Efecto fisiológico y genotóxico de la deficiencia de cobre inducida por Molibdeno en bovinos

Molybdenosis is a disease caused by the depressing effect of molybdenumn (Mo) on the physiological availability of Cu. The present study was carried out in order to analyze the ability of Mo to cause damage on the DNA integrity and changes in membrane fatty acids by oxidative damage. Holstein male calves were fed a Mo-supplemented diet for 9 months. Variables evaluated were plasma Cu concentration, erythrocyte Cu content and SOD activity, comet assay and analysis of the fatty acid composition of erythrocyte membranes. The statistical design was a completely randomized with one single factor and two replications. Copper plasma concentration, erythrocyte copper concentration and Cu/Zn SOD activity were analyzed using the t test. Chi-square test was used to compare the number of cells with DNA damage and one-way analysis of variance and Tukey test (p£0.05) for fatty acid composition and lipid peroxidation. Results showed that Mo in the diet induced a depletion of hepatic Cu storage, a decrease of Cu plasma and erythrocyte levels, a fall in Cu/Zn-SOD activity, changes in membrane fatty acids composition and DNA damage. These results are in agreement with the three phases model of copper deficiency and validate the occurrence of molybdenosis or secondary hypocuprosis. Further studies will be necessary to explore the mechanisms involved in the DNA damage and to distinguish primary molybdenum toxicosis from the molybdenum-induced copper deficiency.La molibdenosis es una enfermedad causada por el efecto depresivo del molibdeno (Mo) en la disponibilidad fisioló- gica del Cu. El presente estudio se llevó a cabo para analizar la capacidad del Mo para causar daño en la integridad del ADN y cambios en los ácidos grasos de la membrana a causa de daño oxidativo. Se alimentaron becerros machos Holstein con una dieta suplementada con Mo durante 9 meses. Las variables evaluadas fueron la concentración de Cu en el plasma, el contenido de Cu en eritrocitos y la actividad SOD, un ensayo cometa y el análisis de la composición de ácidos grasos de las membranas de eritrocitos. El diseño estadístico fue completamente al azar con un solo factor y dos réplicas. Se analizaron la concentración de cobre en el plasma, la concentración de cobre en eritrocitos y la actividad Cu/Zn-SOD con la prueba de t. Se utilizó la prueba de Chi cuadrado para comparar el número de células con daño del ADN y el análisis de varianza en un sentido y la prueba de Tukey (p£0.05) para la composición de ácidos grasos y la peroxidación de lípidos. Los resultados mostraron que el Mo en la dieta indujo una reducción en el almacenaje hepático de Cu, una disminución de los niveles de Cu en el plasma y eritrocitos, una caída en la actividad Cu/Zn-SOD, cambios en la composición de ácidos grasos de la membrana y daño del ADN. Estos resultados coinciden con el modelo de tres fases de deficiencia del cobre y validan la ocurrencia de molibdenosis o hipocuprosis secundaria. Serán necesarios más estudios para explorar los mecanismos involucrados en el daño del ADN y distinguir la toxicosis primaria por molibdeno a partir de la deficiencia de cobre inducida por el mismo.Fil: Picco, Sebastian Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de la Plata. Facultad de Cs.veterinarias. Departamento de Cs.basicas. Catedra de Fisiologia; ArgentinaFil: Ponzzinibio, M. Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Mattioli, Guillermo Alberto. Universidad Nacional de la Plata. Facultad de Cs.veterinarias. Departamento de Cs.basicas. Catedra de Fisiologia; ArgentinaFil: Rosa, Diana. Universidad Nacional de la Plata. Facultad de Cs.veterinarias. Departamento de Cs.basicas. Catedra de Fisiologia; ArgentinaFil: Minatel, Leonardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Fazzio, Luis Emilio. Universidad Nacional de la Plata. Facultad de Cs.veterinarias. Departamento de Cs.basicas. Catedra de Fisiologia; ArgentinaFil: Seoane, Analia Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentin

Characterization of low weight gain in calves with hypocuprosis

Una de las consecuencias productivas más graves de la hipocuprosis bovina es la menor ganancia diaria de peso (< GDP) en terneros. Su diagnóstico se realiza empleando valores de riesgo de indicadores indirectos. Estos indicadores presentan una gran variabilidad que posiblemente se deba a que están absorbiendo la variabilidad individual en el grupo. El objetivo de este trabajo fue caracterizar a los animales que están perdiendo peso dentro del grupo con baja performance, con el fin de establecer valores de riesgo más ajustados. Se realizaron cinco ensayos de suplementación hasta la aparición de < GDP. Todos los animales con < GDP (n: 150) se analizaron en conjunto por rangos de GDP. Se diferenciaron sólo dos subgrupos con GDP mayores y menores de 400 gr. Esto sugiere que en los grupos afectados se combinan terneros con alta y baja performance. Estos últimos representan apenas el 16 % del total y presentan concentraciones de Cu en plasma, sangre entera y glóbulos rojos menores de 15, 30 y 60 μg/dl respectivamente, y un porcentaje de hematocrito menor del 35 %. Los valores hallados ayudan a mejorar el valor diagnóstico de los indicadores de riesgo y ayudan a explicar su variabilidad.The low average daily gain (&lt; ADG) is one of the most serious consequences of hypocuprosis in calves. Diagnosis are made using risk values of indirect markers. These markers present a wide variations, which probably occurs because they absorb the individual variations within the group. The aim of this study was characterize the animals with &lt; ADG within the low performance group, in order to find more accurate risk values. Five supplementation trials were done until &lt; ADG occur. All animals with &lt; ADG (n: 150) were analyzed for ADG ranges. Two subgroups with ADG higher and lower than 400 gr were differentiated. It suggest that the affected group combined calves with high and low performance. The last represents just 16 % of total animals, and they had plasma, whole blood and red cell copper concentrations lower than 15, 30 and 60 μg/dl respectively, and PCV lower than 35 %. These results improve the diagnostic values of risk markers and also they help to explain their variability.Facultad de Ciencias Veterinaria

Effects of copper parenteral supplementation during the last gestational period of cows in a Cu deficient area

En una zona con incidencia endémica de hipocuprosis se evaluó el efecto terapéutico de la suplementación parenteral con Cu en vacas con carencia moderada y severa durante su último tercio de gestación. El grupo suplementado (n = 20) recibió 125 mg de edetato de Cu, mientras que el grupo control permaneció sin suplementación (n = 20). Se evaluaron las cupremias de las madres antes de la suplementación, al parto y cuando los terneros promediaban los tres meses de edad. Los terneros provenientes de ambos grupos se identificaron y pesaron al nacimiento y se sangraron a los tres meses para evaluar su estatus de Cu. Cuando se suplementaron madres con carencia moderada (ensayo A), las cupremias resultaron normales al parto pero descendieron a valores de deficiencia severa a los tres meses (< 30 ug/dl). En ese momento los terneros de madres suplementadas presentaron mayores concentraciones de Cu en plasma y glóbulos rojos, pero igualmente eran indicativas de carencia severa debido al agotamiento de sus reservas hepáticas. Cuando se suplementaron madres con carencia severa (ensayo B) los terneros ya no se diferenciaban por su estatus de Cu a los tres meses de edad. En ambos ensayos los terneros nacieron y crecieron hasta los tres meses sin diferenciarse por su peso, pero sus estatus de Cu indicaron la existencia de un riesgo clínico y productivo importante en una zona donde la primera suplementación se realiza al destete, con 6 a 7 meses de edad. Se concluye que la suplementación de las madres con 125 mg de Cu en el último tercio de la gestación puede resultar insuficiente en la protección de los terneros, especialmente si éstos no son suplementados durante su período de cría.In an endemically Cu deficient area, the effects of parenteral Cu supplementation during last third of the gestation were evaluated in cows with moderate and severe Cu deficiency. Supplemented group (n = 20) was injected with 125 mg of Cu edetate, another group (n = 20) remained without supplementation as control. Copper plasma level was measured in cows before supplementation, at calving and when calves were three months old. Calves were identified and weighted at calving, and blood samples were taken at three months old to evaluate their Cu status. When moderate deficient cows were supplemented (assay A), they had normal Cu plasma levels at calving, but returned to severe deficiency (Cu–plasma < 30 ug/dl) when calves were three months old. At that moment, calves from supplemented mothers had higher concentrations of Cu in plasma and red cells, although they were indicative of severe deficiency due to liver storage depletion. When severe deficient cows were injected (assay B), the Cu status of calves could not be differentiated between groups. In both assays, calves were born and grown until three months old without showing differences in body weight, although their Cu status indicated important clinical and productive risks in an area where the first Cu supplementation occurs at weaning, when calves are 6–7 months old. It is concluded that 125 mg Cu supplementation in cows during the last third of the gestation may be insufficient in calves’ protection, in particular if calves do not receive Cu supplementation during the breeding period.Facultad de Ciencias Veterinaria

Valor diagnóstico dos parâmetros de cobre para prever o crescimento de bezerros lactantes em pastagem nativas na Argentina

A study was conducted to evaluate the predictive diagnostic value of different copper (Cu) parameters as indicators of average daily gain (ADG) in growing calves. The effects in calves of cow Cu supplementation in the last one-third gestation period were also evaluated. Five supplementation trials, with a total of 300 calves, were carried out. Two groups of 30 calves were randomly assigned to each trial, one group was parenterally supplemented (SG) and the other was not supplemented (NSG). Trials began when calves were three-month-old and ended at weaning time. At each sampling calves were weighed and blood was taken to determine Cu concentrations in plasma, Whole Blood (WB), Red Cells (RC) and Packed Cell Volume (PCV). Liver samples from six animals of each group were taken both at the beginning and at the end of the trial. In two trials the mothers of the SG received Cu supplementation at the last one- third gestation period. Four of the five trials exhibited low ADG in the NSGs. In these groups, plasma Cu concentration decreased rapidly before low ADG was detected, which occurred with values remaining below 25μg/dl. The decrease of RC Cu concentration was considerably slow. WB showed an intermediate position. PCV in the SGs was higher than in the NSGs in all trials. Cow supplementation was insufficient to generate a liver storage able to last after calves reached the 3 months of age. These data could be useful to predict the risk of low ADG in grazing calves.Facultad de Ciencias Veterinaria

Tomotherapy after Pleurectomy/Decortication or Biopsy for Malignant Pleural Mesothelioma Allows the Delivery of High Dose of Radiation in Patients with Intact Lung

n/

Sweden

Molybdenosis is a disease caused by the depressing effect of molybdenumn (Mo) on the physiological availability of Cu. The present study was carried out in order to analyze the ability of Mo to cause damage on the DNA integrity and changes in membrane fatty acids by oxidative damage. Holstein male calves were fed a Mo-supplemented diet for 9 months. Variables evaluated were plasma Cu concentration, erythrocyte Cu content and SOD activity, comet assay and analysis of the fatty acid composition of erythrocyte membranes. The statistical design was a completely randomized with one single factor and two replications. Copper plasma concentration, erythrocyte copper concentration and Cu/Zn SOD activity were analyzed using the t test. Chi-square test was used to compare the number of cells with DNA damage and one-way analysis of variance and Tukey test (p≤0.05) for fatty acid composition and lipid peroxidation. Results showed that Mo in the diet induced a depletion of hepatic Cu storage, a decrease of Cu plasma and erythrocyte levels, a fall in Cu/Zn-SOD activity, changes in membrane fatty acids composition and DNA damage. These results are in agreement with the three phases model of copper deficiency and validate the occurrence of molybdenosis or secondary hypocuprosis. Further studies will be necessary to explore the mechanisms involved in the DNA damage and to distinguish primary molybdenum toxicosis from the molybdenum-induced copper deficiency.La molibdenosis es una enfermedad causada por el efecto depresivo del molibdeno (Mo) en la disponibilidad fisioló- gica del Cu. El presente estudio se llevó a cabo para analizar la capacidad del Mo para causar daño en la integridad del ADN y cambios en los ácidos grasos de la membrana a causa de daño oxidativo. Se alimentaron becerros machos Holstein con una dieta suplementada con Mo durante 9 meses. Las variables evaluadas fueron la concentración de Cu en el plasma, el contenido de Cu en eritrocitos y la actividad SOD, un ensayo cometa y el análisis de la composición de ácidos grasos de las membranas de eritrocitos. El diseño estadístico fue completamente al azar con un solo factor y dos réplicas. Se analizaron la concentración de cobre en el plasma, la concentración de cobre en eritrocitos y la actividad Cu/Zn-SOD con la prueba de t. Se utilizó la prueba de Chi cuadrado para comparar el número de células con daño del ADN y el análisis de varianza en un sentido y la prueba de Tukey (p≤0.05) para la composición de ácidos grasos y la peroxidación de lípidos. Los resultados mostraron que el Mo en la dieta indujo una reducción en el almacenaje hepático de Cu, una disminución de los niveles de Cu en el plasma y eritrocitos, una caída en la actividad Cu/Zn-SOD, cambios en la composición de ácidos grasos de la membrana y daño del ADN. Estos resultados coinciden con el modelo de tres fases de deficiencia del cobre y validan la ocurrencia de molibdenosis o hipocuprosis secundaria. Serán necesarios más estudios para explorar los mecanismos involucrados en el daño del ADN y distinguir la toxicosis primaria por molibdeno a partir de la deficiencia de cobre inducida por el mismo.Instituto de Genética VeterinariaFacultad de Ciencias Veterinaria

Radical Hemithoracic Radiotherapy Induces Systemic Metabolomics Changes That Are Associated with the Clinical Outcome of Malignant Pleural Mesothelioma Patients

Simple SummaryRadical hemithoracic radiotherapy represents a promising new advance in the field of radiation oncology and encouraging results have been achieved in the treatment of malignant pleural mesothelioma patients. This study showed that this radiotherapy modality produces significant changes in serum metabolomics profile mainly affecting arginine and polyamine biosynthesis pathways. Interestingly, individual metabolomics alterations were found associated with the clinical overall survival outcome of the radiotherapy treatment. These results highlight metabolomics profile analysis as a powerful prognostic tool useful to better understand the mechanisms underlying the interpatients variability and to identify patients who may receive the best benefit from this specific radiotherapy treatment.Radical hemithoracic radiotherapy (RHRT) represents an advanced therapeutic option able to improve overall survival of malignant pleural mesothelioma patients. This study aims to investigate the systemic effects of this radiotherapy modality on the serum metabolome and their potential implications in determining the individual clinical outcome. Nineteen patients undergoing RHRT at the dose of 50 Gy in 25 fractions were enrolled. Serum targeted metabolomics profiles were investigated at baseline and the end of radiotherapy by liquid chromatography and tandem mass spectrometry. Univariate and multivariate OPLS-DA analyses were applied to study the serum metabolomics changes induced by RHRT while PLS regression analysis to evaluate the association between such changes and overall survival. RHRT was found to affect almost all investigated metabolites classes, in particular, the amino acids citrulline and taurine, the C14, C18:1 and C18:2 acyl-carnitines as well as the unsaturated long chain phosphatidylcholines PC ae 42:5, PC ae 44:5 and PC ae 44:6 were significantly decreased. The enrichment analysis showed arginine metabolism and the polyamine biosynthesis as the most perturbed pathways. Moreover, specific metabolic changes encompassing the amino acids and acyl-carnitines resulted in association with the clinical outcome accounting for about 60% of the interpatients overall survival variability. This study highlighted that RHRT can induce profound systemic metabolic effects some of which may have a significant prognostic value. The integration of metabolomics in the clinical assessment of the malignant pleural mesothelioma could be useful to better identify the patients who can achieve the best benefit from the RHRT treatment

Biological Pathways Associated With the Development of Pulmonary Toxicities in Mesothelioma Patients Treated With Radical Hemithoracic Radiation Therapy: A Preliminary Study.

Radical hemithoracic radiotherapy (RHR), after lung-sparing surgery, has recently become a concrete therapeutic option for malignant pleural mesothelioma (MPM), an asbestos-related, highly aggressive tumor with increasing incidence and poor prognosis. Although the toxicity associated to this treatment has been reduced, it is still not negligible and must be considered when treating patients. Genetic factors appear to play a role determining radiotherapy toxicity. The aim of this study is the identification of biological pathways, retrieved through whole exome sequencing (WES), possibly associated to the development of lung adverse effects in MPM patients treated with RHR. The study included individuals with MPM, treated with lung-sparing surgery and chemotherapy, followed by RHR with curative intent, and followed up prospectively for development of pulmonary toxicity. Due to the strong impact of grade 3 pulmonary toxicities on the quality of life, compared with less serious adverse events, for genetic analyses, patients were divided into a none or tolerable pulmonary toxicity (NoSTox) group (grade ≤2) and a severe pulmonary toxicity (STox) group (grade = 3). Variant enrichment analysis allowed us to identify different pathway signatures characterizing NoSTox and Stox patients, allowing to formulate hypotheses on the protection from side effects derived from radiotherapy as well as factors predisposing to a worst response to the treatment. Our findings, being aware of the small number of patients analyzed, could be considered a starting point for the definition of a panel of pathways, possibly helpful in the management of MPM patients