studied 3 days and 1 week after surgery. Specific binding of 125I-labelled chicken GH (cGH) to MgCl2-treated liver microsomal fractions of hypophysectomized animals was two- to fivefold greater than to those of sham-operated or control (non-operated) birds. This effect was due to a rise in binding capacity rather than a change in binding affinity of the GH receptor. Two daily injections of cGH (20 micrograms/animal) returned the number of hepatic GH receptors from hypophysectomized chicks to the level of the sham-operated ones. Administration of GH to the latter group did not cause a significant lowering of specific binding or number of receptors. No positive correlation between GH binding and plasma concentrations of insulin-like growth factor-I (IGF-I) was observed; although GH binding increased, IGF-I levels were lower for the hypophysectomized group. Since the number of hepatic GH receptors and the plasma GH levels were inversely correlated, it was concluded that the GH receptors in the liver of the chicken can be down-regulated by GH. This possibly explains why GH binding is low in posthatch and young chicks, because circulating GH concentrations are high during this period.

Differences in hepatic growth hormone receptor binding during development of normal and dwarf chickens.

The aim of this study was to compare the growth hormone (GH) receptor in liver microsomal fractions of normal chickens (Dw) and chickens carrying the dwarf gene (dw). Specific binding of GH to its hepatic receptor was significantly higher for Dw embryos from d 14 till d 20 of incubation than for dw embryos. The difference in binding was due to a decreased binding capacity but not affinity in the livers of the dwarf embryos. The same binding pattern was found in livers of adult chickens: lower binding was again caused by a lower number of GH receptors and at this stage the difference was even clearer than during embryonic development. Binding studies on livers of growing chicks demonstrated that binding was low for both genotypes, but a small though significant difference between them remained. The cause of this decrease in number of GH receptors in dwarf birds has yet to be determined but may be due to the primary action of the dwarf gene.

Effects of dietary T3 on growth parameters and hormone levels in normal and sex-linked dwarf chickens.

Tri-iodothyronine (T3) has been administered in the diet, from day of hatch until 8 weeks of age, to sex-linked dwarf and normal chicks of both sexes from a brown-egg slow-growing strain. Feed was supplemented with either 0.1 ppm or 0.5 ppm T3. A significant genotype by treatment interaction was observed on body weight: the effect of T3 in males was significantly positive for dwarfs and null for normals, the effect in females was null for dwarfs and significantly negative for normals. Feed efficiency was rather decreased by the treatment in both genotypes. Abdominal fatness was decreased in a dose-dependent manner in both genotypes, while rectal temperature was raised in dwarf chicks only. Plasma T3 was raised to normal levels in dwarfs receiving 0.1 ppm exogenous T3, while the 0.5 ppm dose yielded hyperthyroid levels. Plasma GH levels were decreased in a dose-dependent manner by the T3 treatment, the effect being larger in dwarfs. Surprisingly, plasma IGF-I was unchanged in spite of the GH decrease, whatever the genotype or the sex. It was concluded that exogenous T3 alone can have a stimulatory effect on growth in dwarfs but can not fully restore a normal growth rate. Both T3 and IGF-I are important for a normal growth and the relationship between T3 and IGF-I production should be further investigated in order to better understand the physiological modifications due to the sex-linked dwarf gene.

Ontogeny of the effect of purified chicken growth hormone on the liver 5' monodeiodination activity in the chicken: reversal of the activity after hatching.

The ontogeny of the effect of chicken growth hormone (c-GH) on the liver 5' monodeiodination (5'-D) activity was studied in chickens starting from 14-day-old embryos until 5-week-old chickens. Ten micrograms of the purified hormone was injected intravenously and after 2 hr, blood and liver samples were taken. In all embryonic stages tested, c-GH stimulated the peripheral thyroxine (T4) to triiodothyronine (T3) conversion as demonstrated by the increased plasma T3 level and liver 5'-D activity while the plasma reverse T3 (rT3) level was decreased. In chicks 1 day after hatching, GH was still able to increase the plasma T3 level and the liver 5'-D activity. However, in 2-day-old chicks, this stimulation had completely disappeared, while the control value of the plasma T3/T4 ratio was considerably higher than the day before. The effect of c-GH injection gradually turned into a decrease of 5'-D activity and plasma T3 in 5- and 7-day-old chicks. Further on, no effect of c-GH could be found in 2- to 4-week-old growing chickens, but in 5-week-old GH-injected animals the plasma T4 and rT3 level decreased again, although no significant effect on T3 or liver 5'-D activity could be demonstrated.

Impaired peripheral T3 production but normal induced thyroid hormone secretion in the sex-linked dwarf chick embryo.

Plasma concentrations of thyroxine (T4), triiodothyronine (T3) and chicken GH (cGH), together with hepatic 5'-monodeiodination (5'-D) activity, were measured in normal (Dw) and dwarf chick (dw) embryos at incubation d 18. An injection of 10 micrograms of ovine GH (oGH) raised plasma concentrations of T3 in Dw embryos after 1 and 2 h and stimulated hepatic 5'-D activity after 2 h. A non-specific increase in T4 was also observed after 1 h in Dw animals probably due to the heterologous nature of the injection. These effects were not observed in dw embryos. An injection of 1 microgram of TRH was able to increase cGH levels after 15 min in Dw embryos, whereas the the observed increase in the dw group was not significant. In Dw embryos, 0.01, 0.1 and 1 microgram of TRH increased plasma concentrations of T3 in a dose-dependent way, whereas in dw embryos, no reaction to the TRH injections was seen, except for the highest dose used. Contrary to this observation, T4 was increased to the same level in both Dw and dw embryos following TRH injections. An injection of 1 microgram of ovine CRH increased corticosterone after 0.5 h and elevated T3 and T4 after 2 h to the same extent in Dw and dw embryos. It is concluded that the thyrotrophic activities of TRH and oCRH and the corticotropic activity of oCRH do not differ between normal and sex-linked dwarf embryos. However TRH and GH were unable to stimulate the T4-T3 conversion in the liver of dw embryos, presumably due to the lack of hepatic GH receptors in these animals.

A decreased capacity of hepatic growth hormone (GH) receptors and failure of thyrotrophin-releasing hormone to stimulate the peripheral conversion of thyroxine into triiodothyronine in sex-linked dwarf broiler hens.

The effect of two different doses of thyrotrophic releasing hormone (TRH) upon the plasma levels of growth (GH) and thyroid hormones in both sex-linked dwarf (dw) and normal (Dw) broiler hens was determined. In normal hens, 1.5 and 24 microg TRH/kg increased the GH plasma concentrations after 15 min. Plasma concentrations of T3 increased significantly 1 h after TRH injection, whereas T4 concentration decreased after 2 following injection of 24 microg/kg TRH. In dwarf hens both doses of TRH increased the plasma concentrations of GH and the GH response lasted longer. However, TRH was ineffective in raising T3 and T4 levels. Saline-injected dwarf birds showed no differences in plasma T4 and T3 levels in comparison with normal hens. A smaller number of hepatic cGH receptors was found in dwarf hens, whereas the affinity of the hepatic GH receptor was not influenced by the genotype. It is concluded that the sex-linked dwarf broiler hen is unable to respond to a TRH-induced GH stimulus probably because of a deficiency in hepatic GH receptors resulting in a failure to stimulate the T4 to T3 converting activity.

Evidence for chicken GH as the only hypophyseal factor responsible for the stimulation of hepatic 5'-monodeiodination activity in the chick embryo.

The influence of an intravenous injection of chicken growth hormone (cGH), a total chicken pars distalis (PD) extract, and a PD extract depleted of cGH by immunoadsorption was studied in the 18-d-old chick embryo. Plasma concentrations of triiodothyronine (T3), thyroxine (T4), and hepatic 5'-monodeiodination (5'-D) activity were measured. An injection of total PD extract raised plasma T3, T4, and 5'-D activity, whereas a PD extract depleted of GH only increased plasma T4. The amount of cGH present in the PD extracts, as measured by homologous cGH radioimmunoassay, increased T3 and raised liver 5'-D, but had no effect on plasma T4. The effect on liver 5'-D was more pronounced with cGH than with a total PD extract, whereas the effect on plasma T3 was somewhat less pronounced. It was concluded that cGH increased the peripheral conversion of T4 into T3 in the chick embryo, whereas a PD extract depleted of cGH was purely thyrotropic. The PD extract also seemed to have 5'-D-suppressing activity.

Effect of the sex-linked dwarf gene on thyrotrophic and somatotrophic axes in the chick embryo.

Plasma concentrations of thyroxine (T4), triiodothyronine (T3), reversed triiodothyronine (rT3), and insulin-like growth factors I and II (IGF-I, IGF-II) together with peripheral 5'-monodeiodination activity were measured in both normal and sex-linked dwarf embryos between day 14 of incubation and day 1 posthatch. Plasma T4 levels increased gradually during embryonic development while T3 concentrations remained low until day 20, when a sharp increase was observed. rT3 levels also increased from day 14 and dropped on day 20 when T3 levels started to increase. 5'-monodeiodination activity was high on day 14 of incubation, decreased thereafter, and showed an increase at the time of air sac penetration together with increased T3 levels. At this stage, differences between normal and dwarf embryos were observed; the latter had lower nonsignificant 5'-Monodeiodination activity and lower (P less than 0.01) plasma T3 levels. Plasma IGF-II levels were high during the whole embryonic period studied. Dwarf embryos had lower (P less than 0.05) IGF-II levels at the time of hatching. IGF-I levels were high on days 14 and 16, declined afterwards, and started to increase again around hatching. With the exception of T3 and IGF-II levels, introduction of the dwarf gene did not cause major changes in the hormonal parameters studied. This may explain the identical body weight at hatching.

Hypothalamic hormones that release growth hormone stimulate hepatic 5'-monodeiodination activity in the chick embryo.

Plasma GH, tri-iodothyronine (T3), thyroxine (T4) and liver 5'-monodeiodination (5'-D) activity were measured in 18-day-old chick embryos injected with thyrotrophin-releasing hormone (TRH) and human pancreatic growth hormone releasing factor (hpGRF). Injections of 0.1 and 1 microgram TRH and 1.5 micrograms hpGRF increased the concentration of plasma GH while injection of 15 micrograms hpGRF had no effect. Concentrations of plasma T3 were raised after injection of TRH or hpGRF. Injections of TRH but not of hpGRF raised the concentration of plasma T4. The increases in concentration of plasma T3 after injection of TRH or hpGRF were parallelled by increases in liver 5'-D activity. An injection of 0.25 micrograms T4 significantly raised the concentration of T4 in plasma but had no effect on plasma T3 or liver 5'-D activity. It is concluded that the release of chicken GH by TRH or hpGRF is responsible for the observed increase in plasma concentration of T3 and liver 5'-D activity.