Blockade of central delta-opioid receptors inhibits salt appetite in sodium-depleted rats.

Various studies have investigated the role of central opioid peptides in feeding behavior; however, only a few have addressed the participation of opioids in the control of salt appetite. The present study investigated the effect of intracerebroventricular injections of the δ-opioid antagonist, naltrindole (5, 10 and 20 nmol/rat) and the agonist, deltorphin II (2.5, 5, 10 and 20 nmol/rat) on salt intake. Two protocols for inducing salt intake were used: sodium-depletion and the central injection of angiotensin II. In addition, the effect of a central δ-opioid receptor blockade on locomotor activity, on palatable solution intake (0.1% saccharin) and on blood pressure was also studied. The blockade of central δ-opioid receptors inhibits salt intake in sodium-depleted rats, while the pharmacological stimulation of these receptors increases salt intake in sodium-replete animals. Furthermore, the blockade of central δ-opioid receptors inhibits salt intake induced by central angiotensinergic stimulation. These data suggest that during sodium-depletion activation of the δ-opioid receptors regulates salt appetite to correct the sodium imbalance and it is possible that an interaction between opioidergic and angiotensinergic brain system participates in this control. Under normonatremic conditions, δ-opioid receptors may be necessary to modulate sodium intake, a response that could be mediated by angiotensin II. The decrease in salt intake following central δ-opioid receptors blockade does not appear to be due to a general inhibition of locomotor activity, changes in palatability or in blood pressure.

Central kappa opioid receptors modulate salt appetite in rats.

The role of the central opioid system in the control of water and salt intake is complex, with both stimulatory and inhibitory effects having been observed. The aim of the present study was to investigate the participation of the central κ-opioid receptors in the control of salt appetite. Male Wistar rats were submitted to two different experimental protocols: sodium deficit produced by the diuretic, furosemide, and brain angiotensinergic stimulation in rats under normal sodium balance. Lateral ventricle (LV) injections of Nor-binaltorphimine (Nor-BNI) at different doses (5, 10 and 20 nmol) inhibited hypertonic saline solution (1.5%) intake in sodium-depleted rats. The salt appetite induced by an LV injection of angiotensin II (Ang II) (10 ng) was also blocked by Nor-BNI injections into the LV, while no significant change was observed in water intake. Furthermore, the decrease in salt intake seems not to have been due to a general inhibition of locomotor activity or to any change in palatability, since central administration of Nor-BNI failed to modify the intake of a 0.1% saccharin solution when the animals were submitted to a "dessert test" or to induce any significant locomotor deficit in the open-field test. Also the central administration of Nor-BNI was unable to modify blood pressure in sodium-depleted animals. The present results suggest that activation of endogenous κ-opioid receptors modulates salt appetite induced by sodium depletion and by central angiotensinergic stimulation in rats.

Multiple opioid receptors mediate the hypotensive response induced by central 5-HT(3) receptor stimulation.

The aim of the present work was to investigate the role of brain µ, κ and δ opioid receptors in the central serotonergic mechanisms regulating blood pressure in rats. The data obtained show that: (1) pharmacological activation of central 5-HT(3) receptors yields a significant decrease in blood pressure; (2) the blockade of those receptors by a selective antagonist induces an acute hypertensive response; (3) the pharmacological blockade of central opioid receptors by three different opioid antagonists exhibiting variable degrees of selectivity to µ, κ and δ opioid receptors always suppressed the hypotensive response induced by central 5-HT(3) receptor stimulation; (4) the blockade of opioid receptors by the same opioid antagonists that impaired the hypotensive effect of central 5-HT(3) receptor stimulation failed to modify blood pressure in animals not submitted to pharmacological manipulations of central 5-HT(3) receptor function. It is shown that a 5-HT(3) receptor-dependent mechanism seems to be part of the brain serotonergic system that contributes to cardiovascular regulation since the hypertensive response observed after ondansetron administration indicates that central 5-HT(3) receptors exert a tonic inhibitory drive on blood pressure. Furthermore, the data obtained here clearly indicate that the hypotensive response observed after pharmacological stimulation of central 5-HT(3) receptors depends on the functional integrity of brain µ, κ and δ opioid receptors, suggesting that a functional interaction between serotonergic and opiatergic pathways in the brain is part of the complex, multifactorial system that regulates blood pressure in the central nervous system.

Box-Behnken design: an alternative for the optimization of analytical methods.

The present paper describes fundamentals, advantages and limitations of the Box-Behnken design (BBD) for the optimization of analytical methods. It establishes also a comparison between this design and composite central, three-level full factorial and Doehlert designs. A detailed study on factors and responses involved during the optimization of analytical systems is also presented. Functions developed for calculation of multiple responses are discussed, including the desirability function, which was proposed by Derringer and Suich in 1980. Concept and evaluation of robustness of analytical methods are also discussed. Finally, descriptions of applications of this technique for optimization of analytical methods are presented.

The central amygdala regulates sodium intake in sodium-depleted rats: role of 5-HT3 and 5-HT2C receptors.

In the present paper, we have evaluated the participation of 5-HT(3) and 5-HT(2C) receptors in the central amygdala (CeA) in the regulation of water and salt intake in sodium-depleted rats. m-CPBG-induced pharmacological activation of 5-HT(3) receptors located in the CeA resulted in a significant reduction in salt intake in sodium-depleted rats. This antinatriorexic effect of m-CPBG was reverted by pretreatment with the selective 5-HT(3) receptor antagonist ondansetron. The injection of ondansetron alone into the CeA had no effect on sodium-depleted and normonatremic rats. Conversely, pharmacological stimulation of 5-HT(2C) receptors located in the central amygdala by the selective 5-HT(2C) receptor agonist m-CPP failed to modify salt intake in sodium-depleted rats. Additionally, the administration of a selective 5-HT(2C) receptor blocker, SDZ SER 082, failed to modify salt intake in rats submitted to sodium depletion. These results lead to the conclusion that the pharmacological activation of 5-HT(3) receptors located within the CeA inhibits salt intake in sodium-depleted rats and that 5-HT(2C) receptors located within the CeA appear to be dissociated from the salt intake control mechanisms operating in the central amygdala.

Short stature, obesity and arterial hypertension in a very low income population in North-eastern Brazil.

BACKGROUND AND AIM: This cross-sectional study involved the adult population (age >18 and <60 years) of a 315-shack slum on the outskirts of the city of Maceió in North-eastern Brazil. The purpose was to investigate whether short stature in adults (an indicator of undernutrition in early life) is associated with arterial hypertension and obesity. METHODS AND RESULTS: We collected the subjects socio-economic data, and arterial hypertension (AH), weight, height, waist circumference and waist/hip (W/H) circumference ratio measurements. Hypertension was diagnosed as diastolic AH f 90 mmHg and/or systolic AH f 140 mmHg. The body mass index (BMI) was used to determine nutritional status, with overweight/obesity being defined on the basis of a cut-off point of 25 kg/m2. A W/H ratio of f 0.80 for women or f 0.95 for men was considered indicative of abdominal obesity. Short stature was defined as falling into the 1st quartile (Q) of height distribution. Hypertension was prevalent in 28.5% of the population (women=38.5%; men=18.4%). The systolic and diastolic AH readings were significantly higher in women in the 1st Q than in those in the 4th Q, and the same was true of W/H. The prevalence of hypertension was statistically significant for the first two Q's in comparison with the last two: 22.1% vs 14.6% (men), and 42.4% vs 34.6% (women). Hypertension was more prevalent in women who were obese and short (50%) than in those who were obese but not short (OR=1.98; CI=1.22-2.96). CONCLUSIONS: Living conditions were extremely precarious and the prevalence of hypertension was quite high. Stature negatively correlated with hypertension and overweight in women but not in men.

Food consumed does not account for the higher prevalence of obesity among stunted adults in a very-low-income population in the Northeast of Brazil (Maceió, Alagoas).

OBJECTIVE: To study the food pattern of stunted and nonstunted, obese and nonobese individuals in a very-low-income population. DESIGN: A household survey. SETTING: Slum set up by the 'Homeless Movement', city of Maceió (Alagoas), Brazil. SUBJECTS AND METHODS: A total of 532 adults classified by sex, stature (Z -2s.d. of the NCHS curves), and body mass index (BMI) were compared using the following variables: waist circumference, waist-hip circumference ratio (W/H), percentage body fat (skinfold thickness and bioelectrical impedance), and food intake (24-h recall). RESULTS: The prevalence of stunting was 22.6%. In all, 30% of the stunted subjects were overweight or obese, compared with 23% for the nonstunted individuals (P<0.05). In women, logistic regression analysis showed a strong association among weight, abdominal fat, and stunting (r=0.81). No significant differences were observed in the values of W/H or in the qualitative menu of the different categories. Energy intake was below the RDA figures (about 63%). There was similarity among the groups regarding the proportion of macronutrients, except for the fact that stunted obese women ingested less fat and protein than nonstunted obese women. Stunted obese individuals consumed less energy (5962 kJ) than the population as a whole (6213 kJ), an amount far lower than their average needs, which were calculated on the basis of their shorter stature (8109 kJ). CONCLUSION: The observed energy consumption seems compatible with the panorama of undernutrition present in the population, but it does not explain the high prevalence of obesity detected.

Mechanical isolation of capuchin monkey (Cebus apella) preantral ovarian follicles

The aim of this study was to adapt a mechanical procedure for the isolation of intact preantral follicles from Cebus apella ovaries. The interval effect of serial sections of the tissue chopper was tested on a number of preantral follicles isolated from ovaries (n=6) of three C. apella females, two prepubertal and one adult. Ovaries were divided into four equal parts and fragmented with a tissue chopper, adjusted for serial sections at intervals of 250, 500, 750 and 1,000æm, respectively. Isolated follicles were counted in a Neubauer's chamber and classified as primordial, primary or secondary. The number (mean±SE) of preantral follicles isolated from 1/4 ovary varied from 68,330+17,590 (at the 1,000æm cut interval) to 300,830+111,460 (at the 500æm cut interval. The mean diameter of the isolated preantral follicles varied from 11.6æm to 27.8æm.

Obesity and undernutrition in a very-low-income population in the city of Maceió, northeastern Brazil.

Obesity is the nutritional disorder which has shown the greatest increase in prevalence, even in those countries in which deficiency diseases represent a severe public health problem. The goal of the present study was to analyse the anthropometric profile of a community living in the outskirts of Maceió, capital of Alagoas (northeastern Brazil), and to investigate the hypothesis of a coexistence of undernutrition and obesity in a very low-income population. The survey was conducted on 315 families (1247 individuals). Among the children (aged < or =10 years), the prevalence of wasting, stunting and wasting plus stunting was 3.8, 8.3 and 8.7 % respectively. Wasting (10.2 %) was the most prevalent form of undernutrition among adolescents; nonetheless, a higher frequency of stunting (11 %) and overweight-obesity (5.5 %) was seen specifically in girls, in agreement with trends found in other studies. Adults exhibited a high prevalence of overweight-obesity (25 %), but stunting was also present (22 %). Of the stunted individuals, 30 % were overweight-obese and 16.3 % were underweight. There were eighty-six families with at least one parent who was underweight (27 %) and 104 families with at least one parent who was overweight (33 %). Underweight and overweight-obesity were both present in ninety-six households (30 %). These results may indicate that better living conditions in urban areas in a population 'adapted' to chronic famine might increase the susceptibility to obesity. Considering the harm caused by the cumulative effect of these two conditions (undernutrition in childhood and obesity in adult life) there is a clear need for new studies to uncover the determinant factors so that preventive measures can be implemented.

Molecular phylogeny of ateline new world monkeys (Platyrrhini, atelinae) based on gamma-globin gene sequences: evidence that brachyteles is the sister group of lagothrix.

Nucleotide sequences, each spanning approximately 7 kb of the contiguous gamma1 and gamma2 globin genomic loci, were determined for seven species representing all extant genera (Ateles, Lagothrix, Brachyteles, and Alouatta) of the New World monkey subfamily Atelinae. After aligning these seven ateline sequences with outgroup sequences from several other primate (non-ateline) genera, they were analyzed by maximum parsimony, maximum likelihood, and neighbor-joining algorithms. All three analyzes estimated the same phylogenetic relationships: [Alouatta [Ateles (Brachyteles, Lagothrix)]]. Brachyteles and Lagothrix are sister-groups supported by 100% of bootstrap replications in the parsimony analyses. Ateles joins this clade, followed by the basal genus Alouatta; these joinings were strongly supported, again with 100% bootstrap values. This cladistic pattern for the four ateline genera is congruent with that obtained in previous studies utilizing epsilon-globin, IRBP, and G6PD nuclear genomic sequences as well as mitochondrial COII sequences. Because the number of aligned nucleotide positions is much larger in the present datasetoff than in any of these other datasets, much stronger support was obtained for the cladistic classification that divides subfamily Atelinae into tribes Alouattini (Alouatta) and Atelini, while the latter divides into subtribes Atelina (Ateles) and Brachytelina (Brachyteles and Lagothrix).

Should nutrition be considered as a supplementary measure in schistosomiasis control?

The most important data on the relationships between nutritional status and schistosomiasis mansoni are reviewed. The probable impact of such findings on the traditional strategies for control of the disease are discussed. In endemic areas, malnutrition and schistosomiasis seem to be associated. Malnutrition impairs the biological development of the parasite. However, like the parasite, it also depresses the host's immune system, and malnutrition and infection can be mutually aggravating. Recent schistosomiasis-control activities, although apparently well designed, have frequently seemed ineffective because of the multiplicity of factors involved, and have not offered a realistic promise of sustainable and definitive control. However, these actions must be continued and even encouraged because they do lead to reductions in the prevalence of infection and, of particular importance, to reductions in the incidence of the more severe forms of the disease. Improvement of the nutritional status of those who inhabit endemic areas, particularly those on low incomes (who are at relatively high risk of malnutrition and of schistosomiasis), is recommended as a supplementary measure.

Electrophoretic polymorphisms and their taxonomic implications in Callitrichini (Primates, Platyrrhini).

Five hundred forty-three blood samples from 15 populations of the four genera of callitrichin primates were studied electrophoretically. Polymorphism and genetic distances were estimated for 20 loci, 13 of which were polymorphic. The lion tamarin (Leontopithecus) studied here exhibited the least variability for these loci, while the monospecific Cebuella showed the most. The genetic distances observed between Callithrix and Cebuella genera support previous evidence indicating a close taxonomic relationship between them. Genetic distance values obtained in this study also support the synonimyzation of the kuhli form with Callithrix jacchus penicillata.

A long-term intake of a protein hydrolysate seems to increase the risk of encephalopathy in mice infected with Schistosoma mansoni.

Previous investigations showed that Schistosoma mansoni infection aggravates protein malabsorption in undernourished mice and this can be reverted by administration of casein hydrolysate. The present study was undertaken to evaluate the effects of ingestion of casein hydrolysate for long periods. Albino Swiss mice were divided into eight groups. Diets contained 5% (undernourished) or 20% (controls) casein levels. For each group there were sub-groups ingesting whole or hydrolysed casein for 12 weeks. Infection with S. mansoni developed in half of the animals under each diet. All undernourished mice developed malabsorption. Low albuminemia was detected in infected animals independently of the protein level in the diet. However, albuminemia was lower in infected controls than in undernourished non-infected mice, suggesting a deficient liver protein synthesis. Infected mice fed on a 20% protein hydrolysed diet exhibited low weight gain and high mortality rates. On the other hand, non-infected mice ingesting the same diet had the highest body weights. We are investigating the hypothesis that infected mice, even when fed normal diets, are unable to metabolise large amounts of amino acids due to the liver lesions related to schistosomiasis and as a result die of hepatic coma. In some of them, the excessive accumulation of ammonia in the blood enhances the outcome of an encephalopathy.

Role of resident mast cells and macrophages in the neutrophil migration induced by LTB4, fMLP and C5a des arg.

In the present study, we have investigated the participation of resident peritoneal cells (macrophages and mast cells) in the neutrophil migration induced in rats by the intraperitoneal administration of LTB4, fMLP or C5a des arg. The intraperitoneal injection of LTB4 (10 nmol), fMLP (10 nmol) and C5a des arg (zymosan-activated plasma, 1 ml) caused an intense neutrophil migration compared to the saline control (1,000, 1,500 and 2,000%, respectively). An 83% depletion in the number of resident cells following peritoneal lavage reduced the LTB4-stimulated neutrophil migration by 73.6% without affecting that caused by fMLP and C5a des arg. Increasing the peritoneal macrophage population (236%) by pretreating the cavities with thioglycollate enhanced the neutrophil migration induced by LTB4 (129%), but did not alter that induced by fMLP and C5a des arg. Similarly, reducing the population of peritoneal mast cells containing toluidine-blue-staining granules by subchronically pretreating the cavities with compound 48/80 diminished the LTB4-induced NM by 69% but had no effect on the responses to fMLP and C5a des arg. Pretreating the animals with dexamethasone strongly inhibited (70%) the neutrophil migration induced by the intraperitoneal injection of LTB4, fMLP and C5a des arg. Indomethacin, BW A4C and NDGA had no such effect. The incubating medium from peritoneal macrophages and mast cells stimulated with LTB4 induced neutrophil migration when injected into the peritoneal cavity of rats. This migration was strongly reduced (70%) by treating the cells with dexamethasone. In contrast, stimulating the cells with fMLP or C5a des arg did not result in the release of any promigratory activity into the incubating fluid. Our results suggest that LTB4 induces neutrophil migration via a mechanism dependent on resident mast cells and macrophages while that induced by C5a des arg and fMLP seems to be independent of such cellular involvement. The neutrophil migration induced by LTB4 is apparently mediated by factor(s) whose release is blocked by dexamethasone. fMLP and C5a appear to cause in vivo migration by the formation of a concentration gradient and by a glucocorticoid-sensitive mechanism different from that stimulated by LTB4.

Intestinal protein absorption in malnourished mice with acute schistosomiasis mansoni.

Intestinal protein absorption was studied in undernourished albino Swiss mice with acute schistosomiasis mansoni. Undernutrition was induced by feeding mice with the Regional Basic Diet (RBD) ingested by human populations in Northeast Brazil, an experimental model previously developed in our laboratory. Weaning mice were infected with 40 cercariae and compared to undernourished non-infected mice and/or to infected mice fed a balanced control diet. Apparent and True Protein Absorption Coefficients were determined by nitrogen balance during five consecutive days ending at the 63rd day of the trial (acute phase of murine schistosomiasis). Fecal metabolic nitrogen (FMN) was determined after administration of a non-protein diet and was also calculated through linear regression. Our results showed a reduced protein absorption in non-infected RBD-fed mice as compared to mice fed a casein control diet. Infection with Schistosoma mansoni had apparently no effect on intestinal protein absorption in well-nourished mice. However, infection seemed to interfere with protein absorption in under-nourished animals, since the lowest absorption ratios have been detected among RBD-fed infected mice. A brief discussion is made on the advantages of using the method of linear regression for the determination of FMN.

Nutrition and acute schistosomiasis.

In northeast Brazil, nutritional deficiency diseases and schistosomiasis mansoni overlap. An experimental model, which reproduces the marasmatic clinical form of protein-energy malnutrition, was developed in this laboratory to study these interactions. Albino Swiss mice were fed with a food association ingested usually by human populations in northeast Brazil. This diet (Regional Basic Diet - RBD) has negative effects on the growth, food intake and protein utilization in infected mice (acute phase of murine schistosomiasis). Nitrogen balance studies have also shown that infection with Schistosoma mansoni has apparently no effect on protein intestinal absorption in well nourished mice. However, the lowest absorption ratios have been detected among RBD--fed infected animals, suggesting that superimposed schistosome infection aggravated the nutritional status of the undernourished host. The serum proteins electrophoretic pattern, as far as albumins are concerned, is quite similar for non-infected undernourished and infected well-fed animals. So, the significance of albumins as a biochemical indicator of the nutritional status of human populations residing in endemic foci of Manson's schistosomiasis, is discussable.