RESUMEN
Safe and reliable capture techniques for wild animals are important for ecologic studies and management operations. We assessed the efficiency of ketamine-medetomidine (K:M) injection and reversal with atipamezole. We anesthetized 67 raccoons (Procyon lotor; 34 males, 33 females) 103 times (individuals captured between one and five times) from April 2009-October 2010 in Mont-Orford Provincial Park, Quebec, Canada. We administered a 1:1 mixture by volume of ketamine and medetomidine by intramuscular injection. Mean (±SD) induction times for males and females were 6.1±2.8 and 6.6±3.7 min, respectively. Mean induction time was 2 min longer for juveniles than for adults (7.8±3.9 and 5.8±2.9 min, respectively) and longer in autumn than in spring for adults (7.7±3.8 and 5.4±2.9 min, respectively). Recovery time after administration of atipamezole was 9.6±3.8 and 8.4±4.4 min for males and females, respectively. Recovery time was longer in spring than in autumn (10.2±4 and 7.4±3.8 min, respectively) for adults. Induction time increased by 166% after five captures of the same individual. Immobilization did not affect body mass, adult survival, or female reproductive success. We suggest the K:M mixture used is a safe and reliable method for anesthetizing raccoons in field conditions.
Asunto(s)
Imidazoles/administración & dosificación , Inmovilización/veterinaria , Ketamina/administración & dosificación , Medetomidina/administración & dosificación , Mapaches/fisiología , Periodo de Recuperación de la Anestesia , Animales , Animales Salvajes/fisiología , Femenino , Inmovilización/métodos , Inyecciones Intramusculares/veterinaria , Ketamina/antagonistas & inhibidores , Masculino , Medetomidina/antagonistas & inhibidores , Estaciones del Año , Factores SexualesRESUMEN
Identifying natural barriers to movements of hosts associated with infectious diseases is essential for developing effective control strategies. Raccoon rabies variant (RRV) is a zoonosis of concern for humans because its main vector, the raccoon (Procyon lotor), is found near residential areas. In Québec, Canada, all cases of RRV found in raccoons since 2006 were detected on the eastern side of the Richelieu River, suggesting that this river acts as a barrier to gene flow and thus the potential for RRV to spread. The objectives of this study were to characterize the genetic structure of raccoon populations and assess the effect of the Richelieu River on the population structure in southern Québec, Canada. We also evaluated whether RRV spread potential differed between sex and at a larger spatial scale. Our analyses revealed a weak signal of genetic differentiation among individuals located on each side of the Richelieu River. At a larger spatial scale, genetic structuring was weak. Our results suggest that rivers might not always efficiently restrain raccoon movements and spread of RRV. We suggest that the difference in genetic structure found between sexes can be partly explained by male movements during the breeding season in winter, when ice bridges allow passage over most rivers in Québec.
RESUMEN
PURPOSE: To evaluate the feasibility and usefulness of a three-dimensional ultrasound (3D-US) image-guided system in identifying and tracking the tumor bed (TB) for planning and daily localization before radiation delivery for breast cancer. METHODS AND MATERIALS: Twenty breast cancer patients underwent two CT scans at the time of simulation and just before their boost. Three-dimensional ultrasound images were acquired immediately after the CT scans, to which the images were automatically fused. Three-dimensional ultrasound images were also acquired immediately before treatment. Spatial and temporal TB differences between CT and US were evaluated. RESULTS: The TB was not visible on US and CT in 1 subject who had and 1 subject who had not received chemotherapy before whole-breast radiotherapy. The mean (SD) TB volume overlap was 78% (14%). The mean centroid position of the TB on CT vs. US differed by 0.1, 0.2, and 0.4 mm in the anterior-posterior, left-right, and superior-inferior directions. The mean (SD) absolute radial displacement of the TB on each fraction from the treatment plan was 10.8 (6.3) mm. CONCLUSIONS: The TB was well visualized by US for the majority of patients. Clinically insignificant differences in the displacements calculated by paired CT vs. paired US demonstrate the feasibility of using 3D-US. The present study suggests that a 10-mm planning target volume margin could result in undercoverage of the clinical target volumes in 50% of treatments. Multimodality planning and image-guided radiotherapy with US potentially offers an accurate and non-ionizing solution for the daily definition of the TB position during partial-breast irradiation and boost treatments.
Asunto(s)
Neoplasias de la Mama/diagnóstico por imagen , Neoplasias de la Mama/radioterapia , Imagenología Tridimensional/métodos , Antineoplásicos/uso terapéutico , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Estudios de Factibilidad , Femenino , Humanos , Movimiento , Posicionamiento del Paciente , Tomografía Computarizada por Rayos X/métodos , Carga Tumoral , UltrasonografíaAsunto(s)
Cuidados en el Hogar de Adopción , Relaciones entre Hermanos , Hermanos/psicología , Niño , Custodia del Niño/organización & administración , Protección a la Infancia/legislación & jurisprudencia , Protección a la Infancia/psicología , Niños Huérfanos/legislación & jurisprudencia , Niños Huérfanos/psicología , Cuidados en el Hogar de Adopción/organización & administración , Cuidados en el Hogar de Adopción/psicología , Francia , Humanos , Psicología Infantil/legislación & jurisprudenciaRESUMEN
BACKGROUND & AIMS: Cystathionine beta-synthase (CBS) deficiency causes severe hyperhomocysteinemia, which confers diverse clinical manifestations, notably liver disease. To investigate this aspect of hyperhomocysteinemia, we performed a thorough investigation of liver pathology in CBS-deficient mice, a murine model of severe hyperhomocysteinemia. METHODS: The degree of liver injury and inflammation was assessed by histologic examination, by measurements of products of lipid peroxidation, and by formation of carbonyl groups on protein as a measure for the occurrence of protein oxidation. Analysis of profibrogenic, proinflammatory factors and cell apoptosis was performed by Western blots, real-time quantitative reverse-transcription polymerase chain reaction, caspase-3 activity, DNA laddering, and TUNEL assay. RESULTS: Histologic evaluation of liver specimens of 8- to 32-week-old CBS-deficient mice showed that CBS-deficient mice develop inflammation, fibrosis, and hepatic steatosis, concomitant with an enhanced expression of tissue inhibitor of metalloproteinase-1, alpha-smooth muscle actin, pro(alpha)1 collagen type I, transforming growth factor-beta1, and proinflammatory cytokines. Moreover, even if the proapoptotic protein Bax was dominantly expressed and Bcl-2 was down-regulated, caspase-3 was not activated, DNA laddering was not detected, and number of positive TUNEL cells was not increased in liver of CBS-deficient mice compared with wild-type mice. CONCLUSIONS: The results show that hyperhomocysteinemia in liver of CBS-deficient mice promotes oxidative stress, which may cause mitochondrial damage in association with activation of hepatic stellate cells, leading to liver injury. The absence of caspase-3 activation, DNA fragmentation, and TUNEL-positive cells shows that protective signals may counteract apoptotic signals in liver of CBS-deficient mice.
Asunto(s)
Cistationina betasintasa/genética , Hígado Graso/metabolismo , Homocistinuria/metabolismo , Cirrosis Hepática/metabolismo , Estrés Oxidativo/fisiología , Animales , Apoptosis/fisiología , Colágeno Tipo I/metabolismo , Modelos Animales de Enfermedad , Hígado Graso/patología , Hígado Graso/fisiopatología , Homocisteína/sangre , Homocistinuria/patología , Homocistinuria/fisiopatología , Hiperhomocisteinemia/metabolismo , Hiperhomocisteinemia/patología , Hiperhomocisteinemia/fisiopatología , Hígado/metabolismo , Hígado/patología , Cirrosis Hepática/patología , Cirrosis Hepática/fisiopatología , Ratones , Ratones Mutantes , ARN Mensajero/metabolismo , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Factor de Crecimiento Transformador beta1RESUMEN
Cystathionine beta synthase (CBS) is a crucial regulator of plasma concentrations of homocysteine. Severe hyperhomocysteinemia due to CBS deficiency confers diverse clinical manifestations, notably characteristic skeletal abnormalities. To investigate this aspect of hyperhomocysteinemia, we analyzed the skeleton of CBS-deficient mice, a murine model of severe hyperhomocysteinemia. Radiography, Alcian Blue/Alizarin Red S-stained whole skeletal preparations, and histological comparisons were used to determine the extent, pattern, and distribution of skeletal abnormalities in CBS-deficient mice. Disruption of the murine CBS gene leads to skeletal abnormalities, notably kyphoscoliosis, with temporal shortening of long bones due to impaired cartilage differentiation, albeit to differing degrees.
Asunto(s)
Huesos/anomalías , Cistationina betasintasa/deficiencia , Hiperhomocisteinemia/patología , Síndrome de Marfan/patología , Osteogénesis/fisiología , Escoliosis/patología , Animales , Huesos/diagnóstico por imagen , Huesos/enzimología , Cruzamiento/métodos , Cistationina betasintasa/genética , ADN/análisis , Modelos Animales de Enfermedad , Femenino , Genotipo , Homocisteína/sangre , Hiperhomocisteinemia/enzimología , Hiperhomocisteinemia/genética , Masculino , Síndrome de Marfan/enzimología , Síndrome de Marfan/genética , Ratones , Ratones Endogámicos , Ratones Noqueados , Radiografía , Escoliosis/enzimología , Escoliosis/genéticaRESUMEN
Cystathionine beta synthase (CBS) is a crucial regulator of plasma concentrations of homocysteine. Severe hyperhomocysteinemia due to CBS deficiency confers diverse clinical manifestations. Patients with severe hyperhomocysteinemia have fine hair and thin skin, but it is unclear whether these changes are related to CBS deficiency or are coincidental. To investigate these aspects of hyperhomocysteinemia, we characterized skin abnormalities of CBS-deficient mice, a murine model of severe hyperhomocysteinemia. Histological and histomorphometric analyses revealed that CBS-deficient mice have wrinkled skin with hyperkeratinosis of the epidermis and thinning of the dermis.
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Cistationina betasintasa/fisiología , Hiperhomocisteinemia/patología , Queratosis/enzimología , Queratosis/patología , Animales , Cistationina betasintasa/genética , Heterocigoto , Homocisteína/sangre , Homocigoto , Hiperhomocisteinemia/enzimología , Hiperhomocisteinemia/genética , Ratones , Ratones NoqueadosAsunto(s)
Arildialquilfosfatasa/genética , Hiperhomocisteinemia/enzimología , Hiperhomocisteinemia/genética , Hígado/enzimología , Animales , Secuencia de Bases , Cistationina betasintasa/genética , ADN/genética , Regulación hacia Abajo , Ratones , Ratones Mutantes , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Deficiency in cystathionine beta synthase (CBS) leads to high plasma homocysteine concentrations and causes hyperhomocysteinemia, a common risk factor for vascular disease, stroke and possibly neurodegenerative diseases. Various neuronal diseases have been associated with hyperhomocysteinemia, but the molecular mechanisms of homocysteine toxicity are unknown. We investigated the pathways involved in the pathological process, by analyzing differential gene expression in neuronal tissues. We used a combination of differential display and cDNA arrays to identify genes differentially expressed during hyperhomocysteinemia in brain of CBS-deficient mice. In this murine model of hyperhomocysteinemia, both plasma and brain homocysteine concentrations were high. Several genes were found to be differentially expressed in the brains of CBS-deficient mice, and the identities of some of these genes suggested that the SAPK/JNK pathway was altered in the brains of CBS-deficient mice. We therefore investigated the activation of proteins involved in the SAPK/JNK cascade. JNK and c-Jun were activated in the hippocampal neurones of CBS-deficient mice, suggesting that the SAPK/JNK pathway may play an important role in the development of neuronal defects associated with hyperhomocysteinemia.
Asunto(s)
Encéfalo/metabolismo , Hiperhomocisteinemia/metabolismo , MAP Quinasa Quinasa 4 , Neuronas/metabolismo , Transducción de Señal/fisiología , Factor de Transcripción Activador 2 , Animales , Encéfalo/citología , Química Encefálica , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Cistationina betasintasa/deficiencia , Cistationina betasintasa/genética , Modelos Animales de Enfermedad , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Hipocampo/citología , Hipocampo/metabolismo , Homocisteína/sangre , Homocisteína/metabolismo , Hiperhomocisteinemia/genética , Proteínas Quinasas JNK Activadas por Mitógenos , Ratones , Ratones Noqueados , Quinasas de Proteína Quinasa Activadas por Mitógenos/genética , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , Proteínas Quinasas Activadas por Mitógenos/genética , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Transducción de Señal/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Cystathionine beta-synthase (CBS) deficiency causes severe hyperhomocysteinemia and other signs of homocystinuria syndrome, in particular a premature atherosclerosis with multiple thrombosis. However, the molecular mechanisms by which homocysteine could interfere with normal cell function are poorly understood in a whole organ like the liver, which is central to the catabolism of homocysteine. We used a combination of differential display and cDNA arrays to analyze differential gene expression in association with elevated hepatic homocysteine levels in CBS-deficient mice, a murine model of hyperhomocysteinemia. Expression of several genes was found to be reproducibly abnormal in the livers of heterozygous and homozygous CBS-deficient mice. We report altered expression of genes encoding ribosomal protein S3a and methylthioadenosine phosphorylase, suggesting such cellular growth and proliferation perturbations may occur in homozygous CBS-deficient mice liver. Many up- or down-regulated genes encoded cytochromes P450, evidence of perturbations of the redox potential in heterozygous and homozygous CBS-deficient mice liver. The expression of various genes involved in severe oxidative processes was also abnormal in homozygous CBS-deficient mice liver. Among them, the expression of heme oxygenase 1 gene was increased, concomitant with overexpression of heme oxygenase 1 at the protein level. Commensurate with the difference in hepatic mRNA paraoxonase 1 abundance, the mean hepatic activity of paraoxonase 1, an enzyme that protects low density lipoprotein from oxidation, was 3-fold lower in homozygous CBS-deficient mice. Heterozygous CBS-deficient mice, when fed a hyperhomocysteinemic diet, have also reduced PON1 activity, which demonstrates the effect of hyperhomocysteinemia in the paraoxonase 1 activity.
Asunto(s)
Modelos Animales de Enfermedad , Expresión Génica , Hiperhomocisteinemia/genética , Hígado/metabolismo , Animales , Secuencia de Bases , Cartilla de ADN , Ratones , Ratones NoqueadosRESUMEN
Hyperhomocysteinemia, caused by a lack of cystathionine beta synthase (CBS), leads to elevated plasma concentrations of homocysteine. This is a common risk factor for atherosclerosis, stroke, and possibly neurodegenerative diseases. However, the mechanisms that link hyperhomocysteinemia due to CBS deficiency to these diseases are still unknown. Early biochemical studies describe developmental and adult patterns of transsulfuration and CBS expression in a variety of species. However, there is incomplete knowledge about the regional and cellular expression pattern of CBS, notably in the brain. To complete the previous data, we used in situ hybridization and Northern blotting to characterize the spatial and temporal patterns of Cbs gene expression during mouse development. In the early stages of development, the Cbs gene was expressed only in the liver and in the skeletal, cardiac, and nervous systems. The expression declined in the nervous system in the late embryonic stages, whereas it increased in the brain after birth, peaking during cerebellar development. In the adult brain, expression was strongest in the Purkinje cell layer and in the hippocampus. Immunohistochemical analyses showed that the CBS protein was localized in most areas of the brain but predominantly in the cell bodies and neuronal processes of Purkinje cells and Ammon's horn neurons.
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Encéfalo/metabolismo , Cistationina betasintasa/biosíntesis , Animales , Northern Blotting , Encéfalo/embriología , Encéfalo/crecimiento & desarrollo , Cistationina betasintasa/genética , Inmunohistoquímica , Hibridación in Situ , Ratones , Ratones Mutantes , ARN Mensajero/biosíntesisRESUMEN
UNLABELLED: Atrial fibrillation (AF) is accompanied by various changes in ion channels that cause atrial electrophysiological remodeling. The enzyme Na,K-ATPase is also a major cellular mechanism for the regulation of ion homeostasis. During AF, Na,K-ATPase may be regulated by synthesis of its alpha- and beta-subunits as well as changes in membrane fluidity. To test this hypothesis, we studied the effect of pacing-induced AF in sheep on atrial Na,K-ATPase alpha- and beta-subunits and on membrane fluidity as well. METHODS: A group of six sheep (AF group) was subjected to overdrive electrical stimulation of the right atrium in order to induce AF. A group of six sham operated sheep served as control. All paced sheep developed multiple episodes of sustained AF with a mean total duration of 110 min over a 2-hours period. Protein expression of Na,K-ATPase alpha- and beta-subunits in atrial microsomal membranes was assayed by Western blotting analysis. When significant changes in membrane expression were observed, transcriptional regulation was analysed by Northern blotting. Membrane fluidity was assessed on atrial microsomal fractions by anisotropy measurements using the fluorescent probe diphenylhexatriene. RESULTS: Atrial fibrillation enhanced the expression of the Na,K-ATPase beta1-subunit at both membrane and mRNA levels. Anisotropy values were higher in AF group than in control group, indicating a decreased fluidity of the membranes isolated from paced sheep atria. CONCLUSION: These data are the first evidence for an enhanced Na,K-ATPase beta1-subunit expression in membrane during AF. Membrane rigification represents a new factor of tachycardia-induced atrial remodeling.
Asunto(s)
Fibrilación Atrial/enzimología , Fluidez de la Membrana/fisiología , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Animales , Fibrilación Atrial/etiología , Northern Blotting , Western Blotting , Estimulación Cardíaca Artificial/efectos adversos , Cardioversión Eléctrica , Estimulación Eléctrica , Ácidos Grasos/metabolismo , Regulación de la Expresión Génica , Transporte Iónico/fisiología , Microsomas/enzimología , Subunidades de Proteína , Ovinos , ATPasa Intercambiadora de Sodio-Potasio/genética , Transcripción GenéticaRESUMEN
Disturbances of Na,K-ATPase activity are implicated in the pathophysiology of cerebral ischemia. Previous experiments have shown that EGb 761 protects NaK-ATPase activity against one hour of cerebral ischemia. In the brain however, the 3 isoenzymes responsible for Na,K-ATPase activity may be differentially affected by various times of ischemia. In the present study, we investigated the effect of a longer period of ischemia, and the protection provided by a pre-treatment with EGb 761 on each of the 3 cerebral NaK-ATPase isoenzymes. In control and EGb 761 pre-treated mice exposed to a 6 hr unilateral occlusion of the middle cerebral artery, Na,K-ATPase activity was decreased by 60% and lipid peroxidation was increased by 40% in the ipsilateral (ischemic) cortex compared to the contralateral one. In parallel, membrane integrity was altered. The alteration of NaK-ATPase activity, as a whole, resulted from a decrease in the activity of the 3 isoenzymes. The two isoenzymes of high ouabain affinity however, had their affinities decreased while the sensitivity of the lowest affinity isoenzyme was increased. Pre-treatment with EGb 761 abolished the differences observed between ipsi- and contralateral cortex, with the exception of the change in ouabain affinity of the low affinity isoenzyme. Ischemia also induced changes in Na,K-ATPase isoenzyme ouabain affinities in the contralateral cortex that where not prevented by EGb 761.