RESUMEN
In collaboration with the American College of Veterinary Radiology.
Asunto(s)
Radiología , Animales , Humanos , Radiografía , Estados UnidosRESUMEN
Yessotoxin (YTX) is a marine phycotoxin produced by dinoflagellates and accumulated in filter feeding shellfish. YTX content in shellfish is regulated by many food safety authorities to protect human health, although currently no human intoxication episodes have been unequivocally related to YTX presence in food. The immune system has been proposed as one of the target organs of YTX due to alterations of lymphoid tissues and cellular and humoral components. The aim of the present study was to explore subacute immunotoxicity of YTX in rats by evaluating the haematological response, inflammatory cytokine biomarkers and the presence of YTX-induced structural alterations in the spleen and thymus. The results showed that repeated administrations of YTX caused a decrease of lymphocyte percentage and an increase of neutrophil counts, a reduction in interleukine-6 (IL-6) plasmatic levels and histopathological splenic alterations in rats after four intraperitoneal injections of YTX at doses of 50 or 70 µg/kg that were administered every 4 days along a period of 15 days. Therefore, for the first time, subacute YTX-immunotoxicity is reported in rats, suggesting that repeated exposures to low amounts of YTX might also suppose a threat to human health, especially in immuno-compromised populations.
Asunto(s)
Inmunotoxinas/toxicidad , Oxocinas/toxicidad , Mariscos/análisis , Animales , Biomarcadores/sangre , Dinoflagelados/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Contaminación de Alimentos , Inocuidad de los Alimentos , Interleucina-6/sangre , Recuento de Linfocitos , Linfocitos/citología , Linfocitos/efectos de los fármacos , Venenos de Moluscos , Neutrófilos/citología , Oxocinas/inmunología , Ratas , Ratas Sprague-Dawley , Bazo/efectos de los fármacos , Bazo/patología , Timo/efectos de los fármacos , Timo/patología , Factor de Necrosis Tumoral alfa/sangreRESUMEN
Yessotoxins (YTX) and azaspiracids (AZAs) are marine toxins produced by phytoplanktonic dinoflagellates that get accumulated in filter feeding shellfish and finally reach human consumers through the food web. Both toxin classes are worldwide distributed, and food safety authorities have regulated their content in shellfish in many countries. Recently, YTXs and AZAs have been described as compounds with subacute cardiotoxic potential in rats owed to alterations of the cardiovascular function and ultrastructural heart damage. These molecules are also well known in vitro inducers of cell death. The aim of this study was to explore the presence of cardiomyocyte death after repeated subacute exposure of rats to AZA-1 and YTX for 15 days. Because autophagy and apoptosis are often found in dying cardiomyocytes, several autophagic and apoptotic markers were determined by western blot in heart tissues of these rats. The results showed that hearts from YTX-treated rats presented increased levels of the autophagic markers microtubule-associated protein light chain 3-II (LC3-II) and beclin-1, nevertheless AZA-1-treated hearts evidenced increased levels of the apoptosis markers cleaved caspase-3 and -8, cleaved PARP and Fas ligand. Therefore, while YTX-induced damage to the heart triggers autophagic processes, apoptosis activation occurs in the case of AZA-1. For the first time, activation of cell death signals in cardiomyocytes is demonstrated for these toxins with in vivo experiments, which may be related to alterations of the cardiovascular function.
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Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Toxinas Marinas/toxicidad , Miocitos Cardíacos/efectos de los fármacos , Oxocinas/toxicidad , Compuestos de Espiro/toxicidad , Animales , Biomarcadores/metabolismo , Western Blotting , Femenino , Toxinas Marinas/administración & dosificación , Venenos de Moluscos , Oxocinas/administración & dosificación , Ratas , Ratas Sprague-Dawley , Compuestos de Espiro/administración & dosificación , Factores de Tiempo , Pruebas de Toxicidad Subaguda/métodosRESUMEN
Yessotoxin (YTX) is a marine phycotoxin produced by dinoflagellates and accumulated in filter feeding shellfish. Although no human intoxication episodes have been reported, YTX content in shellfish is regulated by many food safety authorities due to their worldwide distribution. YTXs have been related to ultrastructural heart damage in vivo, but the functional consequences in the long term have not been evaluated. In this study, we explored the accumulative cardiotoxic potential of YTX in vitro and in vivo. Preliminary in vitro evaluation of cardiotoxicity was based on the effect on hERG (human ether-a-go-go related gene) channel trafficking. In vivo experiments were performed in rats that received repeated administrations of YTX followed by recordings of electrocardiograms, arterial blood pressure, plasmatic cardiac biomarkers, and analysis of myocardium structure and ultrastructure. Our results showed that an exposure to 100 nM YTX for 12 or 24 h caused an increase of extracellular surface hERG channels. Furthermore, remarkable bradycardia and hypotension, structural heart alterations, and increased plasma levels of tissue inhibitor of metalloproteinases-1 were observed in rats after four intraperitoneal injections of YTX at doses of 50 or 70 µg/kg that were administered every 4 days along a period of 15 days. Therefore, and for the first time, YTX-induced subacute cardiotoxicity is supported by evidence of cardiovascular function alterations related to its repeated administration. Considering international criteria for marine toxin risk estimation and that the regulatory limit for YTX has been recently raised in many countries, YTX cardiotoxicity might pose a health risk to humans and especially to people with previous cardiovascular risk.
Asunto(s)
Cardiotoxinas/toxicidad , Enfermedades Cardiovasculares/metabolismo , Corazón/efectos de los fármacos , Oxocinas/toxicidad , Animales , Células CHO , Cardiotoxicidad , Cardiotoxinas/administración & dosificación , Cardiotoxinas/química , Células Cultivadas , Cricetulus , Canal de Potasio ERG1/metabolismo , Humanos , Inyecciones Intraperitoneales , Conformación Molecular , Venenos de Moluscos , Oxocinas/administración & dosificación , Oxocinas/química , Ratas , Ratas Sprague-DawleyRESUMEN
Azaspiracids (AZAs) are marine toxins produced by Azadinium spinosum that get accumulated in filter feeding shellfish through the food-web. The first intoxication was described in The Netherlands in 1990, and since then several episodes have been reported worldwide. Azaspiracid-1, AZA-2, and AZA-3 presence in shellfish is regulated by food safety authorities of several countries to protect human health. Azaspiracids have been related to widespread organ damage, tumorogenic properties and acute heart rhythm alterations in vivo but the mechanism of action remains unknown. Azaspiracid toxicity kinetics in vivo and in vitro suggests accumulative effects. We studied subacute cardiotoxicity in vivo after repeated exposure to AZA-1 by evaluation of the ECG, arterial blood pressure, plasmatic heart damage biomarkers, and myocardium structure and ultrastructure. Our results showed that four administrations of AZA-1 along 15 days caused functional signs of heart failure and structural heart alterations in rats at doses ranging from 1 to 55 µg/kg. Azaspiracid-1 altered arterial blood pressure, tissue inhibitors of metalloproteinase-1 plasma levels, heart collagen deposition, and ultrastructure of the myocardium. Overall, these data indicate that repeated exposure to low amounts of AZA-1 causes cardiotoxicity, at doses that do not induce signs of other organic system toxicity. Remarkably, human exposure to AZAs considering current regulatory limits of these toxins may be dangerously close to clearly cardiotoxic doses in rats. These findings should be considered when human risk is estimated particularly in high cardiovascular risk subpopulations.
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Insuficiencia Cardíaca/inducido químicamente , Toxinas Marinas/toxicidad , Compuestos de Espiro/toxicidad , Animales , Presión Arterial/efectos de los fármacos , Biomarcadores/sangre , Cardiotoxicidad , Colágeno/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Insuficiencia Cardíaca/sangre , Insuficiencia Cardíaca/fisiopatología , Frecuencia Cardíaca/efectos de los fármacos , Mediadores de Inflamación/sangre , Miocardio/metabolismo , Miocardio/ultraestructura , Ratas Sprague-Dawley , Medición de Riesgo , Factores de Tiempo , Pruebas de Toxicidad SubagudaRESUMEN
Phycotoxins are marine toxins produced by phytoplankton that can get accumulated in filter feeding shellfish. Human intoxication episodes occur due to contaminated seafood consumption. Okadaic acid (OA) and dynophysistoxins (DTXs) are phycotoxins responsible for a severe gastrointestinal syndrome called diarrheic shellfish poisoning (DSP). Yessotoxins (YTXs) are marine toxins initially included in the DSP class but currently classified as a separated group. Food safety authorities from several countries have regulated the content of DSPs and YTXs in shellfish to protect human health. In mice, OA and YTX have been associated with ultrastructural heart damage in vivo. Therefore, this study explored the potential of OA, DTX-1 and YTX to cause acute heart toxicity. Cardiotoxicity was evaluated in vitro by measuring hERG (human èter-a-go-go gene) channel activity and in vivo using electrocardiogram (ECG) recordings and cardiac damage biomarkers. The results demonstrated that these toxins do not exert acute effects on hERG channel activity. Additionally, in vivo experiments showed that these compounds do not alter cardiac biomarkers and ECG in rats acutely. Despite the ultrastructural damage to the heart reported for these toxins, no acute alterations of heart function have been detected in vivo, suggesting a functional compensation in the short term.
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Cardiotoxicidad , Ácido Ocadaico/toxicidad , Oxocinas/toxicidad , Piranos/toxicidad , Animales , Células CHO , Cardiotoxicidad/sangre , Cardiotoxicidad/fisiopatología , Cricetinae , Cricetulus , Canal de Potasio ERG1 , Electrocardiografía , Canales de Potasio Éter-A-Go-Go/fisiología , Femenino , Venenos de Moluscos , Péptido Natriurético Encefálico/sangre , Ratas , Ratas Sprague-Dawley , Troponina I/sangre , Troponina T/sangreRESUMEN
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.
RESUMEN
Azaspiracids (AZAs) are marine biotoxins produced by the dinoflagellate Azadinium spinosum that accumulate in several shellfish species. Azaspiracid poisoning episodes have been described in humans due to ingestion of AZA-contaminated seafood. Therefore, the contents of AZA-1, AZA-2 and AZA-3, the best-known analogs of the group, in shellfish destined to human consumption have been regulated by food safety authorities of many countries to protect human health. In vivo and in vitro toxicological studies have described effects of AZAs at different cellular levels and on several organs, however, AZA target remains unknown. Very recently, AZAs have been demonstrated to block the hERG cardiac potassium channel. In this study, we explored the potential cardiotoxicity of AZA-2 in vivo. The effects of AZA-2 on rat electrocardiogram (ECG) and cardiac biomarkers were evaluated for cardiotoxicity signs besides corroborating the hERG-blocking activity of AZA-2. Our results demonstrated that AZA-2 does not induce QT interval prolongation on rat ECGs in vivo, in spite of being an in vitro blocker of the hERG cardiac potassium channel. However, AZA-2 alters the heart electrical activity causing prolongation of PR intervals and the appearance of arrhythmias. More studies will be needed to clarify the mechanism by which AZA-2 causes these ECG alterations; however, the potential cardiotoxicity of AZAs demonstrated in this in vivo study should be taken into consideration when evaluating the possible threat that these toxins pose to human health, mainly for individuals with pre-existing cardiovascular disease when regulated toxin limits are exceeded.
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Arritmias Cardíacas/inducido químicamente , Furanos/toxicidad , Piranos/toxicidad , Animales , Biomarcadores/sangre , Células CHO/efectos de los fármacos , Cricetulus , Canal de Potasio ERG1 , Electrocardiografía , Canales de Potasio Éter-A-Go-Go/genética , Canales de Potasio Éter-A-Go-Go/metabolismo , Femenino , Miocardio/metabolismo , Técnicas de Placa-Clamp , Bloqueadores de los Canales de Potasio/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
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 â¼20% 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.
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Envejecimiento , Neuronas Colinérgicas/metabolismo , Trastornos del Conocimiento/patología , Prosencéfalo/patología , Receptor de Factor de Crecimiento Nervioso/metabolismo , Animales , Modelos Animales de Enfermedad , PerrosRESUMEN
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.
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Envejecimiento/sangre , Enfermedad de Alzheimer/veterinaria , Péptidos beta-Amiloides/sangre , Fragmentos de Péptidos/sangre , Envejecimiento/psicología , Enfermedad de Alzheimer/sangre , Enfermedad de Alzheimer/psicología , Animales , Biomarcadores/sangre , Estudios de Casos y Controles , Cognición , Modelos Animales de Enfermedad , Perros , Femenino , MasculinoRESUMEN
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.
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Enfermedad de Alzheimer/patología , Trastornos del Conocimiento/patología , Locus Coeruleus/patología , Degeneración Nerviosa/patología , Neuronas/patología , Norepinefrina/metabolismo , Factores de Edad , Envejecimiento/metabolismo , Envejecimiento/patología , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/fisiopatología , Animales , Biomarcadores/análisis , Biomarcadores/metabolismo , Canidae/anatomía & histología , Canidae/metabolismo , Recuento de Células , Trastornos del Conocimiento/metabolismo , Trastornos del Conocimiento/fisiopatología , Modelos Animales de Enfermedad , Perros , Femenino , Inmunohistoquímica , Locus Coeruleus/metabolismo , Locus Coeruleus/fisiopatología , Masculino , Degeneración Nerviosa/metabolismo , Degeneración Nerviosa/fisiopatología , Neuronas/metabolismo , Placa Amiloide/metabolismo , Placa Amiloide/patología , Corteza Prefrontal/metabolismo , Corteza Prefrontal/patología , Corteza Prefrontal/fisiopatología , Especificidad de la Especie , Tirosina 3-Monooxigenasa/análisis , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
A 7-year-old spayed female English Cocker Spaniel was examined because of a 1-week history of lethargy, stumbling over objects and circling, and the presence of two tonic-clonic generalised seizures two days before presentation. The neurological signs suggested a lesion involving the right forebrain. Computed tomography revealed the presence of two intracranial masses, one located inside the right lateral ventricle and the other located in the right frontal lobe attached to the falx cerebri. Because of the poor prognosis, the owner refused to continue with the therapy and the dog was euthanised. On postmortem examination one mass was diagnosed histologically as a meningioma and the other as a papilloma of the choroid plexus. Information in the veterinary literature on multiple malignancies affecting the central nervous system is very limited. To the best of the authors' knowledge, the association of meningioma and choroid plexus papilloma has never been reported either in the human or in the veterinary medical literature.
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Enfermedades de los Perros/patología , Meningioma/veterinaria , Papiloma del Plexo Coroideo/veterinaria , Animales , Encéfalo/patología , Perros , Femenino , Meningioma/diagnóstico , Meningioma/patología , Papiloma del Plexo Coroideo/diagnóstico , Papiloma del Plexo Coroideo/patologíaRESUMEN
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.
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Envejecimiento , Enfermedad de Alzheimer/veterinaria , Péptidos beta-Amiloides/metabolismo , Enfermedades de los Perros/patología , Degeneración Nerviosa/veterinaria , Neuronas/patología , Núcleos del Rafe/patología , Serotonina/metabolismo , Enfermedad de Alzheimer/patología , Análisis de Varianza , Animales , Recuento de Células , Perros , Inmunohistoquímica , Neuronas/fisiología , Núcleos del Rafe/metabolismo , Triptófano Hidroxilasa/metabolismoRESUMEN
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.