Age-related changes in acute central leptin effects on energy balance are promoted by obesity.

Leptin is a key catabolic regulator of food intake (FI) and energy expenditure. Both aging and obesity have been shown to induce leptin-resistance. The present study aimed to analyze age-related changes in the anorexigenic and hypermetabolic responsiveness to acute intracerebroventricular leptin administration in different age-groups of normally fed male Wistar rats (adult and old rats from 3 to 24months of age, NF3 to NF24, respectively). The expressions of the long form of the leptin receptor (Ob-Rb) and inhibitory SOCS3 genes were also assessed by quantitative RT-PCR in the arcuate nucleus (ARC). The influence of high-fat diet-induced obesity (HF) on the anorexigenic leptin effects were also tested in younger and older middle-aged groups (HF6 and HF12). Leptin-induced anorexia varied with age: leptin suppressed re-feeding FI (following 48-h fasting) strongly in young adult (NF3), but not in younger or older middle-aged (NF6 or NF12) or in aging (NF18) rats. However, anorexigenic leptin effects reached statistical significance again in old NF24 rats. Leptin-induced hypermetabolism, on the other hand, showed monotonous age-related decline and disappeared by old age. Ob-Rb expression declined until 12months of age followed by a partial recovery in NF18 and NF24 groups. On the other hand, SOCS3 expression was high in NF6 and NF18 and to some extent in NF24 rats. Age-related alterations of Ob-Rb and SOCS3 expression in the ARC may partly contribute to the explanation of age-related variations in anorexigenic but not hypermetabolic leptin effects. High-fat diet-induced obesity was associated with resistance to leptin-induced anorexia in HF6, similar to that seen in NF6. However, instead of the expected leptin-resistance in HF12, a strong leptin-induced suppression of re-feeding was detected in these obese middle-aged rats. Our results suggest that acute central effects of leptin on anorexia and hypermetabolism change in disparate ways during aging, implying separate mechanisms (e.g. signal transduction pathways) of different leptin actions. The age-related pattern shown by leptin-induced anorexia may contribute to the explanation of middle-aged obesity, and partly to that of aging anorexia. Our findings concerning obese rats are in accord with previous observations on anorexigenic effects of peripherally administered cholecystokinin: diet-induced obesity appeared to accelerate the development of age-related regulatory alterations. Similarly, our present data also raise the possibility that chronic diet-induced obesity promotes responsiveness to centrally applied leptin at least concerning anorexigenic effects.

Age-related alterations in the central thermoregulatory responsiveness to alpha-MSH.

Alpha-melanocyte-stimulating-hormone (alpha-MSH) is a neuropeptide that induces weight loss via its anorexigenic and hypermetabolic/hyperthermic effects. Two major public health problems of the human population involving energy balance (i.e. middle-aged obesity and aging cachexia) also appear in other mammals, therefore age-related regulatory alterations may also be assumed in the background. Previous studies demonstrated characteristic age-related shifts in the anorexigenic effects of centrally applied alpha-MSH with strong effects in young adult, diminished efficacy in middle-aged and very pronounced responsiveness in old rats. The present study aimed to investigate age-related changes in the acute central thermoregulatory responsiveness to an alpha-MSH injection in rats and to compare them with those of food intake-related responsiveness. Oxygen consumption (VO2), core (Tc) and tail skin temperatures (Ts, indicating heat loss) of male Wistar rats of different age groups (from 2 to 24 months of age), were recorded in an indirect calorimeter complemented by thermocouples upon intracerebroventricular alpha-MSH administration (0, 5 µg) at a slightly subthermoneutral environment (25-26 °C). Acute alpha-MSH-induced rises in VO2 and Tc were most pronounced in the young adult age-group. In these rats the hyperthemic effects were somewhat diminished by an activation of heat loss. Juvenile animals showed weaker hyperthermic responses, middle-aged rats none at all. Alpha-MSH-induced hyperthermia became significant again in old rats. Acute thermoregulatory (hypermetabolic/hyperthermic) responsiveness to alpha-MSH shows a distinct age-related pattern similar to that of acute anorexigenic responsiveness. Thus, our results may also contribute to the explanation of both middle-aged obesity and aging cachexia.

Bioassay-comparison of the antioxidant efficacy of hydrogen sulfide and superoxide dismutase in isolated arteries and veins.

Recent studies suggest that hydrogen sulfide (H2S) exhibits potent antioxidant capacity and improves vascular and tissue functions. Thus we aimed to compare the antioxidant efficacy of H2S to that of superoxide dismutase (SOD).Isometric force of isolated rat carotid arteries and gracilis veins was measured with a myograph. The vasomotor effect of the superoxide-generator pyrogallol (10-5M) was obtained in control conditions, and then in the presence of SOD (120 U/ml) or H2S (10-5M or 10-4M), respectively. Spectrophotometric measurements were performed to detect the effect of SOD and H2S on the auto-oxidation of pyrogallol.Pyrogallol increased the isometric force of carotid arteries (9.7 ± 0.8 mN), which was abolished by SOD (5.3 ± 0.8 mN), was not affected by 10-5M H2S (9.1 ± 0.5 mN), whereas 10-4M H2S slightly, but significantly reduced it (8.1 ± 0.7 mN). Pyrogallol significantly increased the isometric force of gracilis veins (1.3 ± 0.2 mN), which was abolished by SOD (0.9 ± 0.2 mN), whereas 10-5M (1.3 ± 0.2 mN), or 10-4M H2S (1.2 ± 0.2 mN) did not affect it. Pyrogallol-induced superoxide production was measured by a spectrophotometer (A420 = 0.19 ± 0.0). SOD reduced absorbance (A420 = 0.02 ± 0.0), whereas 10-5M H2S did not (A420 = 0.18 ± 0.0) and 10-4M H2S slightly reduced it (A420 = 0.15 ± 0.0).These data suggest that H2S is a less effective vascular antioxidant than SOD. We propose that the previously described beneficial effects of H2S are unlikely to be related to its direct effect on superoxide.

Effects of repeated surgical stress on daily changes of body core temperature in mice.

Daily body core temperature rhythm has been known to become blunted for several days following intra-abdominal implantation of biotelemetry transmitters in small rodents and about a week is required for re-establishment of stable body core temperature oscillation. In the present study carried out on mice it was found that a repetition of the same minor surgical intervention (laparotomy) several days apart could speed up the stabilization of body temperature oscillations. Melatonin supplied with the drinking water continuously was found to speed up the return of stable daily body temperature rhythm further on consecutive laparotomies, while daily injections of methylprednisolone resulted in some delay in the development of stable body core temperature oscillations. It is concluded that in C57BL/6 mice possessing low plasma levels of melatonin exhibit an adaptive response to repeated stresses influencing the dynamics of daily body temperature rhythm.

Energetics of fasting heterothermia in TRPV1-KO and wild type mice.

To learn the possible role of TRPV1 in the changes of temperature regulation induced by short-term energy lack, TRPV1-KO and wild type mice were exposed to complete fasting for 2 or 3 days while their core temperature and locomotor activity were recorded using a biotelemetry method. In both types of mice, fasting led to progressive daytime hypothermia with night-time core temperature being maintained at normothermia (collectively called heterothermia). During fasting rises of locomotor activity were observed parallel to night-time normothermia with occasional increases of both parameters recorded every 2 to 3 hours (ultradian rhythms). The daytime fall of core temperature was significantly greater in wild type than in TRPV1-KO mice, in the former an advance of the temperature/activity rhythm having been observed in spite of the presence of a 12/12 hour light/darkness schedule. Re-feeding applied at the beginning of the light-period led to rapid reappearance of normothermia in both types of mice without a large increase in locomotor activity. It is concluded that the TRPV1-gene may have a role in the development of adaptive daytime hypothermia (and hence saving some energy) in mice during complete fasting but still allowing normothermia maintained at night, a strategy probably serving survival under natural conditions in small size rodents such as the mouse. The possible role of muscle thermogenesis either with or without gross bodily movement during fasting or on re-feeding, respectively, may be based on different mechanisms yet to be clarified.

Pregnancies associated with low maternal serum alpha-fetoprotein concentrations.

PIP: A program of routine screening for antenatal detection of neural tube defects by alpha fetoprotein (AFP) testing is described. It was decided to determine the outcome of all pregnancies in which the patients exhibited a low AFP level during the screening process. From May 1975 to September 1976, 3% (53 cases) of the 2100 women tested had low AFP levels. Since maternal serum AFP levels rise with increasing gestational age until 32 weeks, a large number of women with low AFP levels actually have gestational ages more advanced than originally thought. That was true in this test. Ultrasonic examination of the cases with low AFP levels halped in revision gestational ages for some of the women. Among the 49 women with low AFP levels at gestational ages from 12-16 weeks, 12% had a spontaneous abortion eventually, 2% (or 1 case) had a macerated stillborn infant, and 1 woman was not pregnant. All 4 of the women with low AFP levels at gestational ages above 16 weeks had their gestational ages revised downward following ultrasonic examination. The study show that most women with low AFP levels eventually deliver a normal infant; a low AFP level does appear to be associated with spontaneous abortion. A larger study might identify association with other abnormalities. The study data was graphed and tabulated.^ieng

Antenatal screening in Oxford for fetal neural tube defects.

Between May 1975 and the end of 1977, 6443 antenatal patients were screened mainly between 16 and 22 weeks of pregnancy for neural tube defects (NTDs) at the John Radcliffe Hospital, Oxford, by maternal serum alpha-fetoprotein (AFP) measurement; a take-up of 72 per cent. Seventeen out of 18 (94 per cent) patients with open NTD pregnancies (9 out of 9 with anencephaly and 8 out of 9 with open spina bifida) had positive screening tests, and all except one were offered and accepted a termination of pregnancy. Two hundred and forty-five (3.8 per cent) patients with unaffected pregnancies also had positive screening tests, although only 1.4 per cent had an amniocentesis. Following ultrasonography, about 50 per cent of patients with unaffected pregnancies with positive screening tests were not offered an amniocentesis because they had a multiple pregnancy or their gestational age had been underestimated. The odds of having a fetus with an NTD among the women who had an amniocentesis was about 1 to 6 (1 to 11 for open spina bifida alone). Two apparently normal pregnancies were terminated. A survey of the acceptability of the screening programme among a consecutive sample of 73 patients who knew that they had a positive screening test revealed that all except one had no objection to screening in general, and 68 (93 per cent) wanted to be tested again in a future pregnancy. The approximate direct cost of the programme was 2 pounds to 3 pounds per patient screened, or about 1000 pounds per NTD detected (about 2200 pounds per open spina bifida detected).