Evolution and treatment of vitamin B12 deficiency as a risk factor for (cognitive and functional) neurodegenerative diseases in institutionalized elderly = Evolución y tratamiento de la deficiencia de vitamina B12 como factor de riesgo de enfermedades neurodegenerativas (cognitivas y funcionales) en las personas mayores institucionalizadas.

Prevalence of vitamin B12 deficiency is very common in elderly people and can reach values as high as 40.5% of the population. It can be the result of the interaction among several factors. Vitamin B12 deficiencies have been associated with neurological, cognitive deterioration, haematological abnormalities and cardiovascular diseases that have an important influence on the health of the elderly and their quality of life. It is necessary to approach the problems arisen from the lack of data relative to them. The main objective of this thesis was to analyse the evolution of vitamin B12 status and related parameters, lipid and haematological profiles and their relationship to health risk factors, and to functional and cognitive status over one year and to determine the effect of an oral supplementation of 500 μg of cyanocobalamin for a short period of 28 days. An additional objective was to analyze the possible effects of medicine intakes on vitamin B status. Three studies were performed: a) a one year longitudinal follow-up with four measure points; b) an intervention study providing an oral liquid supplement of 500 μg of cyanocobalamin for a 28 days period; and c) analysis of the possible effect of medication intake on vitamin B status using the ATC classification of medicines. The participants for these studies were recruited from nursing homes for the elderly in the Region of Madrid. Sixty elders (mean age 84 _ 7y, 19 men and 41 women) were recruited for Study I and 64 elders (mean age 82 _ 7y, 24 men and 40 women) for Study II. For Study III, baseline data from the initially recruited participants of the first two studies were used. An informed consent was obtained from all participants or their mentors. The studies were approved by the Ethical Committee of the University of Granada. Blood samples were obtained at each examination date and were analyzed for serum cobalamin, holoTC, serum and RBC folate and total homocysteine according to laboratory standard procedures. The haematological parameters analyzed were haematocrit, haemoglobin and MCV. For the lipid profile TG, total cholesterol, LDL- and HDLcholesterol were analyzed. Anthropometric measures (BMI, skinfolds [triceps and subscapular], waist girth and waist to hip ratio), functional tests (hand grip, arm and leg strength tests, static balance) and MMSE were obtained or administered by trained personal. The vitamin B12 supplement of Study II was administered with breakfast and the medication intake was taken from the residents’ anamnesis. Data were analyzed by parametric and non-parametric statistics depending on the obtained data. Comparisons were done using the appropriate ANOVAs or non-parametric tests. Pearsons’ partial correlations with the variable “time” as control were used to define the association of the analyzed parameters. XIII The results showed that: A) Over one year, in relationship to vitamin B status, serum cobalamin decreased, serum folate and mean corpuscular volumen increased significantly and total homocysteine concentrations were stable. Regarding blood lipid profile, triglycerides increased and HDL-cholesterol decreased significantly. Regarding selected anthropometric measurements, waist circumference increased significantly. No significant changes were observed for the rest of parameters. B) Prevalence of hyperhomocysteinemia was high in the elderly studied, ranging from 60% to 90 % over the year depending on the cut-off used for the classification. LDL-cholesterol values were high, especially among women, and showed a tendency to increase over the year. Results of the balance test showed a deficiency and a tendency to decrease; this indicates that the population studied is at high risk for falls. Lower extremity muscular function was deficient and showed a tendency to decrease. A highly significant relationship was observed between the skinfold of the triceps and blood lipid profile. C) Low cobalamin concentrations correlated significantly with low MMSE scores in the elderly studied. No correlations were observed between vitamin B12 status and functional parameters. D) Regarding vitamin B12 status, holo-transcobalamin seems to be more sensitive for diagnosis; 5-10% of the elderly had a deficiency using serum cobalamin as a criterion, and 45-52% had a deficiency when using serum holotranscobalamin as a criterion. E) 500 μg of cyanocobalamin administered orally during 28 days significantly improved vitamin B12 status and significantly decreased total homocysteine concentrations in institutionalized elderly. No effect of the intervention was observed on functional and cognitive parameters. F) The relative change (%) of improvement of vitamin B12 status was higher when using serum holo-transcobalamin as a criterion than serum cobalamin. G) Antiaenemic drug intake normalized cobalamin, urologic drugs and corticosteroids serum folate, and psychoanaleptics holo-transcobalamin levels. Drugs treating pulmonary obstruction increased total homocysteine concentration significantly. H) The daily mean drug intake was 5.1. Fiftynine percent of the elderly took medication belonging to 5 or more different ATC groups. The most prevalent were psycholeptic (53%), antiacid (53%) and antithrombotic (47%) drugs.

Effect of short-term fasting prior to slaughter on welfare and meat quality or rainbow trout (Oncorhynchus mykiss)

El manejo pre-sacrificio es de vital importancia en acuicultura, ya que afecta tanto a las reacciones fisiológicas como a los procesos bioquímicos post mortem, y por tanto al bienestar y a la calidad del producto. El ayuno pre-sacrificio se lleva a cabo de forma habitual en acuicultura, ya que permite el vaciado del aparato digestivo de restos de alimento y heces, reduciendo de esta manera la carga bacteriana en el intestino y la dispersión de enzimas digestivos y potenciales patógenos a la carne. Sin embargo, la duración óptima de este ayuno sin que el pez sufra un estrés innecesario no está clara. Además, se sabe muy poco sobre la mejor hora del día para realizar el sacrificio, lo que a su vez está regido por los ritmos diarios de los parámetros fisiológicos de estrés. Finalmente, se sabe que la temperatura del agua juega un papel muy importante en la fisiología del estrés pero no se ha determinado su efecto en combinación con el ayuno. Además, las actuales recomendaciones en relación a la duración óptima del ayuno previo al sacrificio en peces no suelen considerar la temperatura del agua y se basan únicamente en días y no en grados día (ºC d). Se determinó el efecto del ayuno previo al sacrificio (1, 2 y 3 días, equivalente a 11,1-68,0 grados día) y la hora de sacrificio (08h00, 14h00 y 20h00) en trucha arco iris (Oncorhynchus mykiss) de tamaño comercial en cuatro pruebas usando diferentes temperaturas de agua (Prueba 1: 11,8 ºC; Prueba 2: 19,2 ºC; Prueba 3: 11,1 ºC; y Prueba 4: 22,7 ºC). Se midieron indicadores biométricos, hematológicos, metabólicos y de calidad de la carne. En cada prueba, los valores de los animales ayunados (n=90) se compararon con 90 animales control mantenidos bajo condiciones similares pero nos ayunados. Los resultados sugieren que el ayuno tuvo un efecto significativo sobre los indicadores biométricos. El coeficiente de condición en los animales ayunados fue menor que en los controles después de 2 días de ayuno. El vaciado del aparato digestivo se produjo durante las primeras 24 h de ayuno, encontrándose pequeñas cantidades de alimento después de 48 h. Por otra parte, este vaciado fue más rápido cuando las temperaturas fueron más altas. El peso del hígado de los animales ayunados fue menor y las diferencias entre truchas ayunadas y controles fueron más evidentes a medida que el vaciado del aparato digestivo fue más rápido. El efecto del ayuno hasta 3 días en los indicadores hematológicos no fue significativo. Los niveles de cortisol en plasma resultaron ser altos tanto en truchas ayunadas como en las alimentadas en todas las pruebas realizadas. La concentración media de glucosa varió entre pruebas pero mostró una tendencia a disminuir en animales ayunados a medida que el ayuno progresaba. En cualquier caso, parece que la temperatura del agua jugó un papel muy importante, ya que se encontraron concentraciones más altas durante los días 2 y 3 de ayuno en animales mantenidos a temperaturas más bajas previamente al sacrificio. Los altos niveles de lactato obtenidos en sangre parecen sugerir episodios de intensa actividad muscular pero no se pudo encontrar relación con el ayuno. De la misma manera, el nivel de hematocrito no mostró efecto alguno del ayuno y los leucocitos tendieron a ser más altos cuando los animales estaban menos estresados y cuando su condición corporal fue mayor. Finalmente, la disminución del peso del hígado (índice hepatosomático) en la Prueba 3 no se vio acompañada de una reducción del glucógeno hepático, lo que sugiere que las truchas emplearon una estrategia diferente para mantener constantes los niveles de glucosa durante el periodo de ayuno en esa prueba. En relación a la hora de sacrificio, se obtuvieron niveles más bajos de cortisol a las 20h00, lo que indica que las truchas estaban menos estresadas y que el manejo pre-sacrificio podría resultar menos estresante por la noche. Los niveles de hematocrito fueron también más bajos a las 20h00 pero solo con temperaturas más bajas, sugiriendo que las altas temperaturas incrementan el metabolismo. Ni el ayuno ni la hora de sacrificio tuvieron un efecto significativo sobre la evolución de la calidad de la carne durante los 3 días de almacenamiento. Por el contrario, el tiempo de almacenamiento sí que parece tener un efecto claro sobre los parámetros de calidad del producto final. Los niveles más bajos de pH se alcanzaron a las 24-48 h post mortem, con una lata variabilidad entre duraciones del ayuno (1, 2 y 3 días) en animales sacrificados a las 20h00, aunque no se pudo distinguir ningún patrón común. Por otra parte, la mayor rigidez asociada al rigor mortis se produjo a las 24 h del sacrificio. La capacidad de retención de agua se mostró muy estable durante el período de almacenamiento y parece ser independiente de los cambios en el pH. El parámetro L* de color se incrementó a medida que avanzaba el período de almacenamiento de la carne, mientras que los valores a* y b* no variaron en gran medida. En conclusión, basándose en los resultados hematológicos, el sacrificio a última hora del día parece tener un efecto menos negativo en el bienestar. De manera general, nuestros resultados sugieren que la trucha arco iris puede soportar un período de ayuno previo al sacrificio de hasta 3 días o 68 ºC d sin que su bienestar se vea seriamente comprometido. Es probable que con temperaturas más bajas las truchas pudieran ser ayunadas durante más tiempo sin ningún efecto negativo sobre su bienestar. En cualquier caso, se necesitan más estudios para determinar la relación entre la temperatura del agua y la duración óptima del ayuno en términos de pérdida de peso vivo y la disminución de los niveles de glucosa en sangre y otros indicadores metabólicos. SUMMARY Pre-slaughter handling in fish is important because it affects both physiological reactions and post mortem biochemical processes, and thus welfare and product quality. Pre-slaughter fasting is regularly carried out in aquaculture, as it empties the viscera of food and faeces, thus reducing the intestinal bacteria load and the spread of gut enzymes and potential pathogens to the flesh. However, it is unclear how long rainbow trout can be fasted before suffering unnecessary stress. In addition, very little is known about the best time of the day to slaughter fish, which may in turn be dictated by diurnal rhythms in physiological stress parameters. Water temperature is also known to play a very important role in stress physiology in fish but the combined effect with fasting is unclear. Current recommendations regarding the optimal duration of pre-slaughter fasting do not normally consider water temperature and are only based on days, not degree days (ºC d). The effects of short-term fasting prior to slaughter (1, 2 and 3 days, between 11.1 and 68.0 ºC days) and hour of slaughter (08h00, 14h00 and 20h00) were determined in commercial-sized rainbow trout (Oncorhynchus mykiss) over four trials at different water temperatures (TRIAL 1, 11.8 ºC; TRIAL 2, 19.2 ºC; TRIAL 3, 11.1 ºC; and TRIAL 4, 22.7 ºC). We measured biometric, haematological, metabolic and product quality indicators. In each trial, the values of fasted fish (n=90) were compared with 90 control fish kept under similar conditions but not fasted. Results show that fasting affected biometric indicators. The coefficient of condition in fasted trout was lower than controls 2 days after food deprivation. Gut emptying occurred within the first 24 h after the cessation of feeding, with small traces of digesta after 48 h. Gut emptying was faster at higher water temperatures. Liver weight decreased in food deprived fish and differences between fasted and fed trout were more evident when gut clearance was faster. The overall effect of fasting for up to three days on haematological indicators was small. Plasma cortisol levels were high in both fasted and fed fish in all trials. Plasma glucose response to fasting varied among trials, but it tended to be lower in fasted fish as the days of fasting increased. In any case, it seems that water temperature played a more important role, with higher concentrations at lower temperatures on days 2 and 3 after the cessation of feeding. Plasma lactate levels indicate moments of high muscular activity and were also high, but no variation related to fasting could be found. Haematocrit did not show any significant effect of fasting, but leucocytes tended to be higher when trout were less stressed and when their body condition was higher. Finally, the loss of liver weight was not accompanied by a decrease in liver glycogen (only measured in TRIAL 3), suggesting that a different strategy to maintain plasma glucose levels was used. Regarding the hour of slaughter, lower cortisol levels were found at 20h00, suggesting that trout were less stressed later in the day and that pre-slaughter handling may be less stressful at night. Haematocrit levels were also lower at 20h00 but only at lower temperatures, indicating that higher temperatures increase metabolism. Neither fasting nor the hour of slaughter had a significant effect on the evolution of meat quality during 3 days of storage. In contrast, storage time seemed to have a more important effect on meat quality parameters. The lowest pH was reached 24-48 h post mortem, with a higher variability among fasting durations at 20h00, although no clear pattern could be discerned. Maximum stiffening from rigor mortis occurred after 24 h. The water holding capacity was very stable throughout storage and seemed to be independent of pH changes. Meat lightness (L*) slightly increased during storage and a* and b*-values were relatively stable. In conclusion, based on the haematological results, slaughtering at night may have less of a negative effect on welfare than at other times of the day. Overall, our results suggest that rainbow trout can cope well with fasting up to three days or 68 ºC d prior to slaughter and that their welfare is therefore not seriously compromised. At low water temperatures, trout could probably be fasted for longer periods without negative effects on welfare but more research is needed to determine the relationship between water temperature and days of fasting in terms of loss of live weight and the decrease in plasma glucose and other metabolic indicators.

Mouse p53-deficient cancer models as platforms for obtaining genomic predictors of human cancer clinical outcomes

Mutations in the TP53 gene are very common in human cancers, and are associated with poor clinical outcome. Transgenic mouse models lacking the Trp53 gene or that express mutant Trp53 transgenes produce tumours with malignant features in many organs. We previously showed the transcriptome of a p53-deficient mouse skin carcinoma model to be similar to those of human cancers with TP53 mutations and associated with poor clinical outcomes. This report shows that much of the 682-gene signature of this murine skin carcinoma transcriptome is also present in breast and lung cancer mouse models in which p53 is inhibited. Further, we report validated gene-expression-based tests for predicting the clinical outcome of human breast and lung adenocarcinoma. It was found that human patients with cancer could be stratified based on the similarity of their transcriptome with the mouse skin carcinoma 682-gene signature. The results also provide new targets for the treatment of p53-defective tumours.

Optical detection of mealiness in apples using laser TDRS.

Mealiness is a textural attribute related to an internal fruit disorder that involves quality loss. It is characterised by the combination of abnormal softness of the fruit and absence of free juiciness in the mouth when eaten by the consumer. Recent research concluded with the development of precise instrumental procedure to measure a scale of mealiness based on the combination of several rheological properties and empirical magnitudes. In this line, time-domain laser reflectance spectroscopy (TDRS) is a new medical technology, used to characterise the optical properties of tissues, and to locate affected areas like tumours. Among its advantages compared to more traditional spectroscopic techniques, there is the feasibility to asses simultaneously and independently two optical parameters: the absorption of the light inside the irradiated body, and the scattering of the photons across the tissues, at each wavelength, generating two coefficients (µa, absorption coeff.; and µ's, transport scattering coeff.). If it is assumed that they are related respectively to chemical components and to physical properties of the sample, TDRS can be applied to the quantification of chemicals and the measurement of the rheological properties (i.e. mealiness estimation) at the same time. Using VIS & NIR lasers as light sources, TDRS was applied in this work to Golden Delicious and Cox apples (n=90), conforming several batches of untreated samples and storage-treated (20°C & 95%RH) to promote the development of mealiness. The collected database was clustered into different groups according to their instrumental test values (Barreiro et al, 1998). The optical coefficients were used as explanatory variables when building discriminant analysis functions for mealiness, achieving a classification score above 80% of correctly identified mealy versus fresh apples.

Análisis de la estructura del juego y parámetros morfológicos y fisiológicos en bádminton

En este trabajo se ha realizado un análisis de la estructura del juego y de los parámetros morfológicos y fisiológicos en jugadores de bádminton. Para ello se han realizado 4 estudios aplicados. Objetivo: Los objetivos del trabajo han sido: (1) comprobar si existen diferencias entre el lado dominante y no dominante de las medidas antropométricas en jugadores de bádminton de máximo nivel nacional, así como verificar si el lado del cuerpo donde se realiza la medición puede influir en el cálculo de la composición corporal y del somatotipo. (2) Comparar la estuctura temporal y notacional en partidos de individual masculino entre los Juegos Olímpicos de Pekín y de Londres para observar como ha evolucionado el bádminton de 2008 a 2012. (3) Medir la ocurrencia de daño muscular después de un partido simulado de bádminton y su influencia en parámetros físicos y hematológicos. (4) Investigar la efectividad de una bebida energética que contiene cafeína para mejorar el rendimiento físico y el rendimiento en un partido en jugadores de élite de bádminton. Metodología: Para caracterizar el bádminton participaron en esta tesis un total de 78 jugadores de bádminton de élite (63 hombres y 15 mujeres), distribuidos en tres estudios y se analizaron 40 sets de bádminton de individual masculino usando los videos oficiales de los Juegos Olímpicos de Pekín 2008 y Londres 2012. En el primer estudio se tomaron medidas de pliegues cutáneos, diámetros, longitudes y perímetros del lado dominante y no dominante de los jugadores. Se calculó la composición corporal y el somatotipo. En el segundo estudio se analizaron los factores temporales y los factores notacionales de los partidos. En el tercer estudio se midieron la fuerza máxima isométrica, la velocidad en test específicos de bádminton y se tomaron muestras de sangre antes y después de jugar un partido de bádminton de 45 minutos. En el cuarto estudio se realizó un experimento a doble ciego, aleatorizado y controlado con placebo, los jugadores ingirieron 3 mg de cafeína por kilógramo de masa corporal en forma de bebida energética, o la misma bebida sin cafeína (placebo). En este estudio se registraron diferente tests específicos de bádminton (tests de salto, fuerza máxima y test de agilidad) y se jugó un partido simulado de 45 minutos. Resultados y discusión: (1) El porcentaje óseo fue mayor calculado a partir de las mediciones del lado dominante (dominante = 16.37 ± 1.14 %, no dominante = 15.66 ± 1.12 %; P < 0.001), mientras que el porcentaje muscular fue mayor calculado a partir de las mediciones del lado no dominante (dominante = 49.39 ± 2.60 %, no dominante = 50.18 ± 2.69%; P < 0.001). (2) La duración del set (Pekín: 1124.6 ± 229.9 s vs Londres: 1260.3 ± 267.1 s.; P < 0.05), el tiempo real de juego (Pekín: 306.9 ± 45.7 s vs Londres: 354.7 ± 86.5 s; P < 0.05), tiempo de rally, golpeos por rally, tiempo de descanso en el punto 11, tiempo de descanso entre sets y golpeos por rally fueron significativamente mayores en Londres que en Pekín. (3) El partido simulado de bádminton no afectó a la fuerza isométrica máxima (Pre: 1263.6 ± 245.5, Post: 1290.8 ± 240.4 N) o a la velocidad específica de bádminton (Pre: 21.0 ± 1.7, Post: 20.9 ± 1.8 s), sin embargo las concentraciones de mioglobina y de creatina quinasa en sangre aumentaron de 26.5 ± 11.6 a 197.3 ± 70.2 μg • L-1 y de 258.6 ± 192.2 a 466.0 ± 296.5 U • L-1, respectivamente después del partido de bádminton. (4) En comparación con la bebida placebo, la ingesta de la bebida energética con cafeína incrementó la altura del SJ (34.5±4.7 vs. 36.4±4.3 cm; P < 0.05) y del CMJ (37.7 ± 4.5 vs. 39.5 ± 5.1 cm; P < 0.05) y aumentó el número de aceleraciones totales durante el partido (7395 ± 1594 vs. 7707 ± 2033 aceleraciones; P < 0.05). Conclusiones: (1) Existen asimetrías corporales en los jugadores de bádminton de alto nivel, al encontrarse diferencias en los diámetros óseos y en los perímetros entre el lado dominante y no dominante. Al calcular la composición corporal con el lado dominante de los jugadores de bádminton se está sobreestimando el porcentaje óseo e infraestimando el porcentaje muscular. (2) El bádminton está evolucionando hacía rallies más largos con intervalos de descanso mayores, lo que resulta en partidos más largos. (3) El partido de bádminton generó daño muscular, sin embargo, el nivel de daño muscular alcanzado después de un partido de bádminton no produjo una disminución del rendimiento muscular. (4) El uso de una bebida energética con cafeína puede ser una ayuda nutricional eficaz para aumentar el rendimiento en el salto y patrones de actividad durante el juego en jugadores de élite de bádminton. ABSTRACT: This study analyzes the structure of the game and the morphological and physiological parameters in badminton players, investigated in four applied studies. Purpose: The purposes of the study were: (1) To check if there are differences between the dominant and non-dominant side in the anthropometric measures of badminton players at the highest national level and verify if the side of the body where the measurements are performed can influence the calculation of the body composition and the somatotype. (2) To compare the temporal and notational structure in men’s singles matches between the Olympic Games in Beijing and London to observe the evolution of badminton between 2008 and 2012. (3) To asses the occurrence of muscle damage after a simulated badminton match and its influence on physical and haematological parameters. (4) To determine the effectiveness of a commercially available energy drink that contains caffeine to improve match performance in elite badminton players. Methods: A total of 78 elite badminton players (63 men and 15 women) participated in this thesis to characterize the sport of badminton distributed in three studies and 40 sets of men’s singles badminton analyzed using the official videos of the Olympic Games of Beijing 2008 and London 2012. In the first study skinfolds, diameters, lengths and perimeters of the dominant and non-dominant side of the players were measured and body composition and somatotype were calculated. In the second study the temporal and notational factors were analyzed. In the third study maximal isometric force and speed in badminton specific tests were measured and blood samples were taken before and after a badminton match of 45 minutes. In the fourth study, a double-blind, randomized placebo-controlled experiment, players ingested 3 mg of caffeine per kilogram of body mass in the form of an energy drink or an identical drink with no caffeine content (placebo). In this study different badminton specific tests (jump tests, handgrip force test and an agility test) were recorded and a simulated badminton match of 45 minutes was played. Results and discussion: (1) The percentage of bone was higher when calculated from measurements of the dominant body side (dominant = 16.37 ± 1.14 %, nondominant = 15.66 ± 1.12 %; P < 0.001), while the muscle percentage was higher when calculated from measurements of the non-dominant side (dominant = 49.39 ± 2.60 %, non-dominant = 50.18 ± 2.69%; P < 0.001). (2) Set duration (Beijing: 1124.6 ± 229.9 s vs. London: 1260.3 ± 267.1 s.; P < 0.05), real time played (Beijing: 306.9 ± 45.7 s vs. London: 354.7 ± 86.5 s; P < 0.05), rally time, shots per rally, rest time at point 11, rest time between sets and shots per rally were significantly higher in London than in Beijing. (3) A simulated badminton match did not affect maximal isometric force (Pre: 1263.6 ± 245.5, Post: 1290.8 ± 240.4 N) or specific badminton speed (Pre: 21.0 ± 1.7, Post: 20.9 ± 1.8 s), however, concentrations of myoglobin and creatine kinase in blood increased from 26.5 ± 11.6 to 197.3 ± 70.2 μg • L-1 and from 258.6 ± 192.2 to 466.0 ± 296.5 U • L-1, respectively after the badminton match. (4) In comparison to the placebo drink, the caffeinated beverage increased height in the SJ (34.5±4.7 vs. 36.4±4.3 cm; P < 0.05) and in the CMJ (37.7 ± 4.5 vs. 39.5 ± 5.1 cm; P < 0.05) and increased the number of total accelerations during the match (7395 ± 1594 vs. 7707 ± 2033 accelerations; P < 0.05). Conclusions: (1) Body asymmetries were found in high level badminton players, due to the differences found in bone diameters and perimeters between the dominant and non-dominant body side. When calculating body composition with the dominant side of the badminton players we are overestimating bone percentage and underestimating muscle percentage. (2) Badminton is evolving towards longer rallies with greater rest intervals, resulting in longer matches. (3) The badminton match generated muscle damage, however, the level of muscle damage reached after a badminton match did not produce a decrease in muscle performance. (4) The ingestion of an energy drink containing caffeine might be an effective ergogenic nutritional supplement to increase jump performance and activity patterns during the game in elite badminton players.