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Cytokine production and lymphocyte subpopulations in aged humans. An assessment during nocturnal sleep.

The view of a general impairment of immune functions associated with aging has been challenged by recent studies including a more detailed evaluation of various cytokines and lymphocyte subsets. In the present human study, effects of age on the production of cytokines by T cells and monocytes were assessed, together with age-dependent changes in subset populations of mononuclear cells (MNC). Blood was collected every 30 min during nocturnal sleep in 16 aged (mean: 79.6 +/- 7.5 years) and in 16 young controls (mean: 24.6 +/- 3.1 years). Nocturnal sleep was chosen as a well-defined period within the 24-h cycle with minimal exogenous influences. The in vitro production of interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) was measured after mitogen stimulation with lipopolisaccharide from E. coli (LPS). Production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) was measured after stimulation with phytohemagglutinin (PHA). Regarding MNC subsets, monocytes, lymphocytes, CD3+, CD4+, CD8+, HLA-DR, CD16+, CD25+, and CD19+ were determined. Advanced age was associated with a decreased number of T cells (CD3+) and decreases in the major T cell subsets (CD4+, CD8+, P < 0.001). Production of IL-2 was not affected. However, production of IFN-gamma tended to be enhanced, and numbers of activated T cells (HLA-DR/CD3+), natural killer cells (CD16+), and T cells expressing IL-2 receptors (CD25+/CD3+) were markedly increased in the aged. While monocyte counts were unchanged in the elderly production of IL-1 beta and TNF-alpha mainly derived from these cells, was enhanced (p < 0.05). Results indicate a state of enhanced responsiveness of the T cell compartment and of monocytes in aged which may compensate for the substantial decrease in T cells.

Low cerebrospinal fluid insulin levels in obese humans

To the Editor: In healthy men, insulin concentrations in cerebrospinal fluid (CSF) increase after acute elevations of plasma insulin levels, indicating that the brain content of insulin is normally finely tuned to circulating insulin [1]. Central nervous insulin promotes weight loss [2] and attenuates peripheral insulin resistance via inhibition of hepatic gluconeogenesis [3]. With increasing body weight, plasma insulin levels rise as a result of elevated insulin resistance. It is not yet known whether in hyperinsulinaemic obese subjects this increase is accompanied by parallel increases in CSF insulin levels that would act to promote weight loss in these patients. We compared insulin concentrations in plasma and CSF in a group of 45 subjects (23 women, age 19–80 years) with a wide body weight range (BMI 16.24–38.10 kg/m). Five subjects were newly diagnosed as diabetic by elevated fasting plasma glucose levels (>7.0 mmol/l). Since their exclusion did not alter the results, their data were included in the analyses. After an overnight fast, subjects reported to the laboratory for simultaneous sampling of blood and CSF (1 ml) via lumbar puncture after local anaesthesia (2 ml mepivacain-HCl 1%). BMI and waist-to-hip ratio were assessed, and body composition was measured by standard bioelectrical impedance analysis (BIA 2000-M; Data Input, Frankfurt, Germany). Insulin was determined using a commercial competitive double-antibody RIA (Pharmacia Insulin RIA 100; Pharmacia Diagnostics, Upsalla, Sweden). To increase the sensitivity of the insulin assay for the expected lower concentrations in CSF, the standard procedure was slightly modified. In brief, insulin was determined using 100 μl of CSF, 50 μl of [I]insulin diluted with buffer at a ratio of 1:3, and 50 μl of insulin antiserum diluted at a ratio of 1:2. The incubation time was increased from 2 h in the standard method to 3 h at room temperature. With this procedure, threshold sensitivity was reliably lowered to 1.8 pmol/l. Intra-assay variation was <4.5% for determination in CSF and plasma. Insulin plasma and CSF concentrations were measured in duplicate and quadruplicate, respectively. The lowest standard used was 1.8 pmol/l, which could be reliably measured with <4.5% variation. Therefore, all measures below this standard were set to 1.8 pmol/l for analysis. Thus, the actual CSF:plasma insulin ratio may even be slightly lower than reported, although analyses using the measurements below the lowest standard did not alter the results. Additionally, plasma and CSF glucose concentrations were measured (Beckman glucose analyser II; Beckman Instruments, Fullerton, CA; coefficient of variation <1.1%). Insulin resistance was estimated according to the homeostasis model assessment (HOMA) formula, with high scores denoting a high degree of insulin resistance. Bivariate correlation coefficients were determined and a stepwise multiple regression analysis was performed to detect associations Diabetologia (2006) 49:2790–2792 DOI 10.1007/s00125-006-0409-y

Selective influence of the menstrual cycle on perception of stimuli with reproductive significance: An event‐related potential study

In this study, we examined changes in the event-related potential (ERP) to stimuli with and without reproductive significance occurring during the menstrual cycle. Eleven spontaneously cycling women were tested during three menstrual phases (menses, ovulatory phase, luteal phase) differing in plasma concentrations of gonadal hormones. ERPs were recorded while subjects were presented with slides showing pictures from four different stimulus categories (sexual stimuli, babies, people occupied with body care, ordinary people). Slides were presented randomly in the context of two tasks, requiring either affective processing (i.e., to judge the emotional content of a slide as positive, neutral, or negative) or structural processing (i.e., to estimate the number of parallel thin lines inserted in each picture). Menstrual phase primarily affected a late positive component (LPC) peaking 550-600 ms poststimulus. The effects were as follows: (i) During the ovulatory phase, amplitude of the LPC to sexual stimuli was larger than that evoked by the other stimulus categories. (ii) This relationship was not apparent during the other menstrual phases or (iii) during the ovulatory phase when the task required structural processing. The ovulatory increase in LPC positivity to sexual stimuli suggests a greater valence of these stimuli during a phase of increased sexual desire. The data indicate a specific effect of the menstrual cycle on the processing of sexual stimuli that increases with deeper emotional processing.

A regulatory role of prolactin, growth hormone, and corticosteroids for human T-cell production of cytokines.

The release of the pituitary hormones, prolactin and growth hormone (GH), and of adrenal corticosteroids is subject to a profound regulation by sleep. In addition these hormones are known to be involved in the regulation of the immune response. Here, we examined their role for in vitro production of T-cell cytokines. Specifically, we hypothesized that increased concentrations of prolactin and GH as well as a decrease in cortisol, i.e., hormonal changes characterizing early nocturnal sleep, could be responsible for a shift towards T helper 1 (Th1) cytokines during this time. Whole blood was sampled from 15 healthy humans in the morning after regular sleep and was activated in vitro with ionomycin and two concentrations of phorbol myrestate acetate (PMA, 8 and 25 ng/ml) in the absence or presence of prolactin, prolactin antibody, GH, glucocorticoid receptor (GR) antagonist RU-486, or mineralocorticoid receptor (MR) antagonist spironolactone. Hormones were examined at physiological concentrations. Production of T-cell derived cytokines was measured at the single cell level using multiparametric flow cytometry. Generally, effects were more pronounced after stimulation with 8 rather than 25 ng/ml PMA. The following changes reached significance (p <.05): prolactin (versus prolactin antibody) increased tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) producing CD4+ and CD8+ cells and interleukin-2 (IL-2) producing CD8+ cells. Compared with control, prolactin antibody decreased, whereas GH increased IFN-gamma+CD4+ cells. RU-486 increased TNF-alpha, IFN-gamma, and IL-2 producing CD4+ and CD8+ cells. Surprisingly strong effects were found after MR blocking with spironolactone which increased TNF-alpha, IFN-gamma, and IL-2 producing CD4+ and CD8+ cells. No effects on IL-4+CD4+ cells were observed, while the IFN-gamma/IL-4 ratio shifted towards Th1 after spironolactone and after RU-486 plus GH. Results suggest that enhanced prolactin and GH concentrations as well as low cortisol levels during early nocturnal sleep synergistically act to enhance Th1 cytokine activity.

Manipulating central nervous mechanisms of food intake and body weight regulation by intranasal administration of neuropeptides in man

Maintaining a stable body weight set-point is assumed to rely on a homeostatic central nervous system (CNS) regulation of body fat with the particular involvement of hypothalamic pathways. The peripheral adiposity signals insulin and leptin convey information on the amount of energy stored as body fat to the arcuate nucleus of the hypothalamus, where anabolic/orexigenic and catabolic/anorexigenic pathways interact to regulate food intake and energy expenditure. One of the most prominent orexigenic messengers is neuropeptide Y (NPY), whereas melanocortins, including alpha-melanocyte-stimulating hormone (alpha-MSH), are essential for inducing anorexigenic effects. The melanocortin receptor 4 (MC4-R) plays the most important role in mediating catabolic effects of alpha-MSH. In this review, we present a series of own studies on NPY, insulin and MSH/ACTH4-10, an MC4-R agonist. The studies were all based on the intranasal route of administration which enables a direct access of the peptides to hypothalamic functions. NPY acutely attenuated electrocortical signs of meal-related satiety. Prolonged intranasal administration of insulin as well as of MSH induced weight loss in healthy human subjects. However, overweight subjects did not lose body fat after MSH administration. The results corroborate in humans the significance of all three messengers for the central nervous regulation of adiposity and might contribute to the future development of medical strategies against body-weight-related disorders.

Chronic fentanyl application induces adrenocortical insufficiency

We report a case of a 64‐year‐old man with secondary adrenocortical insufficiency who has been on a chronic transdermal fentanyl treatment because of sciatic pain syndrome. Shortly before admission to our hospital, the patient had discontinued his hydrocortisone medication. Adrenal crisis was assumed and during therapy with hydrocortisone infusion, the patient recovered. We suspected an opiate‐induced suppression of the hypothalamus‐pituitary‐adrenal (HPA) axis. Therefore, we gradually reduced the opiate dosage. After 1 week, HPA axis function was markedly improved. We conclude that opiate medication may inhibit – in a life‐threatening way – the organisms ability to respond to physical, emotional or metabolic stressors.

To Dip or Not to Dip: On the Physiology of Blood Pressure Decrease During Nocturnal Sleep in Healthy Humans

That sleep is accompanied by a blood pressure decrease is well known; however, the underlying physiology deserves further investigation. The present study examines in healthy subjects 2 main questions: is this dipping actively evoked? and what are the consequences of nondipping for daytime blood pressure? Nocturnal blood pressure was extrinsically elevated in 12 sleeping subjects to mean daytime values by continuously infused phenylephrine. This nondipping significantly lowered morning blood pressure during rest and 3 hours after resuming physical activity compared with a control condition (isotonic saline). Neither muscle sympathetic nerve activity nor sensitivity of &agr;-adrenoceptors was reduced. However, the set point for initiation of regulatory responses through the baroreflex was clearly shifted toward lower blood pressure levels. Our results support the hypothesis of an actively regulated central mechanism for blood pressure resetting and set point consolidation of the baroreflex during nighttime sleep. This is suggested by the fact that extrinsically induced nondipping induces sustained decrease in blood pressure during the following morning through an actively lowered baroreflex set point.

Selective influence of menstrual cycle on perception of stimuli with reproductive significance.

&NA; Sixteen healthy women (20–32 years) not taking oral contraceptives were tested at three phases of their menstrual cycles (menses, preovulatory phase, and midluteal phase). Another 16 women taking oral contraceptives served as age‐matched controls, and were tested during menses and during phases corresponding to the preovulatory and midluteal phase. On each test occasion, perception was assessed of visual stimuli belonging to three categories: sex (nude men), babies, and stimuli related to body care. Also, meaningless syllables were given. Stimuli were presented by a tachistoscope. Presentation of each stimulus was repeated until the subject signaled recognition. Trials to recognition and correctness of the recognition response were recorded. In spontaneously cycling women effects of the phase of the menstrual cycle depended on the stimulus meaning: During the preovulatory phase, there was an increase in the number of correctly recognized sex stimuli (p < 0.05) and of stimuli falsely recognized as sex stimuli (p < 0.05). The number of stimuli falsely recognized as body care was reduced during this phase (p < 0.05). Moreover, during the luteal phase, the number of trials to recognition of baby stimuli was diminished (p < 0.01). In women taking contraceptives, no stimulus specific changes occurred, but correct recognition was generally improved during the luteal phase. Rather than supporting a general influence of the normal menstrual cycle on perceptual functions, the present results demonstrate that the effects of the menstrual cycle depend on the meaning of the stimulus with the perception of sexual stimuli, but not of stimuli related to maternal nurture, being improved during the preovulatory phase.

Towards the therapeutic use of intranasal neuropeptide administration in metabolic and cognitive disorders

The nose provides an effective way for delivering neuropeptides to the central nervous system, bypassing the blood-brain barrier and avoiding systemic side effects. Thereby intranasal neuropeptide administration enables the modulation of central nervous signaling pathways of body weight regulation and cognitive functions. Central nervous control of energy homeostasis is assumed to rely on hypothalamic neuropeptidergic pathways that are triggered by the peripheral adiposity signals insulin and leptin conveying the amount of body fat to the brain. Melanocortins, including alpha-melanocyte stimulating hormone (alpha-MSH), are essential for inducing anorexigenic/catabolic effects, i.e. for inhibiting caloric intake and increasing energy expenditure. Insulin, in addition to its function as an adiposity signal, also influences memory formation. Here we present a series of studies on the intranasal administration of MSH/ACTH4-10, a melanocortin receptor agonist, and of insulin. Prolonged administration of MSH/ACTH4-10 induced weight loss in normal-weight, but not in overweight humans. Intranasal insulin reduced body fat and improved memory functions in the absence of adverse peripheral side effects. Our results may contribute to the future development of therapeutic strategies in disorders like obesity and cognitive impairments that derive from dysfunctions of central nervous neuropeptidergic pathways.

Hyperinsulinemia causes activation of the hypothalamus-pituitary-adrenal axis in humans

OBJECTIVE: Hyperactivity of the hypothalamus-pituitary-adrenal (HPA) axis is frequently found in hyperinsulinemic subjects, such as patients with diabetes or abdominal obesity. Here, the question has been posed as to whether hyperinsulinemia increases HPA secretory activity.METHODS: We performed paired—euglycemic and stepwise hypoglycemic (76–66–56–46 mg/dl)—clamp experiments in two groups (each of 15 healthy men) at different insulin infusions rates, ie, 1.5 mU/min/kg (low-insulin condition) and 15.0 mU/min/kg (high-insulin condition).RESULTS: During the euglycemic clamp, the high rate insulin infusion increased plasma ACTH levels, whereas plasma ACTH levels remained essentially unchanged during the low-insulin condition (condition by time interaction, P=0.008). Likewise, serum cortisol levels were higher during the high- vs low-insulin condition (condition by time interaction, P=0.004). During the hypoglycemic clamp, plasma ACTH levels did not differ between the low- vs high-insulin condition, while serum cortisol levels were higher during the high- vs low-insulin condition at the beginning of the clamp (plasma glucose ∼76 mg/dl; P=0.032).CONCLUSION: Data indicate that hyperinsulinemia acutely increases HPA secretory activity in healthy men. This finding appears to be relevant to the pathogenesis of many clinical abnormalities associated which diabetes and abdominal adiposity, often referred to as the metabolic syndrome.