Verónica Sierra

Favorable effects of a prolonged treatment with melatonin on the level of oxidative damage and neurodegeneration in senescence-accelerated mice

Abstract:  Senescence‐accelerated mice (SAMP8) and senescence‐accelerated resistant mice (SAMR1) were studied at 5 and 10 months of age, respectively. In the animals, neurodegenerative processes and how they were influenced by melatonin were examined. Melatonin (10 mg/kg) or vehicle (ethanol at 0.066%) treatments were administrated from the age of 1 to 9 months in the drinking water. Differences in the neurodegenerative markers examined were found between the two strains with a more damaged protein, phosphorylated Tau at Ser392, increased neurofibrillary tangles (NT) and higher α‐synuclein expression in SAMP8 versus SAMR1 mice overall, when the mice were 10 months of age. Changes in density of receptors and oxidative stress‐related signaling with age were found in the brains of SAM strains at 10 months as shown by a marked decrease in the level of MT‐1 melatonin receptor and retinoic acid receptor‐related orphan receptor (ROR)‐α1. This diminution was earlier and more pronounced in SAMP8 mice. Likewise, the levels of nuclear factor‐kappa B (NF‐kB) transcriptional factor were higher in SAMP8 mice compared with SAMR1 mice regardless of age confirming the direct role of oxidative stress in the aging process. Treatment with melatonin in SAMP8 and SAMR1 mice reduced the neurodegenerative changes with an increase of ROR‐α1 levels without an apparent influence in the levels of MT‐1 receptor. However, different melatonin effects on NF‐kB signaling were observed suggesting that NF‐kB could trigger inflammatory processes in a different way, being SAM strain‐dependent and associated with age‐related oxidative stress levels. The effectiveness of melatonin in improving age‐related neural impairments is corroborated.

Prediction of the fatty acid composition of beef by near infrared transmittance spectroscopy

The intramuscular fat content and composition influence consumer selection of meat products. A study predicting the fatty acid (FA) profile of ground beef from the Longissimus thoracis of yearling bulls (n=100) using near infrared transmittance spectroscopy (NIT) was conducted. The samples were scanned using an Infratec 1265 Meat Analyzer which operates in transmittance mode from 850 to 1050nm. NIT technology was able to accurately predict (R(CV)(2) over 0.76) some prominent FAs such as C14:0, C16:0, C16:1cis9, C17:0, C18:1cis9 and C18:1cis11, and minor FAs like C13:0, C15:0, C17:1cis9 and C18:1cis13. When studying FA groups, NIT spectra were able to accurately predict saturated (R(CV)(2)=0.837), branched (R(CV)(2)=0.701) and monounsaturated (R(CV)(2)=0.852) FAs. In addition, NIT spectra provided useful information on the contents of conjugated linoleic acids (CLA) in beef. These results show the potential of NIT technique as a rapid and easy tool to predict the major FAs in beef, especially those located in triglycerides.

Coexpression of MT1 and RORα1 melatonin receptors in the Syrian hamster Harderian gland

Abstract:  Melatonin acts through several specific receptors, including membrane receptors (MT1 and MT2) and members of the RZR/ROR nuclear receptors family, which have been identified in a large variety of mammalian and nonmammalian cells types. Both membrane and nuclear melatonin receptors have been partially characterized in Harderian gland of the Syrian hamster. Nevertheless, the identities of these receptors were unknown until this study, where the coexistence of MT1 and RORα1 in this gland was determined by nested RT‐PCR followed by amplicon sequencing and Western‐blot. Furthermore, the cellular localization of both receptors was determined by immunohistochemistry. Thus, MT1 receptor was localized exclusively at the basal side of the cell acini, supporting the hypothesis that this receptor is activated by the pineal‐synthesized melatonin. On the contrary, although a RORα1‐immunoreactivity was observed in nuclei of epithelial cells of both sexes, an extranuclear specific staining, which was more frequently among those cells of males, was also seen. The implication of this possible nuclear exclusion of RORα1 on the role of this indoleamine against oxidative stress is discussed.

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

Abstract:  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.

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

Abstract:  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.

Autophagy upregulation and loss of NF-κB in oxidative stress-related immunodeficient SAMP8 mice

Aged spleens from senescence-accelerated prone mice 8 (SAMP8) and senescence-accelerated resistant mice 1 (SAMR1) were examined to determine whether sex or melatonin had an effect on oxidative stress-related immune impairments. We observed that the immunosenescence of SAMP8 mice was associated with a redox imbalance, leading to an age-related increase in oxidative damage, resulting from a decrease in antioxidant defense and protease activity. Moreover, increased apoptotic cell death, a decrease in proliferative activity and the loss of NF-kappaB activation were also related to the immunodeficiency seen in SAMP8 compared to SAMR1 mice. Females demonstrated higher oxidative stress-related alterations in the immune response, and subsequent, melatonin treatment provided the best protective effects. Pathways involved in autophagy were upregulated in SAMP8 as an adaptive response to oxidative stress, in an attempt to rescue the cell from increased apoptosis and age-related immunodeficiency. However, the NF-kappaB signaling and autophagic processes were unaffected by treatment with melatonin. Therefore, we propose a key role for NF-kappaB signaling and autophagy in the oxidative stress-related immunosenescent spleens of SAMP8 mice.

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: NF-kappaB 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.

Antioxidant activity in Spalax ehrenbergi: a possible adaptation to underground stress

The blind subterranean mole rat Spalax ehrenbergi superspecies has evolved adaptive strategies to cope with underground stress. Hypoxia is known to stimulate reactive oxygen species generation; however, mechanisms by which Spalax counteracts oxidative damage have not been investigated before. We studied in Spalax the oxidative status of the Harderian gland (HG), an organ which is particularly vulnerable to oxidative stress in many rodents. With regard to the sexual dimorphism found in this gland, differences between males and females were determined and compared to the surface-dwelling Syrian hamster. Our results show, for the first time, that Spalax exhibits remarkably low biomolecular damage, which implies the existence of physiological strategies to avoid oxidative damage under fluctuating O2 and CO2 levels existing in the mole rat’s subterranean niche. Correspondingly, main antioxidant enzymes, such as superoxide dismutase (SOD), catalase, and glutathione reductase (GR), exhibited high activities in both genders; in particular, remarkably high levels were measured in SOD. SOD and GR activities showed statistically significant differences between sexes. Melatonin, an important circadian agent is also a very important antioxidant molecule and is synthesized in the Harderian glands (HGs) of Spalax. Therefore, the possible interaction between antioxidant enzymes and melatonin is suggested.

Systems Biology: A New Tool for Farm Animal Science

It is rapidly emerging that the tender meat phenotype is affected by an enormous amount of variables, not only tied to genetics (livestock breeding selection), but also to extrinsic factors, such as feeding conditions, physical activity, rearing environment, administration of hormonal growth promotants, pre-slaughter handling and stress. Proteomics has been widely accepted by meat scientists over the last years and is now commonly used to shed light on the postmortem processes involved in meat tenderization. This review discusses the latest findings with the use of proteomics and systems biology to study the different biochemical pathways postmortem aiming at understanding the concerted action of different molecular mechanisms responsible for meat quality. The conversion of muscle to meat postmortem can be described as a sequence of events involving molecular pathways controlled by a complex interplay of many factors. Among the different pathways emerging are the influence of apoptosis and lately also the role of autophagy in muscle postmortem development. This review thus, focus on how systems-wide integrated investigations (metabolomics, transcriptomics, interactomics, phosphoproteomics, mathematical modeling), which have emerged as complementary tools to proteomics, have helped establishing a few milestones in our understanding of the events leading from muscle to meat conversion.

Sexual Autophagic Differences in the Androgen-Dependent Flank Organ of Syrian Hamsters

The flank organ of the Syrian hamster shows a biodynamic response to androgenic stimulation and is, therefore, a suitable model for the study of androgenic effects on hair and sebaceous glands. This organ is susceptible to programmed cell death (PCD), a prominent feature associated with sexual organ adjustment. In the present report, the type of PCD (apoptosis or autophagy) exhibited by this organ was evaluated. Caspase-3 activity, indicative of apoptosis, was not detectable in flank organ homogenates. Furthermore, cytokeratins, which are normally degraded during apoptosis, remained intact. On the other hand, Western blotting of Beclin 1 and light chain 3-II, both important autophagy markers, revealed autophagic processes in the flank organ in both sexes, especially in females. Cathepsin D activity, higher in males than in females, and procathepsin D expression were also consistent with autophagy and not apoptosis. Taken together, these data indicate that macroautophagy, and not apoptosis, is the main mechanism by which the flank organ responds to androgen. This is the first direct evidence establishing the relationship between autophagy and morphological changes in androgen-dependent organs.