Unexpected appetitive events promote positive affective state in juvenile European sea bass.

Some animal species exhibit considerable physiological and behavioural alterations in response to captivity. It has been hypothesized, but rarely tested, that such changes reflect a negative affective state that is associated to this specific context. In the last years, judgement bias measures have emerged as reliable indicators of animal affective state, under the assumption that individuals in a negative affective state are more likely to evaluate ambiguous stimuli as negative and display therefore pessimistic behaviours. Here, we have developed a judgement bias task for juvenile European sea bass (Dicentrarchus labrax) aiming to measure optimism/pessimism in this marine species, which have previously been reported to show important dysregulations in captive settings. Our results show that juvenile sea bass exhibit a considerable bias towards pessimistic behaviours in laboratory settings. Furthermore, juveniles that received an unexpected positive event during the judgement bias test displayed more optimistic responses toward ambiguous stimuli as compared to control fish, indicating a positive change in their affective state induced by the appetitive experience. These results reveal a direct interaction of the internal affective state with decision-making processing under ambiguity in juvenile European sea bass, highlighting therefore the potential of judgement bias tests as a tool for the advancement and improvement of our understanding of welfare in finfish aquaculture.

Responsiveness of pituitary to galanin throughout the reproductive cycle of male European sea bass (Dicentrarchus labrax).

The neuropeptide galanin (Gal) is a putative factor regulating puberty onset and reproduction through its actions on the pituitary. The present study investigated the pituitary responsiveness to galanin and the patterns of galanin receptors (Galrs) expression throughout the reproductive cycle of two years old male European sea bass (Dicentrarchus labrax), an important aquaculture species. Quantitative analysis of pituitary and hypothalamus transcript expression of four galr subtypes revealed differential regulation according to the testicular developmental stage, with an overall decrease in expression from the immature stage to the mid-recrudescence stage. Incubation of pituitary cells with mammalian 1-29Gal peptide induced significant changes in cAMP concentration, with sensitivities that varied according to the testicular development stages. Furthermore 1-29Gal was able to stimulate both follicle stimulating hormone (Fsh) and luteinizing hormone (Lh) release from pituitary cell suspensions. The magnitude of the effects and effective concentrations varied according to reproductive stage, with generalized induction of Fsh and Lh release in animals sampled in January (full spermiation). The differential expression of galrs in pituitary and hypothalamus across the reproductive season, together with the differential effects of Gal on gonadotropins release in vitro strongly suggests the involvement of the galaninergic system in the regulation the hypothalamus-pituitary-gonad axis of male sea bass. This is to our knowledge the first clear evidence for the involvement of galanin in the regulation of reproduction in non-mammalian vertebrates.

[Satisfaction according to health care insurance systems in an emergency department]. / Satisfacción en un servicio de urgencias según el sistema de aseguramiento en salud.

INTRODUCTION: Health satisfaction is a fundamental measure of the quality of health services. This study aims to validate and analyse the results of a quality of care questionnaire to assess the level of satisfaction of patients attended in the emergency department of a high complexity hospital. METHODS: Observational, cross-sectional study, with a questionnaire designed to assess the quality of service and satisfaction at the end of care in the emergency department. Descriptive statistics of scale were established and presented, as well as determining the construct validity, overall reliability, internal and concurrent validity of an overall against a uni-dimensional scale. RESULTS: A total of 5,961 records were reviewed, most of them (77.3%) reported by patients in the Mandatory Health Plan. High levels of satisfaction overall and by subgroups were found. There were no significant differences between subgroups, with 86.8 for those with Pre-paid Medical Care Plan and 84.4 for mandatory health plan. Cronbach's alpha for the questionnaire was 0.90. CONCLUSIONS: The questionnaire proved to be reliable and valid in determining the quality and satisfaction with care. The results showed high levels of satisfaction overall and in the domains. A low consistency between the results of the multidimensional and unidimensional satisfaction scales suggests that there were aspects of satisfaction not investigated on the multidimensional scale. Ecologically-designed before and after studies are required to evaluate the effectiveness of interventions in satisfaction.

Actions of sex steroids on kisspeptin expression and other reproduction-related genes in the brain of the teleost fish European sea bass.

Kisspeptins are well known as mediators of the coordinated communication between the brain-pituitary axis and the gonads in many vertebrates. To test the hypothesis that gonadal steroids regulate kiss1 and kiss2 mRNA expression in European sea bass (a teleost fish), we examined the brains of gonad-intact (control) and castrated animals, as well as castrated males (GDX) and ovariectomized females (OVX) that received testosterone (T) and estradiol (E2) replacement, respectively, during recrudescence. In GDX males, low expression of kiss1 mRNA is observed by in situ hybridization in the caudal hypothalamus (CH) and the mediobasal hypothalamus (MBH), although hypothalamic changes in kiss1 mRNA levels were not statistically different among the groups, as revealed by real-time PCR. However, T strongly decreased kiss2 expression levels in the hypothalamus, which was documented in the MBH and the nucleus of the lateral recess (NRLd) in GDX T-treated sea bass males. Conversely, it appears that E2 evokes low kiss1 mRNA in the CH, while there were cells expressing kiss2 in the MBH and NRLd in these OVX females. These results demonstrate that kisspeptin neurons are presumably sensitive to the feedback actions of sex steroids in the sea bass, suggesting that the MBH represents a major site for sex steroid actions on kisspeptins in this species. Also, recent data provide evidence that both positive and negative actions occur in key factors involved in sea bass reproductive function, including changes in the expression of gnrh-1/gonadotropin, cyp19b, er and ar genes and sex steroid and gonadotropin plasma levels in this teleost fish.

Expression of kisspeptins and their receptors, gnrh-1/gnrhr-II-1a and gonadotropin genes in the brain of adult male and female European sea bass during different gonadal stages.

Kisspeptins play a critical role in the control of hypothalamic-gonadotropic function and puberty onset in mammals. Studies in fish have all supported the hypothesis that they might play similar roles in the reproduction of this animal group, however, their physiological relevance in the occurrence of key reproductive events still remains to be determined. This study examines the relative mRNA expression profiles of the duplicate kisspeptin system (kiss1, kiss2, gpr54-1b, and gpr54-2b) in the hypothalamus and pituitary of adult male and female sea bass (Dicentrarchus labrax L.) during different gonadal stages using qRT-PCR. We also report the changes in the expression levels of gnrh-1, gnrhr-II-1a, fshß, and lhß and the relationships observed between both kisspeptin and GnRH systems. Our data show clear sex differences in the dynamics of kisspeptin and kisspeptin receptor gene expression in the hypothalamus of sea bass during gonadal development. Overall, all four kisspeptin system genes increased either before or during the advanced stages of oogenesis and declined during atresia, exhibiting profiles that are identical to those observed for gnrhr-II-1a, fshß, lhß, and the gonadosomatic index (GSI). While the situation was not as clear in males, the high kiss2 expression levels observed in the hypothalamus during mid recrudescence suggest that it might be playing a role in the neuroendocrine signaling that regulates germ cell proliferation at the testicular level. In this sense, the proposed role attributed to kisspeptins as key factors in the onset of reproduction in fish receives an additional support from the data obtained in the present work. Nevertheless, further research is required to clarify their precise role in sea bass.

Validation of a lactic acid- and citric acid-based antimicrobial product for the reduction of Escherichia coli O157: H7 and Salmonella on beef tips and whole chicken carcasses.

The objectives of this study were to determine the effects of a lactic acid- and citric acid-based antimicrobial product on the reduction of Salmonella on whole broiler carcasses during processing and the reduction of Salmonella and Escherichia coli O157:H7 on beef trim. Freshly harvested broiler carcasses were inoculated with an inoculum of Salmonella strains to yield a 10(5) CFU/ml pathogen load on the surface of the carcass. The beef tips were inoculated as well with an inoculum of either E. coli O157:H7 or Salmonella to yield 10(4) CFU/100 cm(2). After 30 min for attachment, the broiler carcasses were treated with Chicxide applied for 5 s via a spray or immersed in Chicxide for 5, 10, or 20 s. Broiler carcasses were rinsed in poultry rinse bags with 400 ml of Butterfield's phosphate buffer in which Salmonella was enumerated from the diluents and Butterfield's phosphate. Chicxide significantly reduced Salmonella by 1.3 log CFU/ml with spray treatment and 2.3 log CFU/ml for all dip treatments. Following 30 min of attachment, the beef tips were placed into a spray cabinet with either Beefxide or sterilized water (control) and sprayed at 1 ft/2.5 s chain speed at 40 lb/in(2). The external surface of each beef tip was swabbed (100 cm(2)) to determine pathogen loads. Beefxide significantly reduced E. coli O157:H7 by 1.4 log CFU/100 cm(2) and Salmonella by 1.1 log CFU/100 cm(2) (P < 0.05) compared with the control samples.

Wall shear stress in intracranial self-expanding stents studied using ultra-high-resolution 3D reconstructions.

BACKGROUND AND PURPOSE: Imaging of intracranial stents is constrained by resolution limits of current clinical imaging techniques providing insufficient visualization of deployment details and impeding its use for computational hemodynamic (CHD) simulations. The purpose of our study was to evaluate whether ultra-high-resolution MicroCT scans can illuminate detailed aspects of realistic in vitro stent deployment and serve as a reliable basis for CHD simulations of blood flow through self-expanding intracranial stents. MATERIALS AND METHODS: A Neuroform Treo (NF) stent and an Enterprise (ENT) stent were deployed in identical straight polytetrafluoroethylene tubes filled with contrast agent. MicroCT scans were obtained at a spatial resolution of 14 mum and used for ultra-high-resolution 3D reconstructions. CHD simulations were performed, with particular emphasis on local flow behavior near the wall and struts. Flow differences between the geometrically different stents were studied. RESULTS: MicroCT data revealed strut prolapse near the markers for the closed-cell design (ENT) stent and at some of the unconnected vertices of the open-cell design (NF) stent, which also showed some misalignments. CHD simulations showed that reverse wall shear stress occurred near some of the strut vertices and markers for the NF but only near the markers for the ENT. CONCLUSIONS: This work demonstrates the feasibility of ultra-high-resolution MicroCT imaging in elucidating important details of intracranial stent deployment as a basis for accurate CHD simulations and in enabling a structural and hemodynamic study of realistically deployed stents with different geometry and design.

Gasto energético e ingesta de energía y macronutrientes en mujeres obesas en edad fértil / Energy expedinture and energy and macronutrient intake in obese women of fertile age

A 90 mujeres con sobrepeso y obesidad del área de atracción del Programa de Atención Integral en Salud, PAIS, se les midió la tasa metabólica basal por calorimetría indirecta y se les evaluó tanto el nivel de actividad física como la ingesta de energía y macronutrientes. Se observó una correlación positiva entre el Indice de Masas Corporal y la tasa metabólica basal de estas mujeres. Aunque se encontró diferencia significativa entre el gasto metabólico basal estimado por la ecuación de Harris-Benedict y el gasto metabólico basal medido por calorimetría indirecta, no se encontró diferencia significativa en el total de energía expedida obtenida por ambos métodos. En estas pacientes con sobrepeso no se encontró diferencia significativa entre el gasto energético total y la ingesta energética diaria obtenida, lo que sugiere un estado de equilibrio energético adaptado a la nueva condición de fisiológica. Según la distribución de macronutrientes fue la grasa la que excedió la recomendación dietética diaria.

A group of 90 women with overweight and obesity, residing in the area covered by the Program for Integral Attention in Health (PAIS) participated in this study. The following measurements were made: basal metabolic rate by indirect calorimetry, level of physical activity and intake of energy and macronutrients. There was a positive correlation between Body Mass Index and the basal metabolic rate for these women. Although a significant difference was found between the average basal metabolic expenditure of the group, as estimated by the Harris-Benedict equation and average basal metabolic expenditure as measured by indirect calorimetry, there was no significant difference between total energy expenditure as estimated by both methods. No significant difference was found between average energy expenditure and average energy intake for these women, suggesting a state of energy balance which is an adaption to the new physiological condition. According to the distribution of macronutrients, fat intake exceeded nutritional recommendations.

Presence of viral proteins in drinkable water--sufficient condition to consider water a vector of viral transmission?

In order to determine the role of water as a possible vector for transmission of the most prevalent enteric viruses affecting infantile populations, 226 water samples were collected from Facatativa's (Colombian municipality located 30km away from Bogotá) water works in the years 2000, 2002, and 2005. The samples were clarified and virus was concentrated by filtering and ultrafiltering techniques. The presence of viral protein (VP) was assessed by enzyme immunoassay method (EIA) and viral RNA presence was detected by reverse trascriptase and polymerase chain reaction (RT-PCR). Using these techniques, one sample positive for Astrovirus (HAstV) was found in a sample collected from the river that supplies the aqueduct, two samples positive for Norovirus (NV) from fresh treated potable water and 13 samples positive for Rotavirus (RV), some in water from the plant during treatment and others from treated fresh water. RT-PCR inhibitors were also found in water samples obtained from the plant and in the fresh treated water. No inhibitors were found in the river water. VP, but no nucleic acid, was detected in the water samples at different stages of treatment, thus suggesting that the virus might have been complete and infectious at some stage prior to water purification.

[Prevalence of sub-clinical vitamin A deficiency and malnutrition in slum children in Maraicabo - Venezuela]. / Prevalencia de deficiencia subclínica de vitamina A y desnutrición en niños marginales de Maracaibo--Venezuela.

The present cross sectional study was carried out to estimate the prevalence of vitamin A deficiency among children by means of clinics and conjunctival impression cytology (CIC), and nutritional status by anthropometric indicators H//A, W//A, W//H. The study population included 157 children 2-6 y old, from urban and rural slums of Maracaibo, Venezuela, Conjunctival impression cytology was performed by ICEPO standard procedure. Z-score was applied to anthropometric data with reference values of NCHS-WHO. No evidence of clinical or ophthalmologic signs of vitamin A deficiency were detected. The prevalence of subclinical vitamin A deficiency, as detected by abnormal CIC, was 35.4%, being higher in rural children (48.3%). These prevalence values are higher than the criteria laid down by WHO/UNICEF to indicate a public health problem (> 20%). Mild or moderate protein-energy global malnutrition and stunting were detected in 36.1% and 44.6% of children, respectively. Abnormal CIC was indistinctly observed (approximately equal to 35%) as much in children with adequate nutrition as in malnourished ones. There was no significant difference in the distribution of the CIC results in relation to nutritional status. The findings indicate that CIC and Z-score of nutritional anthropometric data are useful to characterize the risk of vitamin A deficiency and of malnutrition in communities. Beside the implementation of an integral nutritional program which includes supplementation, food fortification and dietary diversification, improvement of socio-economic and sanitation conditions and also the educational level, with emphasis on nutritional and health education, are highly recommended.

Field emission scanning electron microscopy and freeze-fracture transmission electron microscopy of mouse cerebellar synaptic contacts.

Samples of albino mice were processed by the cryofracture method for scanning electron microscopy and examined with the field emission scanning electron microscope (FESEM). Freeze-etching direct replicas of mice cerebellar cortex were also studied with the transmission electron microscope (FFTEM), as a complementary technique for obtaining higher resolution, three-dimensional correlative images of cerebellar synaptic contacts. At the granular, Purkinje cells and molecular layers, the cryofracture method for FESEM selectively removed the neuroglial cell investment, facilitating the visualization of the outer and inner surfaces of cerebellar synaptic contacts. In addition, FFTEM showed the real extension of perisynaptic neuroglial investment. The outer surface of mossy fiber rosettes and their digitiform processes were seen at the granular layer, making flat and invaginated synaptic contacts with the granule cell dendrites. At the molecular layer, the longitudinal traject of parallel fibers or nonsynaptic segments and their synaptic varicosities were characterized. These latter established synaptic contacts with Purkinje dendritic spines. Fractured parallel fiber endings showed the SE-I images of clustered spheroidal synaptic vesicles and mitochondria and the surrounding cotton-like appearance of Bergmann glial cell cytoplasm. Climbing fibers showed a characteristic crossing-over bifurcation pattern in the white matter and in the three-layer structure of cerebellar cortex, formation of tendril collaterals in the granular layer, topographical relationship with Purkinje cell soma and retrograde collaterals in the molecular layer. The climbing fiber synaptic relationship with Purkinje dendritic spines was characterized, by means of FFTEM, by the presence of large synaptic endings and aggregation of intramembrane particles at the P and E faces of presynaptic endings, characteristic of excitatory synapses.

Further observations on cerebellar climbing fibers. A study by means of light microscopy, confocal laser scanning microscopy and scanning and transmission electron microscopy.

The intracortical pathways of climbing fibers were traced in several vertebrate cerebella using light microscopy, confocal laser scanning microscopy, scanning and transmission electron microscopy. They were identified as fine fibers up to 1(micron thick, with a characteristic crossing-over bifurcation pattern. Climbing fiber collaterals were tridimensionally visualized forming thin climbing fiber glomeruli in the granular layer. Confocal laser scanning microscopy revealed three types of collateral processes at the interface between granular and Purkinje cell layers. Scanning electron microscopy showed climbing fiber retrograde collaterals in the molecular layer. Asymmetric synaptic contacts of climbing fibers with Purkinje dendritic spines and stellate neuron dendrites were characterized by transmission electron microscopy. Correlative microscopy allowed us to obtain the basic three-dimensional morphological features of climbing fibers in several vertebrates and to show with more accuracy a higher degree of lateral collateralization of these fibers within the cerebellar cortex. The correlative microscopy approach provides new views in the cerebellar cortex information processing.

Further observations on cerebellar climbing fibers. A study by means of light microscopy, confocal laser scanning microscopy and scanning and transmission electron microscopy.

The intracortical pathways of climbing fibers were traced in several vertebrate cerebella using light microscopy, confocal laser scanning microscopy, scanning and transmission electron microscopy. They were identified as fine fibers up to 1(micron thick, with a characteristic crossing-over bifurcation pattern. Climbing fiber collaterals were tridimensionally visualized forming thin climbing fiber glomeruli in the granular layer. Confocal laser scanning microscopy revealed three types of collateral processes at the interface between granular and Purkinje cell layers. Scanning electron microscopy showed climbing fiber retrograde collaterals in the molecular layer. Asymmetric synaptic contacts of climbing fibers with Purkinje dendritic spines and stellate neuron dendrites were characterized by transmission electron microscopy. Correlative microscopy allowed us to obtain the basic three-dimensional morphological features of climbing fibers in several vertebrates and to show with more accuracy a higher degree of lateral collateralization of these fibers within the cerebellar cortex. The correlative microscopy approach provides new views in the cerebellar cortex information processing.

Immunolocalization of inhibin in the mammary gland of rats treated with hCG.

Human chorionic gonadotropin (hCG) induces profuse lobular development in the mammary gland of young virgin rats. To clarify whether the effect of hCG is locally mediated by inhibin, a non-steroidal glycoprotein, we detected its localization immunocytochemically in the mammary gland with polyclonal antibodies against the alpha- and beta-chains. Virgin female Sprague-Dawley rats were sacrificed after treatment with a daily IP injection of 100 IU hCG for 5, 10, 15, or 21 days of treatment or 20 days after the last injection. Whereas the mammary gland of control animals did not contain immunoreactive inhibin, hCG treatment induced the expression of inhibin in the cytoplasm of alveolar cells but not in ductal cells. The reaction became evident by Day 10 of treatment and reached its maximal intensity by Day 15. Thereafter, the reaction became evident in the stroma, which exhibited maximal positivity by Day 20. Once hCG treatment was terminated, the mammary gland regressed to its pre-treatment condition, appearing similar both in morphology and inhibin content to that of control animals. The expression of this glycoprotein hormone in the mammary gland after hCG administration at the time of maximal lobulolveolar development, and its diffusion towards the stroma during regression, suggest a critical role of inhibin as a modulator of mammary growth and differentiation.

Transient accumulations of glycosaminoglycans in rat and chicken cerebellar cortex during development.

Modifications of glycosaminoglycans at neuropile of rat and chicken cerebellum during development were histochemically studied. The application of Alcian Blue staining techniques and enzymatic degradations permitted to reveal in both species that in earlier stages of cerebellar development hyaluronic acid is present throughout neuropile of entire cerebellum but it accumulated preferentially at the medullary region and around precursory Purkinje cells where it showed a mucoid-like appearance. This substance was related with cell migration and aligning processes. At the middle of cerebellar development, around 2nd postnatal week in rat and 12-16 embryonary days in chick, a new polyanionic transient accumulation, presumably chondroitinsulphate, became present at the medullary region following the longitudinal axis of folium and limiting the forming granular layer, being this substance mainly related with polarity processes by controlling or guiding the growing cones of afferent fibers, which enter massively to cerebellar cortex. It disappeared as myelination progressed. Also from the middle stage of development onward, beside glycosaminoglycans, other polyanionic substances were present at the molecular and granular layer neuropile and at the cytoplasm of some nerve cells. These macromolecules were rather related with nerve cell differentiation and maturation.

Histochemical distribution of acid glycosaminoglycans in rat cerebellum during postnatal development: neuropile.

Developmentally regulated modifications of glycosaminoglycans (GAGs) in the central nervous system (CNS) have suggested that also in the CNS, these compounds might participate in morphogenesis and nerve cell differentiation. However very few studies have been reported concerning the regional distribution of these compounds by histochemical techniques. We have used the Alcian Blue staining method also in conjunction with enzymatic digestion and with a technique which allowed the measurement of the degree of GAG sulphatation. The combination of the three techniques showed that during the first week GAGs, presumed to be hyaluronic acid, are localized throughout the neuropile of the entire cerebellum and especially in the medullary region. Sulphated glycans appear later in the medullary region (particularly at the border between the medullary region and the internal granular layer) and in all the layers of cerebellum (in particular around the Purkinje and deep cerebellar nuclei neurons and possibly in the cerebellar glomeruli). Sulphated glycans in the medullary region disappear around the 12th day when myelination starts. The transient presence of glycoproteins in the molecular layer was also detected.

Histochemical demonstration of cytoplasmic glycosaminoglycans in the macroneurons of the human central nervous system.

The presence of glycosaminoglycans (GAG) has been histochemically demonstrated in the CNS of various mammalian species. They have been related with some nerve functions as neurotransmitters storage and synaptic transmission. In the present paper, the histochemical properties of nerve cell cytoplasmic GAG were studied in several regions of adult human CNS. Samples of brain cortex, pons, upper medulla, and cerebellar cortex obtained by autopsy from subjects not dying after neurological diseases were fixed by immersion in glutaraldehyde, dehydrated with ethanol, and embedded in paraffin. The sections were stained with Alcian blue solutions adjusted to pH 2.5, 4.0, and 5.7. To the latter solution MgCl2 was added in increasing concentration from 0.05 to 1.2 M. Testicular hyaluronidase, neuraminidase, and ribonuclease were applied on simultaneous sections with their respective controls. The sequence of these reactions allowed us to demonstrate the presence of hyaluronic acid along chondroitin-4- and/or 6-sulphate in the cytoplasm of most nerve cells. The sulphated GAG showed certain variability in the various regions studied related specially with their grade of sulphation.