Measuring haemolysis in cattle serum by direct UV-VIS and RGB digital image-based methods.

A simple, rapid procedure is required for the routine detection and quantification of haemolysis, one of the main sources of unreliable results in serum analysis. In this study, we compared two different approaches for the rapid determination of haemolysis in cattle serum. The first consisted of estimating haemolysis via a simple direct ultraviolet-visible (UV-VIS) spectrophotometric measurement of serum samples. The second involved analysis of red, green, blue (RGB) colour data extracted from digital images of serum samples and relating the haemoglobin (Hb) content by means of both univariate (R, G, B and intensity separately) and multivariate calibrations (R, G, B and intensity jointly) using partial least squares regression and artificial neural networks. The direct UV-VIS analysis and RGB-multivariate analysis using neural network methods were both appropriate for evaluating haemolysis in serum cattle samples. The procedures displayed good accuracy (mean recoveries of 100.7 and 102.1%, respectively), adequate precision (with coefficients of variation from 0.21 to 2.68%), limit of detection (0.14 and 0.21 g L-1, respectively), and linearity of up to 10 g L-1.

Higher rates of first episode psychosis in immigrants admitted in inpatient unit at southwest Spain.

We present data on the incidence of admissions for first episode psychosis in a region of southern Spain. All consecutive cases of admissions to the psychiatric hospitalization unit   due to psychosis were selected. The incidence rates for first episode psychosis among immigrants and non-immigrants between two years were calculated.  Incidence rate ratio of first episode of psychosis was higher in immigrants (IRR 5.95 95% CI 3.8-9.3 p<0.001) and also in individuals from Sub-Saharan Africa (IRR: 30.09 95% CI:16.2-55.8 p<0.001). The results reflect the risk that immigrants have a greater risk of being hospitalized than non-immigrants.

Influence of Haemolysis on the Mineral Profile of Cattle Serum.

Haemolysis of serum samples is the leading cause of preanalytical errors in clinical laboratories. Little is known about the potential alterations in the concentrations of mineral elements in haemolyzed serum and the phenomenon has not been specifically studied in bovine serum samples. We investigate how haemolysis affects the mineral content of bovine samples. We used ICP-MS to measure the concentrations of 12 mineral elements (Ca, Co, Cr, Cu, Fe, Mg, Mn, Mo, Ni, P, Se and Zn) in bovine whole blood, serum and gradually haemolyzed samples and observed significant differences between the different types of samples, particularly in the Fe and Zn concentrations. However, in practice, the high interindividual variability makes it difficult to establish whether a given value corresponds to normal or haemolyzed samples. In response to this problem, we propose to consider that a result is significantly biased when the haemolysis threshold (the degree of haemolysis above which the concentration of an element in serum is significantly altered) of a given element is surpassed. The haemolysis threshold values for the different elements considered were found as follows: 0.015 g Hb L-1 for Fe, 2 g for Zn, 4 g for Cr and 8 g for Ca, Se and Mo.

Neurotensin in reward processes.

Neurotensin (NTS) is a neuropeptide neurotransmitter expressed in the central and peripheral nervous systems. Many studies over the years have revealed a number of roles for this neuropeptide in body temperature regulation, feeding, analgesia, ethanol sensitivity, psychosis, substance use, and pain. This review provides a general survey of the role of neurotensin with a focus on modalities that we believe to be particularly relevant to the study of reward. We focus on NTS signaling in the ventral tegmental area, nucleus accumbens, lateral hypothalamus, bed nucleus of the stria terminalis, and central amygdala. Studies on the role of NTS outside of the ventral tegmental area are still in their relative infancy, yet they reveal a complex role for neurotensinergic signaling in reward-related behaviors that merits further study. This article is part of the special issue on 'Neuropeptides'.

Manipulations of Central Amygdala Neurotensin Neurons Alter the Consumption of Ethanol and Sweet Fluids in Mice.

The central nucleus of the amygdala plays a significant role in alcohol use and other affective disorders; however, the genetically-defined neuronal subtypes and projections that govern these behaviors are not well known. Here we show that neurotensin neurons in the central nucleus of the amygdala of male mice are activated by in vivo ethanol consumption and that genetic ablation of these neurons decreases ethanol consumption and preference in non-ethanol-dependent animals. This ablation did not impact preference for sucrose, saccharin, or quinine. We found that the most robust projection of the central amygdala neurotensin neurons was to the parabrachial nucleus, a brain region known to be important in feeding behaviors, conditioned taste aversion, and alarm. Optogenetic stimulation of projections from these neurons to the parabrachial nucleus is reinforcing, and increases ethanol drinking as well as consumption of sucrose and saccharin solutions. These data suggest that this central amygdala to parabrachial nucleus projection influences the expression of reward-related phenotypes and is a novel circuit promoting consumption of ethanol and palatable fluids.SIGNIFICANCE STATEMENT Alcohol use disorder (AUD) is a major health burden worldwide. Although ethanol consumption is required for the development of AUD, much remains unknown regarding the underlying neural circuits that govern initial ethanol intake. Here we show that ablation of a population of neurotensin-expressing neurons in the central amygdala decreases intake of and preference for ethanol in non-dependent animals, whereas the projection of these neurons to the parabrachial nucleus promotes consumption of ethanol as well as other palatable fluids.

A fatal case of Babesia divergens infection in Northwestern Spain.

We describe a fatal case caused by the intra-erythrocytic Babesia divergens parasite in an elderly woman. This is the third case of fatal babesiosis reported in the last 15 years in Europe, and the only one in a patient with an intact spleen.

Integration of Plasticity Mechanisms within a Single Sensory Neuron of C. elegans Actuates a Memory.

Neural plasticity, the ability of neurons to change their properties in response to experiences, underpins the nervous system's capacity to form memories and actuate behaviors. How different plasticity mechanisms act together in vivo and at a cellular level to transform sensory information into behavior is not well understood. We show that in Caenorhabditis elegans two plasticity mechanisms-sensory adaptation and presynaptic plasticity-act within a single cell to encode thermosensory information and actuate a temperature preference memory. Sensory adaptation adjusts the temperature range of the sensory neuron (called AFD) to optimize detection of temperature fluctuations associated with migration. Presynaptic plasticity in AFD is regulated by the conserved kinase nPKCε and transforms thermosensory information into a behavioral preference. Bypassing AFD presynaptic plasticity predictably changes learned behavioral preferences without affecting sensory responses. Our findings indicate that two distinct neuroplasticity mechanisms function together through a single-cell logic system to enact thermotactic behavior. VIDEO ABSTRACT.

Miocarditis asociada a infección por Campylobacter jejuni / Myocarditis associated with Campylobacter jejuni infection

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The effect of water salinity on wood breakdown in semiarid Mediterranean streams.

Saline streams occur naturally and they are distributed worldwide, particularly in arid and semiarid regions, but human activities have also increased their number in many parts of the world. Little attention has been paid to assess increasing salt effects on organic matter decomposition. The objectives of this study were to analyse wood breakdown rates and how salinity affects them in 14 streams that exemplify a natural salinity gradient. We also analysed the effect of this gradient on changes in wood chemical composition, fungal biomass and microbial activity. Our results showed low breakdown rates (0.0010-0.0032 d(-1)), but they fell within the same range as those reported in freshwater streams when a similar woody substrate was used. However, salinity had a negative effect on the breakdown rates and fungal biomass along the salinity gradient, and led to noticeable changes in wood composition. Water salinity did not affect microbial activity estimated using hydrolysis of fluorescein diacetate. Variation in breakdown rates and fungal biomass across streams was mediated mainly by salinity, and later by stream discharge. Despite the role of fungi in stick breakdown, the potential wood abrasion by salts must be analysed in detail to accurately understand the effect of increasing salinity on organic matter breakdown. Finally, our results indicate that increased salinity worldwide by human activities or by the global warming would imply organic matter breakdown and mineralisation slowing down, even in natural saline streams. However, because many variables are implicated, the final effect of climatic change on organic matter decomposition in streams is difficult to predict.

Neprilysin is poorly expressed in the prefrontal cortex of aged dogs with cognitive dysfunction syndrome.

Neprilysin (NEP) is the principal amyloid ß (A ß ) degrading peptidase; this activity may protect against Alzheimer's disease (AD), the most important age-related neurodegenerative process. The aim of this work was to analyze NEP mRNA expression in the frontal cortex of dogs with and without canine cognitive dysfunction syndrome (CDS), which is considered a natural model for AD. Expression of canine cerebral NEP mRNA was assessed by RT-PCR followed by qPCR in young, aged-cognitively unimpaired (CU), and aged-cognitively impaired (CI) dogs. On average, aged-CI dogs showed 80% (P < 0.01) lower expression levels of NEP mRNA than their aged-CU counterparts. Furthermore, the standard deviation of the qPCR measurements was more than 6 times higher in the cognitively healthy animals (young and aged-CU) than in the aged-CI group. Another interesting find is the determination of a positive correlation between NEP expression and the number of cholinergic neurons in basal telencephalon, indicating a probable connection between both events in these types of neurodegeneration processes. These results suggest that high expression levels of NEP might be a protective factor for canine CDS and, most likely, for other A ß -associated neurodegenerative diseases, such as AD.

Establishing physico-chemical reference conditions in Mediterranean streams according to the European Water Framework Directive.

Type-specific physico-chemical reference conditions are required for the assessment of ecological status in the Water Framework Directive context, similarly to the biological and hydro-morphological elements. This directive emphasises that natural variability of quality elements in high status (reference condition) needs to be quantified. Mediterranean streams often present a marked seasonal pattern in hydrological, biological and geochemical processes which could affect physico-chemical reference conditions. This study establishes general physico-chemical reference conditions (oxygenation, nutrient, salinity and acidification conditions) for different Mediterranean stream types. 116 potential reference sites located in 23 Mediterranean catchments in Spain were sampled in spring, summer and autumn in 2003. All sites were subjected to a screening method for the selection of reference sites in Mediterranean streams (Mediterranean Reference Criteria) and classified using a pre-established stream typology that establishes five different stream types (temporary streams, evaporite-calcareous at medium altitude, siliceous headwaters, calcareous headwaters and large watercourses). Reference conditions (reference value and reference threshold equivalents to high-good class boundary) were calculated using two different methods according to the availability of reference sites: the reference site 75th percentile approach of all reference sites and the 25th percentile of the population approach. The majority of the studied potential reference sites (76 out of 116) were selected as reference sites. Regarding type-specific reference conditions, only siliceous headwaters could be considered different from the rest of stream types because lower conductivity and pH. All reference stream types presented seasonal differences as regards some parameters, except for temporary streams due to the high natural variation of this stream type. For those parameters which presented seasonal differences in a specific stream type, the least restrictive values were proposed as reference conditions.

Expression of p75(NTR), a marker for basal forebrain cholinergic neurons, in young and aged dogs with or without cognitive dysfunction syndrome.

The canine cognitive dysfunction syndrome (CDS) has been identified as a natural model for Alzheimer's disease (AD). We have used unbiased stereology to estimate the total number of basal forebrain cholinergic neurons expressing the nerve growth factor p75(NTR) receptor in young, aged cognitively-unimpaired (CU) and aged cognitively-impaired (CI) dogs. Aged-CI dogs showed a ∼20% decrement (p = 0.009) in p75(NTR) neurons compared to both the young and the aged-CU animals. These results suggest that the basal forebrain cholinergic system is affected in dogs with CDS and provide additional support for the use this canine syndrome as a model for AD research.

Plasma ß-amyloid peptides in canine aging and cognitive dysfunction as a model of Alzheimer's disease.

Aging dogs naturally demonstrate cognitive impairment and neuropathology that model early Alzheimer's disease (AD). In particular, there is evidence that canine cognitive dysfunction syndrome (CDS) in aged dogs is accompanied by cortical deposition of Aß peptides and neurodegeneration. Plasma Aß levels have been examined in humans as putative biomarkers for AD, but to date, no similar studies have been conducted for canine dementia. The aim of the present study was to assess plasma Aß1-42 and Aß1-40 levels in a blind study using pet dogs that were either successfully aging or exhibiting CDS. The severity of cognitive impairment was assessed using an owner-based questionnaire. On average, young dogs presented significantly higher plasma levels of Aß1-42 and Aß1-40 than aged, cognitively unimpaired dogs. Notably, among aged dogs, the levels of Aß1-42 and the Aß42/40 ratio were significantly higher in those showing mild cognitive impairment than in either cognitively unimpaired or severely affected dogs. These results suggest that increased plasma Aß1-42 levels and Aß42/40 ratio could be a biomarker for canine cognitive dysfunction, which is considered an excellent natural model of early AD.

Dogs with canine counterpart of Alzheimer's disease lose noradrenergic neurons.

Degeneration of noradrenergic neurons in the locus ceruleus is a well-described feature of Alzheimer's disease (AD). In spite of extensive utilization of the dog as a model for human degenerative diseases, there is no data on the response to aging of the noradrenergic system in dogs. We have used modern unbiased stereology to estimate the total number of A6-A7 noradrenergic neurons in normal, aged dogs and dogs with the canine counterpart of AD. In small-breed dogs with no cognitive impairments, the total mean number of tyrosine hydroxylase immunolabeled A6-A7 neurons was 17,228+/-1655, with no differences between young and aged dogs. In contrast, aged dogs with cognitive impairments exhibited a significant reduction in the total number of A6-A7 neurons (13,487+/-1374; P=0.001). Additionally, we found a negative correlation between the number of A6-A7 neurons and the extent of beta-amyloid deposits in the prefrontal cortex. These results suggest that the canine model could be useful in exploring the potential benefits of noradrenergic drugs for the treatment of AD.

Visceral leishmaniasis with cardiac involvement in a dog: a case report.

A dog presented with cutaneous nodules, enlarged lymph nodes and oedema in limbs, face and abdomen. The diagnosis of visceral leishmaniasis was established by identification of Leishmania amastigotes within macrophages from skin and popliteal lymph node biopsies. At necropsy, lesions were found in different organs, but it was particularly striking to observe large areas of pallor in the myocardium. Histological examination revealed an intense chronic inflammatory reaction in many organs, and numerous macrophages were found to contain amastigote forms of Leishmania. The inflammatory reaction was especially severe in the heart, where large areas of the myocardium appeared infiltrated with huge numbers of mononuclear immune cells, causing cardiac muscle atrophy and degeneration. Despite the severe inflammation, the number of parasitized macrophages was low in the myocardium, as revealed by immunohistochemical staining of Leishmania amastigotes. Because cardiac involvement is not usually described in this condition, this dog represents a very rare case of canine visceral leishmaniasis with affection of the myocardium.

Beta-amyloid cortical deposits are accompanied by the loss of serotonergic neurons in the dog.

Dogs may naturally suffer an age-related cognitive impairment that has aroused a great deal of interest, even beyond the field of the veterinary clinic. This canine senile dementia reproduces several key aspects of Alzheimer's disease (AD), including the presence of beta-amyloid (A beta) deposits in the cerebral cortex, neurodegeneration, and learning and memory impairments. In the present study, we have used unbiased stereological procedures to estimate the number of the dorsal and median raphe nuclei (DRN and MRN, respectively) serotonergic neurons immunolabeled with an anti-tryptophan hydroxylase (TrH) monoclonal antibody in young and aged dogs without A beta cortical deposits and in aged dogs with A beta cortical deposits. The estimated total number of TrH-labeled neurons (mean +/- SD) was 94,790 +/- 26,341 for the DRN and 40,404 +/- 8,692 for the MRN. The statistical analyses revealed that aged dogs with A beta cortical pathology had 33% fewer serotonergic neurons in the DRN and MRN than aged dogs without A beta cortical deposits (108,043 +/- 18,800 vs. 162,242 +/- 39,942, respectively; P = 0.01). In contrast, no significant variations were found between young and aged dogs without A beta cortical deposits. These results suggest that degeneration of the serotonergic neurons could be involved in the cognitive damage that accompanies A beta cortical pathology in the dog and reinforce the use of the canine model for exploring the potential mechanisms linking the cortical A beta pathology and serotonergic neurodegeneration that occurs during the course of AD.

The immunohistochemical localization of neuronal nitric oxide synthase in the basal forebrain of the dog.

The present work describes for the first time the anatomical distribution of neuronal nitric oxide synthase (nNOS) immunoreactivity and NADPH-d activity in the basal forebrain of the dog. As in other species, small, intensely nNOS-immunoreactive cells were seen within the olfactory tubercle, caudate nucleus, putamen, nucleus accumbens and amygdala. In addition, a population of mixed large and small nNOS positive cells was found in the medial septum, diagonal band and nucleus basalis overlapping the distribution of the magnocellular cholinergic system of the basal forebrain. Our results show that the distribution of NOS containing neurons in these nuclei in the dog is more extensive and uniform than that reported in rodents and primates. When double labeling of nNOS and NADPH-d was performed in the same tissue section most neurons were double labeled. However, a considerable number of large perikarya in the diagonal band and nucleus basalis appeared to be single labeled for nNOS. Thought a certain degree of interference between the two procedures could not be completely excluded, these findings suggest that NADPH-d histochemistry, which is frequently used to show the presence of NOS, underestimates the potential of basal forebrains neurons to produce nitric oxide. In addition, a few neurons mainly localized among the fibers of the internal capsule, appeared to be labeled only for NADPH-d. These neurons could be expressing a different isoform of NOS, not recognized by our anti-nNOS antibody, as has been reported in healthy humans and AD patients.

Aplicación de la encuesta dietética por inventario de almacén a la vigilancia alimentaria en instituciones / Dietary survey based on warehouse foodstuffs inventory applied to nutritional surveillance in institutions

Se propuso la aplicación de encuestas dietéticas por inventario de alimentos en almacén como complemento al método vigente. Se realizó la investigación durante el primer semestre de 1997 en 11 centros de la provincia de Cienfuegos. Se aplicaron encuestas semanales para la evaluación continua de la oferta de nutrientes. Este procedimiento es sencillo y barato para el monitoreo de la dieta y permite mostrar gráficamente los cambios en el tiempo. Se concluye que este tipo de encuesta puede ser aplicado en la alimentación institucional