Identification of biomarkers of meat tenderisation and its use for early classification of Asturian beef into fast and late tenderising meat.

BACKGROUND: The objective of this work was to study the post-mortem evolution of potential biomarkers (µ-calpain activity and proteolytic profile) of meat tenderisation in bovine longissimus dorsi (LD) muscle from several biotypes coming from two beef breeds ('Asturiana de los Valles' and 'Asturiana de la Montaña') and showing different levels of muscular hypertrophy (mh/mh, mh/+, + /+). RESULTS: LD samples were taken at 2, 12, 24 and 48 h and 3, 7, 14 and 21 days post-mortem. The presence of muscular hypertrophy produced a faster rate of pH decline, faster exhaustion of µ-calpain activity and earlier occurrence of proteolytic changes. Changes in the electrophoretic pattern of some peptides from sarcoplasmic (glyceraldehyde-3-phosphate dehydrogenase) and myofibrillar (troponin T and troponin I) muscle extracts within the first 24 h significantly correlated with meat toughness and allowed accurate discrimination of meat products into two groups: (1) fast tenderising meat, coming from mh-biotypes, and (2) late tenderising meat, from normal (+/+) biotypes. CONCLUSION: Early monitoring (within 24 h after slaughter) of selected biomarkers in LD muscle allowed accurate prediction of ultimate meat toughness and could be used in the meat industry as a tool for early classification of beef into fast and late tenderising meat.

Melatonin modulates autophagy through a redox-mediated action in female Syrian hamster Harderian gland controlling cell types and gland activity.

The Syrian hamster Harderian gland exhibits sexually dimorphic porphyrin biosynthesis, wherein the female glands display an extraordinarily high concentration of porphyrins. Damage derived from this production of porphyrins, mediated by reactive oxygen species, causes the glands to develop autophagic processes, which culminate in detachment-derived cell death; these cells normally play a central role in the secretory activity of the gland. The main aim of this study was to analyze how a change in the redox state impacts autophagy. Female Syrian hamsters were treated daily with melatonin (25 µg, subcutaneously) at ZT 10 for 1-2 months (N-acetyl-5-methoxytryptamine), an endogenous antioxidant that ameliorates the deleterious effects of free radicals via a variety of mechanisms. The length of treatment affected the redox balance, the autophagy machinery, and the activation of p53 and NF-κB. One-month treatment displaces redox balance to the antioxidant side, promotes autophagy through a p53-mediated mechanism, and increases cell detachment. Meanwhile, 2-month treatment restores redox balance to the oxidant side, activates NF-κB reducing autophagy to basal levels, increases number of type II cells, and reduces number of detached cells. Our results conclude that the redox state can modulate autophagy through redox-sensitive transcriptions factors. Additionally, these findings support a hypothesis that ascribes differences in the autophagic-lysosomal pathway to epithelial cell types, thereby restricting detachment-induced autophagic cell death to epithelial cell type I.

Sexual dimorphism of autophagy in Syrian hamster Harderian gland culminates in a holocrine secretion in female glands.

The Syrian hamster Harderian gland (HG) has a large porphyrin metabolism with a sexual dimorphism, showing male HGs much lower porphyrin concentrations than female glands. Damage derived from this production of porphyrins, displayed by reactive oxygen species, forces the gland to develop morphological changes that must have a physiological significance. Thus, oxidative stress is present in two states: mild oxidative stress in male HGs and extreme oxidative stress in female HGs. Cathepsins data gave indirect indications about the presence of programmed cell death affecting the lysosomal pathway, especially in female HGs, which showed an accumulation of autophagic bodies. Our results showed different degrees of autophagy in Syrian hamster HGs depending on sex and probably controlled by the redox-sensitive transcription factors: NFkappaB and p53. The discovery of these sexual dimorphisms in redox signaling and in autophagy corroborates previous findings and underlines the key role of reactive oxygen species in the regulation of autophagy. In addition, in this paper we propose a physiological significance for these phenomena: male HGs develop a survival autophagy, while in female HGs, autophagy culminates in a detachment-derived cell death that plays a central role in its secretory activity, leading to a massive glandular secretion.

Melatonin alters cell death processes in response to age-related oxidative stress in the brain of senescence-accelerated mice.

We studied the effect of age and melatonin on cell death processes in brain aging. Senescence-accelerated prone mice 8 (SAMP8) and senescence-accelerated resistant mice (SAMR1) at 5 and 10 months of age were used as models of the study. Melatonin (10 mg/kg) or its vehicle (ethanol at 0.066%) was administered in the drinking water from 1 to 9 months of age. Neurodegeneration, previously shown in the aged brain of SAMP8 and SAMR1 at 10 months of age, may be due to a drop in age-related proteolytic activities (cathepsin D, calpains, and caspase-3). Likewise, lack of apoptotic and macroautophagic processes were found, without apparent modification by melatonin. However, the caspase-independent cell death, owing to high p53 and apoptosis-inducing factor (AIF) levels, might be an alternative pathway of cell death in the aged brain. The main effects of melatonin treatment were observed in the aged SAMR1 mice; in this strain we observed a marked increase in antioxidant activity (catalase and superoxide dismutase). Likewise, a key antioxidant role of apoptosis-related proteins, Bcl-2 and AIF, was suggested in the aged brain of SAM mice, which was clearly influenced by melatonin. Moreover, the age-related increase of lysosomal activity of cathepsin B and a lysosomal membrane-associated protein 2 supports the possibility of the maintenance of lysosomal viability in addition to age-related impairments of the proteolytic or macroautophagic activities. The effectiveness of melatonin against the oxidative stress-related impairments and apoptosis during the aging process is, once more, corroborated in this article.

Precisión de la tomografía de 64 cortes en el diagnóstico de la cardiopatía isquémica / Accuracy of 64 slice computed tomography angiography in the diagnosis of ischemic coronary artery disease

INTRODUCTION AND OBJECTIVES: Multislice computed tomography coronary angiography (MSCT-CA) has been developed in the last years. One of the advantages is to supply information of the lumen and wall of the vessels. The aim was to assess the diagnostic accuracy of MSCT - CA to detect significant coronary stenoses taking as gold standard the invasive coronary angiography (ICA). PATIENTS AND METHODS: We studied, after informed consent, 64 consecutive patients (50 males). First MSCT - CA was performed and afterwards with a media of 45 days the ICA. Sensitivity (SENS), specificity (ESP), positive predictive value (PPV), negative predictive value (NPV) and predictive precision (PP) were assessed per patients, per arteries and per segments. RESULTS: The SENS, ESP, PPV, PNV and PP were 96.4, 91.2, 87, 96.8, 93.5% respectively per patients and 95.7, 97, 88.2, 98.9, 96.7% respectively per arteries and 91, 99, 89, 99.6, 98.7% respectively per segments. The diagnostic accuracy decreased in patients with heart rate higher than 65 beats per minute, as well as in patients with calcium scoring higher than 400 Agatston Units or body mass index with or higher than 30 kg/m2 CS. CONCLUSIONS: Our results suggest that MSCT - CA has a good accuracy, especially in the ESP and NPV for the diagnosis of significant coronary stenoses in selected patients with calcium scoring of 400 Agatston Units or lower, heart rate with 65 beats per minute or lower with regular rhythm and body mass index lower than 30 kg/m2 CS.

Precisión de la tomografía de 64 cortes en el diagnóstico de estenosis coronaria significativa / Accuracy of 64-slice computed tomography in the diagnosis of significant coronary stenosis

La coronariografía invasiva (CI) constituye el patrón de referencia para el estudio de las arterias coronarias. Se ha planteado que la tomografía computarizada de múltiples cortes (TCMC) puede evitar la CI. Se estudiaron 62 pacientes, 50 del sexo masculino, edad media 56 ± 8 años para evaluar la precisión diagnóstica de la TCMC de 64 cortes en la detección de estenosis coronarias significativas (ECS), comparándola con la CI. Se les realizó la CI por presentar ECS o persistencia de los síntomas. Se determinó sensibilidad (S), especificidad (E), valor predictivo positivo (VPP), valor predictivo negativo (VPN) y precisión predictiva (PP) por pacientes y por arterias. La S, la E, el VPP, el VPN y la PP fueron 96,4; 91,2; 87; 96,8 y 93,5 por ciento, respectivamente, por pacientes, y 95,7; 97; 88,2; 98,9 y 96,7 por ciento, respectivamente, por arterias. Se concluyó que la TCMC puede sustituir la CI en pacientes seleccionados.

Invasive coronariography (IC) is the reference pattern for the study of coronary arteries. It has been stated that multiple slice computed tomography (MSCT) may avoid IC. 62 patients, 50 of whom were males, with mean age 56 ± 8, were studied to evaluate the diagnostic accuracy of the 64-slice MSCT in the detection of significant coronary stenoses (SCS), comparing it with IC. IC was performed due to the presence of SCS or persistence of the symptoms. Sensitivity (S), specificity (E), positive predictive value (PPV), negative predictive value (NPV) and predictive accuracy (PA) were determined by patients and arteries. S, E, PPV, NPV and PA were 96.4; 91.2; 87; 96.8 and 93.5 percent per patient, respectively; whereas they were 95.7; 97; 88.2; 98.9 and 96.7 percent by artery, respectively. It was concluded that MSCT may replace IC in selected patients.

Coronariografía por tomografía de 64 cortes: Precisión diagnóstica según puntaje de calcio y frecuencia cardíaca / Coronariography by 64-Slice Computed Tomography: Diagnostic Accuracy According to Calcium Score and Heart Rate

Introducción y Objetivos La coronariografía invasiva (CI) es el patrón estándar para el estudio de las coronarias con un bajo índice de complicaciones. La tomografía computarizada multicorte (TCMC) permite el estudio mínimamente invasivo del árbol coronario. El objetivo del presente trabajo estuvo dirigido a determinar la precisión de la TCMC para detectar estenosis coronaria significativa (ECS) según puntaje de calcio y frecuencia cardíaca. Material y métodos Se estudiaron 62 pacientes, 50 del sexo masculino, a los que se les realizó primero la coronariografía por TCMC y luego la CI, con una media de 45 días, previo consentimiento informado. Se determinaron la sensibilidad (S), la especificidad (E), el valor predictivo positivo (VPP), el valor predictivo negativo (VPN) y la precisión predictiva (PP) por paciente y arterias según puntaje de calcio y frecuencia cardíaca. Resultados La S, la E, el VPP, el VPN y la PP por arterias fueron, respectivamente, con puntaje de calcio menor o igual a 400 UA del 98%, 98%, 89%, 99% y 98% y con puntaje mayor de 400 UA, del 75%, 50%, 75%, 50% y 67%. En los 54 pacientes con frecuencia cardíaca menor o igual a 65 latidos por minuto, la S, la E, el VPP, el VPN y la PP fueron del 100%, 96%, 96%, 100% y 98,1% y en los 8 pacientes con frecuencia cardíaca mayor de 65 latidos por minuto fueron del 50%, 66,7%, 33%, 80% y 62,5%, respectivamente. Conclusiones El puntaje de calcio por encima de 400 UA por paciente y arterias, como también la frecuencia cardíaca mayor de 65 latidos por minuto, disminuyen la precisión diagnóstica de ECS a través de la TCMC.

Introduction and Objectives Invasive coronariography (IC) is the gold standard for the study of coronary arteries with a low percentage of complications. Multislice computerized tomography (MSCT) allows the minimally invasive study of the coronary tree. The objective of the present study was aimed at determining the accuracy of MSCT in the detection of significant coronary stenosis (SCS) by calcium score and heart rate. Material and Methods The 62 patients enrolled in the study (50 males) underwent a coronariography with MSCT, and later the IC, with a mean of 45 days, after signature of an informed consent. Sensitivity (Se) was assessed, as well as specificity (Sp), positive predictive value (PPV), negative predictive value (NPV), and predictive accuracy (PA) per patient and arteries, according to calcium score and heart rate. Results Se, Sp, PPV, NPV and PA per arteries, with a calcium score lower or equal to 400 AU were, respectively 98%, 98%, 89%, 99% and 98%, and with a score higher than 400 AU, of 75%, 50%, 75%, 50% and 67%. In 54 patients with heart rate lower or equal to 65 beats per minute, Se, Sp, PPV, NPV and PA were 100%, 96%, 96%, 100% and 98,1% and in the 8 patients with heart rate higher than 65 beats per minute were 50%, 66.7%, 33%, 80% and 62.5%, respectively. Conclusions Calcium score above 400 AU per patient and arteries, as well as the heart rate over 65 beats per minute, decrease the diagnostic accuracy of SCS by means of MSCT.

Survival mechanisms in a physiological oxidative stress model.

The Syrian hamster Harderian gland has as the remarkable feature of an extraordinary rate of porphyrin production, even higher than the liver. The low activity of the last enzyme of the route gives rise to the accumulation of the uncomplex porphyrins in the female glands. Moreover, due to the localization of the Harderian gland, porphyrins exposed to light produce reactive oxygen species and, thus, the gland presents a physiological oxidative stress, with a great number of sings of degeneration, but without compromising the gland integrity. The appearance of abnormal features in this gland was largely described in the past, but the significance is interpreted for the first time in this study. We have found that autophagic processes are the first result of an elevated porphyrin metabolism, as it is observed in both sexes. This mechanism is considered, in this case, as a constant renovation system that allows the normal gland activity to be sustained. Furthermore, there is a second procedure, invasive processes toward connective tissue, which even occasionally reach blood vessels with intravasation of damaged gland components into the bloodstream. This effect is a consequence of a strong oxidative stress environment that is mainly observed in the female gland, resembling to tumoral progression. Both mechanisms, autophagy and invasive processes, have to be implied in the maintenance of the gland integrity.