Delafossite structure of heterogenite polytypes (HCoO2) by Raman and infrared micro-spectroscopy.

Heterogenite is commonly referred in mineralogy literature as a cobalt oxy-hydroxide CoO(OH). However, detailed analysis of Raman and infrared spectra acquired on particularly well-crystallized natural samples of heterogenite suggests that the mineral can be characterized by a delafossite-type structure, with a general chemical formula ABO2. Indeed, the Raman spectrum of heterogenite, along the one with grimaldiite (HCrO2), lacks visible free OH-group vibrational modes, while the infrared spectrum shows strong hydrogen bond absorption bands. HCoO2 is thus a better formulation of heterogenite that describes more clearly its vibrational behavior and avoids the confusion in literature. Electronic backscattered diffraction (EBSD) is then used to distinguish and map the 2H and 3R heterogenite natural polytypes for the first time. The comparison of EBSD and Raman mappings clearly indicates that the 2H polytype is characterized by an additional peak at 1220 cm(-1). The presence/absence is therefore an efficient tool to distinguish both polytypes.

Raman spectroscopy as a tool to characterize heterogenite (CoO·OH) (Katanga Province, Democratic Republic of Congo).

Natural heterogenite (CoO·OH) samples were studied by Raman microspectroscopy, electronic microprobe and Electronic BackScattered Diffraction (EBSD). Raw samples and polished sections were made from 10 mines covering the Katanga copperbelt (Katanga Province, Democratic Republic of Congo). Four typical Raman responses have been obtained leading to investigate the laser-induced dehydroxylation of heterogenite into a Co-spinel structure. The results are also compared with EBSD patterns from oven heated heterogenite samples. A close relationship was established between the chemical substitutions of Co by mainly Cu, Ni, Mn and Al and their impact on the mineral Raman response.

Galanin receptor antagonists decrease fat preference in Brattleboro rat.

The Brattleboro rat eats spontaneously 46% of its diet per day in fat when given a choice of carbohydrate, protein and fat. An overexpression of galanin (GAL) has been also observed in the hypothalamic paraventricular nuclei (PVN). This associative correlation has led to a hypothesis of a functional relation between central galanin expression and the preference for a lipid diet. In the present experiments, the effects of two GAL receptor antagonists, C7 and galantide, on fat consumption and central overexpression of GAL were investigated. Both antagonists were injected into either the cerebral ventricles or directly above the PVN, and the diet consumption followed for the subsequent 24h. C7 decreased significantly fat consumption when injected into the ventricles or directly above the PVN. In contrast, galantide must be injected above the PVN to show the same effect. However, the two antagonists did not modify GAL mRNA expression in the PVN when they were injected 2h before sacrifice. These experiments confirm a functional link between the preferential consumption of fat and hypothalamic Galanin; different subtypes of the GAL receptor are probably involved, since both Galanin antagonists were differently efficient in decreasing spontaneous fat selection of the Brattleboro rat.

Plasma leptin and hypothalamic neuropeptide Y and galanin levels in Long-Evans rats with marked dietary preferences.

Neuropeptides present in the hypothalamus and new messengers in the periphery such as leptin modulate food intake in mammals. Neuropeptide Y (NPY) and galanin in microdissected brain areas and plasma leptin levels were measured by specific radioimmunoassays during the resting period in rats selected for their strong preference either for carbohydrate or fat, but with identical energy intake. NPY concentrations were 23% lower (p <.02) in carbohydrate-preferring (CP) than in fat-preferring (FP) rats in the parvocellular part of the paraventricular nucleus (PVN), which is one of the main areas involved in the regulation of feeding behavior. On the other hand, galanin was significantly (+25%, p = .03) higher in CP rats than in FP rats in the magnocellular part of the PVN. Plasma leptin was more than 50% higher in FP rats than in CP rats (p < .01) and highly correlated with the fat preference (r = 0.57; p = .003) and body weight gain. We conclude that the rats with a spontaneous and marked dietary preference have a characteristic peptidergic profile. Due to their anatomical relationships, neuropeptide Y could act in conjunction with galanin in a peptidergic balance located in the paraventricular nucleus. This model integrates information provided by the energy stores and translated by peripheral messengers such as leptin which could act in a counterregulatory manner in order to limit the overweight induced by the ingestion of unbalanced diets.

Effect of chronic intraperitoneal injections of leptin on hypothalamic neurotensin content and food intake.

This study was intended for the investigation of the effects of chronic injections of leptin for 7 days on food intake and hypothalamic neurotensin (NT). Leptin treatment significantly reduced food intake [144.3+/-2.5 g (L) vs. 156.7+/-2.5 g (C); P=0. 002] and body weight gain [23.7 g+/-1.0 g (L) vs. 31.5+/-1.3 g (C); P=0.003]. NT concentration was lower in the lateral hypothalamus (LH) of leptin-treated rats than in the control ad libitum fed rats (-30%; P<0.05). The same diminution was observed in pair-fed rats (-27%; P<0.05). This diminution was therefore related to the decrease in food intake rather than to a direct effect of leptin. As the LH was the only area where NT was modified, it appears that among the hypothalamic nuclei involved in the regulation of feeding behavior it is the most sensitive area to a low energy depletion. Therefore, it might play a specific role in triggering the mechanisms necessary to restore body weight and/or energy balance.

Consequences of inspired oxygen fraction manipulation on myocardial oxygen pressure, adenosine and lactate concentrations: a combined myocardial microdialysis and sensitive oxygen electrode study in pigs.

Adenosine is a potent vasodilator whose concentration has been shown to increase in cardiac tissue in response to hypoxia. However, the time-dependent relationship between the levels of myocardial interstitial adenosine and tissue oxygenation has not yet been completely established. Therefore, the purpose of this study was to investigate the complex relationship between tissue myocardial oxygen tension (PtiO(2)) and interstitial myocardial adenosine and lactate concentrations by developing a new technique which combines a cardiac microdialysis probe and a Clark-type P O(2)electrode. The combined and the single microdialysis probes were implanted in the left ventricular myocardium of anesthetized pigs. The consequences of the combined use of microdialysis and P O(2)probes on myocardial PtiO(2)and microdialysis performances against glucose were evaluated. A moderate but significant reduction in the relative recovery against glucose of the combined probe was observed when compared to that of the single microdialysis probe (42+/-2 v 32+/-1%, mean+/-S.E. M.n=5 P<0.05), at 2microl/min microdialysis probe perfusion flow. Similarly, myocardial oxygen enrichment, measured by the P O(2)electrode, was negligible when microdialysis probe perfusion flow was 2microl/min. Systemic hypoxia (FiO(2)=0.08) resulted in a significant decrease in PtiO(2)from 30+/-4 to 11+/-2 mmHg, limited increase in coronary blood flow (CBF), and a significant increase in myocardial adenosine and lactate concentrations from 0.34+/-0.05 to 0.98+/-0.06micromol/l and from 0.45+/-0.05 to 0.97+/-0.06 mmol/l respectively (P<0.05). Increasing the FiO(2)to 0.3 restored the PtiO(2)and hemodynamic parameters to baseline values with no changes in interstitial adenosine and lactate concentrations. Nevertheless, myocardial interstitial adenosine remained significantly higher than baseline values. In conclusion, this study demonstrates the ability of a combined probe to measure simultaneously regional myocardial PtiO(2)and metabolite concentration during hypoxia. The hypoxia-induced increase in myocardial adenosine persists after correction of hypoxia. The physiological significance of this observation requires further studies.

Consequences of labetalol administration on myocardial beta adrenergic receptors in the brain dead pig.

OBJECTIVES: Cardiac dysfunction following brain death is associated with highly increased myocardial norepinephrine, lactate and adenosine concentrations. Administration of labetalol, a mixed alpha-, beta-adrenergic receptor antagonist, attenuates metabolic disturbances and improves myocardial function. The purpose of this study was to investigate beta-adrenergic receptor (beta AR) density and affinity in the presence or absence of labetalol administration, as a possible mechanism of the protective effects of this drug. METHODS: Experimental animals were divided into three groups: sham-operated, brain-dead pigs, and brain-dead pigs treated with labetalol (10 +/- 3 mg/kg). The maximum number of binding sites (Bmax) and the dissociation constant (Kd) of beta AR were determined with (-)-[125I]cyanopindolol on myocardial samples harvested 3 hours after brain death. RESULTS: Left ventricular beta AR density and affinity were identical in brain-dead and sham-operated animals. Labetalol-treated pigs exhibited a significant decrease of Bmax and an increase of Kd as compared with brain-dead pigs. Bmax decrease was due to the persistence of labetalol in the membrane preparations. Increased Kd was too low to be biologically significant. Therefore, beta AR number and affinity can be considered as unchanged after adrenergic blockade with labetalol. CONCLUSIONS: The protective mechanism of labetalol on brain death-induced myocardial dysfunction cannot be explained by changes in beta AR density and affinity but is probably related to a preservation of the oxygen consumption/oxygen delivery balance during the autonomic storm.

Dietary composition during fetal and neonatal life affects neuropeptide Y functioning in adult offspring.

The aim of this study was to examine the impact of maternal diet during the gestation and lactation periods on the neuropeptide Y (NPY) system in adult offspring. Male Long-Evans rats were obtained from dams fed either on a well-balanced diet (C), a high carbohydrate diet (HC) or a high-fat diet (HF) and fed themselves on the well-balanced diet for their whole life. At 6 months of age, their feeding response to various doses of NPY injected in the lateral brain ventricle was measured in one group and NPY concentrations in microdissected nuclei of the hypothalamic were measured in a second group. The HF rats were lighter than the two other groups (P<0.001). The control rats showed a typical dose-dependent feeding response to NPY. The HC rats showed a continuous increase in the response, starting at the intermediate dose (1.0 microg) only while the HF rats had a maximal response at the lowest dose (0.5 microg). The HF rats ate twice as much as the HC rats at the lowest dose tested 1 h after injection (4.4+/-0.6 vs. 2.7+/-0.4 g; P<0.05), showing therefore the greatest sensitivity to NPY. This change in the sensitivity was not related to hypothalamic NPY concentration as it was not modified in the arcuate and paraventricular nuclei. The diet imposed on the mother could have long-lasting effects on body weight regulation of the offsprings and alter the NPY system likely through modifications at the receptor level.

Dietary preferences of Brattleboro rats correlated with an overexpression of galanin in the hypothalamus.

Galanin (GAL) is a neuropeptide cosynthesized with vasopressin (AVP) in neurons of the hypothalamo-neurohypophysial system. It increases food intake when injected into the brain and elicits an overconsumption of fat. The Brattleboro rat (DI) is genetically unable to produce AVP; the AVP-deficient-producing neurons of the hypothalamo-neurohypophysial system of DI rats are chronically stimulated and DI rats suffer from diabetes insipidus. We studied the central expression of GAL and the dietary preferences in the DI rat. GAL was overexpressed in the hypothalamus of the DI rat. GAL mRNA was higher by 1.8-fold in the supraoptic (P < 0.05) and by four-fold in the paraventricular nuclei (P < 0.001) of male and female DI rats compared with those of control Long Evans (LE) rats. However, GAL mRNA was lower in the arcuate nuclei of DI rats and equal to that of LE rats in the dorsomedian nuclei. We also measured a high preference for a lipid diet (45% of the daily consumption) when DI rats ate from a choice of the three macronutrients. Chronic infusion with deamino-8D-AVP (agonist of AVP V2 receptors) prevented the diabetes insipidus and the chronic stimulation of the hypothalamo-neurohypophysial system of the DI rats. However, the treatment did not suppress the overexpression of GAL, nor did it affect the rats' preference for a lipid diet. We conclude that the DI rat provides a novel animal model in which a spontaneous dietary preference correlates with the overexpression of one of the hypothalamic peptides, GAL.

Increase in myocardial interstitial adenosine and net lactate production in brain-dead pigs: an in vivo microdialysis study.

BACKGROUND: Brain death-related cardiovascular dysfunction has been documented; however, its mechanisms remain poorly understood. We investigated changes in myocardial function and metabolism in brain-dead and control pigs. METHODS: Heart rate, systolic (SAP) and mean (MAP) arterial pressure, left ventricular (LV) dP/dtmax, rate-pressure product, cardiac output (CO), left anterior descending coronary artery blood flow, lactate metabolism, and interstitial myocardial purine metabolite concentrations, monitored by cardiac microdialysis, were studied. A volume expansion protocol was performed at the end of the study. RESULTS: After brain death, a transient increase in heart rate (from 90 [67-120] to 158 [120-200] beats/min) (median, with range in brackets), MAP (82 [74-103] to 117 [85-142] mmHg), LV dP/dtmax (1750 [1100-2100] to 5150 [4000-62,000] mmHg x sec(-1), rate-pressure product (9100 [7700-9700] beats mmHg/min to 22,750 [20,000-26,000] beats mmHg/min), CO (2.2 [2.0-4.0] to 3.3 [3.0-6.0] L/min), and a limited increase in left anterior descending coronary artery blood flow (40 [30-60] to 72 [50-85] ml/min) were observed. Net myocardial lactate production occurred (27 [4-40] to -22 [-28, -11] mg/L, P<0.05) and persisted for 2 hr. A 6-7-fold increase in adenosine dialysate concentration was observed after brain death induction (2.9 [1.0-5.8] to 15.8 [7.0-50.7] micromol/L), followed by a slow decline. Volume expansion significantly increased MAP, CO, and LV dP/dtmax in control animals, but decreased LV dP/dtmax and slightly increased CO in brain-dead animals. A significant increase in adenosine concentration was observed in both groups, with higher levels (P<0.05) in brain-dead animals. CONCLUSIONS: Brain death increased oxygen demand in the presence of a limited increase in coronary blood flow, resulting in net myocardial lactate production and increased interstitial adenosine concentration consistent with an imbalance between myocardial oxygen demand and supply. This may have contributed to the early impairment of cardiac function in brain-dead animals revealed by rapid volume infusion.

Evidence that hypothalamic neurotensin signals leptin effects on feeding behavior in normal and fat-preferring rats.

Leptin inhibits food intake when it is injected in the periphery or in the central nervous system. It is likely that its action is not only mediated by the inhibition of orexigenic peptides such as neuropeptide Y. Therefore, we characterized the pharmacological and physiological relationships of leptin with neurotensin (NT), a central feeding inhibitor. Firstly, we investigated the central interactions of leptin and NT. Intracerebro-ventricular (ICV) injection were done in normal Long-Evans rats. NT had a short lasting (30 minutes; p<0.01) inhibitory effect on spontaneous food intake measured at the beginning of the dark phase whereas the effect of ICV leptin was observed after 24 hours (p<0.001). Co-injection with leptin potentiated NT effect at 30 minutes (p<0. 001) and prolonged it for 30 additional minutes (p<0.01). In addition, NT potentiated the effect of leptin at 30 and 60 minutes (p<0.02 and p<0.001 respectively) but not at 24 hours. Secondly, we observed that NT concentrations were augmented in selective brain areas in fat-preferring rats (+ 34% for hypothalamic NT; p<0.03). This increase was observed in the parvocellular part of the paraventricular nucleus (PVNp) only and was associated with an increase in circulating leptin levels (+ 75%; p<0.003). Interestingly, plasma leptin and NT in the PVNp were strongly correlated (r=0.57; p<0.003), suggesting changes of NT processing or release in this nucleus. These results strongly suggest that the short-term anorexigenic effects of leptin in normal rats are at least partly mediated by changes in NT processing or release. They also suggest that these processes take place in the hypothalamus, most probably in the PVNp and that they might be sensitive to fat ingestion. Therefore, the neurotensin increase observed in fat-preferring rats would limit the overconsumption of energy, a physiological mechanism translated by leptin.

Hypothalamic neuropeptide Y content and mRNA expression in weanling rats subjected to dietary manipulations during fetal and neonatal life.

Hypothalamic neuropeptide Y (NPY) is present very early during the fetal life and is rapidly functional in the regulation of feeding behavior after birth. In the present experiment, we tried to determine the influence that the diet type ingested by dams during gestation and lactation would have on the growth and hypothalamic and pancreatic peptides of their progeny immediately after weaning. The dams were fed on either a high-carbohydrate (HC), a high-fat (HF) or a control diet ad libitum. At 3 days of age, the HC pups weighed significantly more than the two other groups (P < 0.02 vs. C and P < 0.002 vs. HF). At weaning, the HF rats were significantly lighter than the two other groups (P < 0.001). Food intake was significantly lower in the HF rats than in the two other groups 3 days (P < 0.002) and 5 days after weaning (P < 0.02). Plasma glucose of the HF rats was significantly lower than that of the control rats (P < 0.05) and of the HC rats (P < 0.01). Immunoreactive insulin in the HF rats was also significantly lower than that in the control rats (-53%; P < 0.001) and in the HC rats (-47%; P < 0.001). NPY content and mRNA expression in the arcuate nucleus were not significantly different between the three groups. NPY concentration only varied in the ventromedian nucleus. In the control rats, it was significantly lower than that of the HC rats (-35%; P < 0.01) and that of the HF rats (-32%; P < 0.002). These data demonstrated that the regulatory mechanisms of feeding behavior in offspring are completely and differentially modified by the macronutrient content of the diets ingested by their mother. Both peripheral and central mediators were strongly implicated. These modifications could have long-term repercussions on body weight and composition.

Behavioral deficits in monosodium glutamate rats: specific changes in the structure of feeding behavior.

We studied the feeding rhythms and feeding patterns of adult Long-Evans rats treated with monosodium glutamate (MSG) in their early post-natal period. This treatment is known to induce neuronal degeneration in the arcuate nucleus (ARC), a major hypothalamic site implicated in the regulation of feeding. Neonatal rats were treated intraperitoneally with MSG or saline (controls) alone on the first days of life. At age of 6 months, male control and male MSG rats were placed in our automatic feeding system, and the structure of feeding behavior and diurnal feeding rhythms were analysed. On a 24 hours basis, MSG rats ate less than control rats (-24%). This hypophagia resulted from a mild diurnal hyperphagia (+6%) and a pronounced nocturnal hypophagia (-34%). This hypophagia was the main consequence of a decrease of meal size in MSG rats (-37%) and was associated with an increase in meal duration (+52%). It was also associated with a total disappearance of the two feeding peaks that normally occur at light and dark onset in the rat (-90% 2 h after dark onset and -49% 2 h before light onset). These results indicate that neonatal treatment with MSG induces important changes in feeding patterns and feeding rhythms in the adulthood. These changes might be related to the disappearance of neurotransmitters located in the arcuate nucleus.

Neuropeptide Y release in the paraventricular nucleus of Long-Evans rats treated with leptin.

Neuropeptide Y (NPY) and leptin are actually two of the most potent peptides involved in the regulation of food intake with their respective stimulatory and inhibitory actions. The infusion of each peptide has a significant influence on the mRNA expression of the other in the adipose tissue for leptin and in the arcuate nucleus of the hypothalamus for NPY. To confirm this functional interaction, we measured the in vivo release of hypothalamic neuropeptide Y in awake fasted and refed Long-Evans rats after intraperitoneal (I.P.) injection of leptin. For this purpose, we used the push-pull perfusion technique with the cannula placed above the right paraventricular nucleus. I.P. leptin significantly inhibited food intake during the two hours of food access (-50%; p < 0.02). NPY release was not modified by leptin alone. But, when food was present, it slightly but significantly increased (p < 0.03 or less) and remained at a sustained level in the leptin-treated rats whereas it decreased in the control saline-injected group (p < 0.04). Thus, leptin did not acutely regulate NPY release and other food-related factors are probably involved as mediators of its anorexigenic effects.

Hypothalamic neuropeptide Y and plasma leptin after long-term high-fat feeding in the rat.

The ingestion of fat by rodents affects the level of neuropeptide Y (NPY) in the hypothalamus and we hypothesized that they might be linked via leptin, the adipose tissue hormone. The influence of fat intake on leptin and NPY levels was studied in rats fed on either a high-fat (HF) or a low fat diet (LF) for 5 months. Ingestion of the HF diet increased fat deposition (+48%; P < 0.01), leptinemia (+189%; P < 0.001) and reduced NPY levels in the arcuate nucleus (-35%; P < 0.01) and in the paraventricular nucleus (-22%; P < 0.01). However, although leptin levels reflected the amount of relative fat deposition (r = 0.62; P < 0.01), we found no evidence for a direct relationship between plasma leptin and NPY levels in the hypothalamus. These results suggest that the long-term effects of fat intake on NPY concentrations in the hypothalamus and plasma leptin are associated with different regulatory mechanisms.

Neuropeptide Y in the Ventromedial and Suprachiasmatic Nuclei: Role in the Feeding Pattern of Monosodium Glutamate-Treated Rats.

Neuropeptide Y (NPY) is an important mediator for the light/dark cycle entrainment of circadian rhythms including feeding rhythm. It is present in the main circadian oscillator, the suprachiasmatic nucleus which receives NPY efferences from the intergeniculate leaflet to form a pathway named the geniculo-hypothalamic tract (GHT). In the present experiment, we measured NPY in the arcuate nucleus (ARC) and in areas more or less related to feeding rhythm regulation in rats with brain lesion induced by monosodium glutamate (MSG). Newborn Long-Evans rats were subcutaneously injected three times during the first week of life either with MSG (4 mg/g BW; n = 15) or with saline (n = 10). One week after weaning, food intake and NPY concentrations in the GHT, ventromedial (VMN) and dorsomedial nuclei and lateral hypothalamus were measured. Daily food intake in the MSG-treated rats was slightly lower than in control (saline) rats (10.8 ± 0.3 vs 11.9 ± 0.4 g; p < 0.03). This diminution in food intake was only observed during the dark period (p < 0.02). NPY in the ARC rats was 50% lower in MSG-treated rats than in control rats (p < 0.001) confirming therefore the neurotoxicity of the neonatal injections of high doses of MSG. A smaller decrease was observed in the VMN of MSG-treated rats (-30%; p < 0.02) whereas no variations were observed in the GHT and in the other areas. We conclude that when the GHT is intact, gross regulation of feeding rhythm is maintained. The VMN might contribute to the slight change in the dark/light distribution of food intake through the NPY variation.

Arginine vasopressin (AVP) depletion in neurons of the suprachiasmatic nuclei affects the AVP content of the paraventricular neurons and stimulates adrenocorticotrophic hormone release.

Arginine vasopressin (AVP) produced in the hypothalamic suprachiasmatic nuclei (SCN) plays a role in establishing neuroendocrine rhythms and, in particular, in regulating the corticotrope axis rhythm. It has recently been shown that AVP from SCN inhibits corticosteroid release. In order to investigate the influence of suprachiasmatic AVP on the different peptidergic systems through the hypothalamus, SCN neurons containing AVP were functionally lesioned by using toxins associated with a cytotoxic monoclonal antibody (MAb) raised against AVP. Six days later, the AVP contents and AVP mRNA were measured in different hypothalamic and extrahypothalamic sites. Adrenocorticotrophic hormone (ACTH) concentration was also measured in plasma. Microinjection of the AVP-MAb/toxin mixture into SCN brought about a significant decrease in the AVP expression in SCN. This is demonstrated by the decrease in the AVP immunoreactive content (24%, P < 0.01) and the decrease of AVP hybridized mRNA (33%, P < 0.01). This points to the efficiency of the microinjection in decreasing the production of AVP in the injection area. Modifications of the AVP contents in the two subdivisions of the hypothalamic paraventricular nucleus (PVN) were also observed. AVP contents decreased in the parvocellular subdivision (pPVN); this is coherent with the AVP depletion in SCN since pPVN is the major site of the SCN hypothalamic efferences. AVP content and AVP mRNA increased in the magnocellular subdivision (mPVN); this also confirms the difference in AVP synthesis regulation according to the PVN subdivisions. The microinjection did not modify AVP expression in supraoptic nuclei or oxytocin (OT) immunoreactive content in the main hypothalamic OT containing sites. Plasma ACTH values were double (P < 0.02) the values measured under non-specific IgG treatment 10 hr after lights on. This probably resulted from the stimulation of the hypothalamo-pituitary-adrenal system since corticotrophin-releasing hormone (CRH) mRNA increased simultaneously by 24% (P < 0.05) in the PVN and the immunoreactive CRH content of the median eminence significantly decreased (26%, P < 0.05). Overall, our data confirm that AVP produced in the SCN inhibits the CRH-adrenocorticotrope axis in normal conditions, probably because of SCN projections of AVP neurons on the PVN.