Measurement properties of the 6-min walk test in individuals with exercise-induced pulmonary arterial hypertension.

BACKGROUND: Exercise-induced pulmonary arterial hypertension (EIPAH) is associated with reduced peak exercise cardiac output (CO) and aerobic capacity (peak ). We investigated the validity of the encouraged 6-min walk test (6MWT) to identify exercise limitation and estimate aerobic capacity in subjects with EIPAH. METHODS: Seventeen subjects with EIPAH (56 ± 14 years, 15 women) and 20 healthy controls (57 ± 13 years, 19 women) underwent two encouraged 6MWTs and a symptom-limited cardiopulmonary exercise test (CPET). To measure central haemodynamics, subjects with EIPAH performed the CPET with a pulmonary artery catheter in situ. RESULTS: Compared with controls, subjects with EIPAH had reduced peak (1.2 ± 0.4 vs 1.7 ± 0.5, L/min, P < 0.01), 6-min walk distance (6MWD) (575 ± 86 vs 669 ± 76 m, P < 0.001) and 6-min walk work (6MWW) (39 ± 11 vs 45 ± 7 km.kg, P < 0.01). In subjects with EIPAH, there was a moderate correlation between 6MWD and peak (r= 0.72, P < 0.01) and a strong correlation between 6MWW and peak (r= 0.86, P < 0.001). There were significant correlations between 6MWD and peak CO (r= 0.59, P < 0.05), and between peak and peak CO (r= 0.55, P < 0.05). Peak heart rate was similar in the CPET and 6MWT in subjects with EIPAH (133 ± 15 vs 133 ± 19 beats/min, P= 0.8). CONCLUSIONS: The encouraged 6MWT identifies reduced exercise capacity and provides a valid estimate of aerobic capacity in EIPAH.

The effects of carbon dioxide on exercise-induced asthma: an unlikely explanation for the effects of Buteyko breathing training.

OBJECTIVES: To examine the effect of breathing 3% CO2 on exercise-induced asthma (EIA), as a raised airway CO2 level is suggested to mediate the effects of Buteyko breathing training (BBT). DESIGN: Double-blind crossover study, using a standard laboratory-based exercise challenge, with EIA defined as a fall of 15% or greater in the forced expiratory volume in one second (FEV1) within 30 minutes of completing a standard exercise protocol. SUBJECTS: 10 adults with confirmed EIA. INTERVENTION: Air enriched with 3% CO2 during and for 10 minutes after exercise. OUTCOME MEASURES: Maximum percentage fall in FEV1 after exercise. Area under curve (AUC) of the decrease in FEV1 with time. RESULTS: Mean maximum fall in FEV1 was similar: 19.9% with air, and 26.9% with 3% CO2 (P = 0.12). The mean AUC for the total 30-minute post-exercise period was 355 for air and 520 for 3% CO2 (P = 0.07). After discontinuing the 3% CO2 at 10 minutes after exercise, there was a further and sustained fall in FEV1. Mean AUC for the period 10-30 minutes post-exercise was significantly greater for CO2 than air (275 and 137, respectively [P = 0.02]). Mean minute ventilation was increased when subjects exercised breathing 3% CO2: 77.5 L/min for 3% CO2, compared with 68.7 L/min for air (P = 0.02). CONCLUSION: Breathing 3% CO2 during exercise does not prevent EIA. The shape of the FEV1 response curve after 3% CO2 suggests that a greater degree of EIA (because of increased minute ventilation during exercise) was opposed by a direct relaxant effect of CO2 on the airway. Increased airway CO2 alone is an unlikely mechanism for the reported benefits of BBT; nevertheless, further study of the effects of voluntary hypoventilation in asthma is warranted.

Relationship between liver glycogen metabolism and glucose homeostasis in the fetal rat: studies with the gsd/gsd rat.

Litters containing both heterozygous and homozygous gsd/gsd rats have been studied to determine whether fetal liver glycogen contributes to the fetal blood glucose pool. It is shown that homozygous gsd/gsd fetuses (unable to mobilise liver glycogen) have consistently lower blood glucose concentrations than their heterozygous (normal) littermates at all ages studied. This difference is independent of maternal type and maternal blood glucose concentration. There is a marked activation of liver phosphorylase in heterozygous fetuses at 21 days of gestation, associated with a significant rise in the fetal/maternal blood glucose ratio. In contrast, there is no significant activation of phosphorylase or rise in the ratio for homozygous littermates. In homozygous fetuses, the blood glucose concentrations remained low and liver glycogen high. The possibility of fetal liver glycogen being derived from gluconeogenic precursors is discussed.

Effect of changes in litter size at mid-lactation on lactation in rats.

The effect of altering the number of pups per litter from ten to two or two to ten at day 14 of lactation in rats was investigated. Reducing litter size had no effect on the daily live weight gain whereas increasing the litter size resulted in an initial weight loss followed by an impaired weight gain. Maternal food consumption decreased to values appropriate for rats feeding two pups within 2 days after litter reduction, but required at least 5 days after the increase in litter size to increase to values appropriate for rats feeding large litters from parturition. Altering the litter size resulted in maternal serum insulin concentrations that were intermediate between those expected for rats feeding two or ten pups from parturition. Maternal serum prolactin concentrations increased after litter sizes were increased and decreased initially after litter reduction before being restored to normal levels. The activities of the lipogenic enzymes, fatty acid synthase, glucose-6-phosphate dehydrogenase and 'malic' enzyme in the mammary gland all decreased within 72 h of litter reduction to levels appropriate for rats feeding two pups. Although all three enzymes increased in activity after litter size increase, only fatty acid synthase had reached values appropriate for rats feeding ten pups by 72 h.

Insulin in the rat fetus. A new form of circulating insulin.

The apparent hyperinsulinemia seen in the fetal rat in late gestation (day 20) is shown to be due to an immunoreactive insulin (IRI) species peculiar to the fetal rat, and not monomeric insulin. This species disappears within 12 h after birth and only insulin as found in the adult can then be detected. The fetal serum IRI has a molecular weight of approximately 11,000 and differs from monomeric insulin and proinsulin in its behavior on gel filtration and anion-exchange chromatography. The activity of fetal serum in an insulin bioassay based on the incorporation of glucose into lipid was half that expected on the basis of the serum IRI concentration. The addition of fetal serum to purified insulin or proinsulin caused an increase in the apparent molecular weight of both hormones by 2000. In the case of insulin, the generated species of mol wt 8000 was clearly differentiated from the endogenous species (11,000) on Sephadex G50. The fetal pancreas at this age was found to contain predominantly monomeric insulin that had full biologic activity in the insulin bioassay. It is postulated that the fetal pancreas secretes monomeric insulin, which is then modified in the circulation to a complex with reduced biologic activity. This process appears to have two steps in that in vitro fetal serum increases the apparent molecular weight of added insulin to 8000, whereas in vivo an additional component of approximately 3000 mol wt is involved to produce the endogenous insulin complex.

Effect of chronic consumption of a high-fat diet on mammary metabolism.

Rats have been fed semi-synthetic diets containing 4 and 20% (w/w) corn oil for periods of 9-12 weeks covering two lactations. The mean maternal and pup weights at day-14 of the second lactation on the diets were not significantly different. The milk fatty acids of the rats on the high-fat diet contained 39 mol% linoleic acid and only 20 mol% medium-chain acids compared with over 40% for the low-fat diet. With the exception of "malic" enzyme, none of five enzymes assayed in mammary supernatants was significantly altered by the diet fed. These results suggest that the synthesis of these enzymes in the mammary gland is insensitive to dietary lipid.

The effect of starvation and refeeding on lipogenic enzymes in mammary glands and livers of lactating rats.

Lactating rats were starved for 48 h and refed a high-carbohydrate diet for a further 48 h. Starvation stops milk secretion, which resumes shortly after refeeding. Three lipogenic enzymes, fatty acid synthase, glucose 6-phosphate dehydrogenase (EC 1.1.1.49) and 'malic' enzyme (EC 1.1.1.40) all decrease in the mammary gland during starvation and are restored to the pre-starvation levels 48 h after refeeding. The same enzymes in liver also decrease during starvation, but increase to values significantly higher than those for the normal fed rats after refeeding the high-carbohydrate diet. For the fatty acid synthase these values were four times the pre-starvation values. Serum insulin and prolactin concentrations also increased upon refeeding the high-carbohydrate diet.

Evidence for insulin degradation by muscle and fat tissue in an insulin resistant diabetic patient.

The period before, during and after resistance to subcutaneous insulin in a 20-year-old, non-obese insulin-dependent patient with 'brittle' diabetes is documented and clinical observations are correlated with experimental results of insulin degradation in vitro. Treatment with intravenous but not subcutaneous aprotinin markedly reduced subcutaneous insulin requirements. Insulin resistance recurred following cessation of aprotinin. Serum free insulin levels were low during the subcutaneous resistant phase compared with those during the more sensitive phase. Insulin degradation in vitro by adipose tissue and muscle taken from the patient during a resistant phase was increased compared with degradation by tissue taken during a sensitive phase and by tissue from normal subjects. Chromatography of incubation media revealed that during the resistant phase, tissue from the patient degraded insulin to small fragments. It is concluded that, in this patient, insulin resistance was caused by excessive degradation in both muscle and adipose tissue.

Glucose homeostasis during the perinatal period in normal rats and rats with a glycogen storage disorder.

The fetal rat mobilizes liver glycogen during parturition for use as a glucose source until the onset of gluconeogenesis at 2 h after birth. A rat strain (NZR/Mh) unable to mobilize liver glycogen because of a phosphorylase b kinase deficiency has been used to assess the importance of liver glycogen in glucose homeostasis of the newborn. In normal rats the mean blood glucose concentration of the fetus measured at various times up to 24 h after natural birth ranged between 3.7 and 5.4 mM. In contrast, fetuses of the affected rats were hypoglycemic before birth (2.02 +/- 0.15 mM), and by 1 h after birth the blood glucose had decreased to 0.74 +/- 0.14 mM. Concentrations increased by 4 h to 1.48 +/- 0.17 mM and by 24 h reached values not significantly different from the normal newborn rats. Changes in plasma insulin over the perinatal period were similar in both groups although concentrations were always significantly lower in the affected rts. The findings demonstrate the crucial role of the fetal liver glycogen store in the maintenance of normoglycemia in the newborn. The normal rat does not develop hypoglycemia when born naturally and left with the mother after birth (in contrast to other studies in which the newborn were taken by cesarian delivery 1 d prematurely and kept in an artificial environment without food). The rats with the glycogen storage disorder experienced severe hypoglycemia without any apparent effects, which raises questions concerning alternative fuels available to and utilized by the newborn.

Liver glycogen synthase in the developing foetal rat.

Kinetic constants for liver glycogen synthase (UDPglucose: glycogen 4-alpha-D-glucosyltransferase, EC 2.4.1.11) with respect to UDPglucose have been measured in foetal liver homogenates from samples taken during late gestation (days 17-22) and the first hours after birth. The V of the inactive form of glycogen synthase increased markedly in this period and there was a significant increase in V of the active enzyme to a maximum at day 20 of gestation. The Km for UDPglucose measured in the presence of glucose-6-P (total activity) did not vary greatly, mean values of 0.51 +/- 0.04 mM. Values derived for the inactive enzyme were almost identical. In contrast, Km values for active glycogen synthase in foetal livers during gestation were significantly higher than those for adult liver. Highest values were seen at day 19 of gestation (1.84 +/- 0.08 mM) followed by a steady fall to 0.55 +/- 0.05 mM in the newborn compared with a mean value of 0.48 +/- 0.04 mM for adult liver. Existence of a reduced affinity of active glycogen synthase for UDPglucose must be recognized when assaying the enzyme in foetal liver, particularly when extrapolating values to rates of glycogen synthesis in vivo. Data were obtained only after removal of an amylase-like contaminant from foetal liver samples which invalidated the radioassay of glycogen synthase. This work illustrates the care needed in the analysis of foetal tissue and the interpretation of resulting data when utilizing methods developed for adult tissue.