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Recent advances in "omics" technologies have enabled the identification of new beef quality biomarkers and have also allowed for the early detection of quality defects such as dark-cutting beef, also known as DFD (dark, firm, and dry) beef. However, most of the studies conducted were carried out on a small number of animals and mostly applied gel-based proteomics. The present study proposes for the first time a Sequential Window Acquisition of All Theoretical Mass Spectra (SWATH-MS) proteomics approach to characterize and comprehensively quantify the post-mortem muscle proteome of DFD (pH24 ≥ 6.2) and CONTROL (5.4 ≤ pH24 ≤ 5.6) beef samples within the largest database of DFD/CONTROL beef samples to date (26 pairs of the Longissimus thoracis muscle samples of young bulls from Asturiana de los Valles breed, n = 52). The pairwise comparison yielded 35 proteins that significantly differed in their abundances between the DFD and CONTROL samples. Chemometrics methods using both PLS-DA and OPLS-DA revealed 31 and 36 proteins with VIP > 2.0, respectively. The combination of different statistical methods these being Volcano plot, PLS-DA and OPLS-DA allowed us to propose 16 proteins as good candidate biomarkers of DFD beef. These proteins are associated with interconnected biochemical pathways related to energy metabolism (DHRS7B and CYB5R3), binding and signaling (RABGGTA, MIA3, BPIFA2B, CAP2, APOBEC2, UBE2V1, KIR2DL1), muscle contraction, structure and associated proteins (DMD, PFN2), proteolysis, hydrolases, and activity regulation (AGT, C4A, GLB1, CAND2), and calcium homeostasis (ANXA6). These results evidenced the potential of SWATH-MS and chemometrics to accurately identify novel biomarkers for meat quality defects, providing a deeper understanding of the molecular mechanisms underlying dark-cutting beef condition.
Assuntos
Biomarcadores , Músculo Esquelético , Proteômica , Carne Vermelha , Animais , Bovinos , Carne Vermelha/análise , Biomarcadores/análise , Proteômica/métodos , Masculino , Músculo Esquelético/química , Espectrometria de Massas/métodos , Proteoma/análise , Proteínas Musculares/análiseRESUMO
In an animal production system, different stressors may cause the depletion of muscle glycogen stores, resulting in an elevated pH at 24 h post mortem (pH24), which leads to cell metabolism alterations that affect the conversion of muscle into meat, causing meat quality defects, such as dark-cutting beef, also known as dark, firm, and dry (DFD) beef. This process may involve the alteration of small non-coding RNAs (miRNAs), which play critical regulatory roles in cellular processes. Here, we determined whether differential miRNA expression in the Longissimus thoracis et lumborum muscle from the Asturiana de los Valles breed at 24 h post mortem could serve as an early indicator of beef quality defects. Following total RNA extraction, complete miRNAome sequencing revealed 12 miRNAs that were significantly upregulated (p < 0.001) in DFD beef compared to the levels in CONTROL beef. These miRNAs are mainly involved in the cellular responses to redox imbalances and apoptosis. Among these, four miRNAs known to be related to oxidative stress (bta-miR-1246, bta-miR-2332, bta-miR-23b-5p, and bta-miR-2411-3p) were validated via quantitative RT-PCR. Some of their target proteins were also analyzed using Western blotting. High 70 kDa heat shock protein and low Caspase-9 expressions (p < 0.01) were found in DFD beef, suggesting the downregulation of apoptosis. These results suggest the importance of miRNAs in regulating stress in muscle cells during early post mortem, as differences in the abundance of some of these miRNAs are still observed at 24 h post mortem. These changes lead to an inadequate conversion of muscle into meat, resulting in meats with quality defects.
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For fresh meat consumers, eating satisfaction is of utmost importance and tenderness is one of the most important characteristics in this regard. Our study examined beef of different animal biotypes of the autochthonous breed "Asturiana de los Valles" (AV) to determine if early postmortem oxidative and proteolytic processes may influence the final tenderness of the product. This meat-specialized breed shows different biotypes depending on the frequency of a myostatin mutation "mh" that induces double-muscling or muscular hypertrophy (mh/mh, mh/+, +/+). Samples from the longissimus dorsi muscles of yearling bulls were analyzed during the first 24 h postmortem. Changes in the redox balance of muscle cells were significant in the first hours after slaughter; total antioxidant activity was higher in the mh/mh biotype and it followed the shortening of the sarcomeres, a key parameter in understanding meat tenderness. The two proteolytic systems studied (proteasome and lysosome) followed distinct patterns. Proteasome activity was higher in the (mh/+) biotype, which correlated with higher protein damage. Lysosome proteolysis was increased in the more tender biotypes (mh genotypes). Autophagic activation showed significant differences between the biotypes, with (mh/mh) showing more intense basal autophagy at the beginning of the postmortem period that decreased gradually (p < 0.001), while in the normal biotype (+/+), it was slightly delayed and then increased progressively (p < 0.001). These results suggest that this type of catalytic process and antioxidant activity could contribute to the earlier disintegration of the myofibers, particularly in the mh/mh biotypes, and influence the conversion of muscle into meat.
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Defects in meat quality such as dark, firm and dry (DFD) beef have been related to high levels of oxidative stress that produce cellular alterations that may affect to the process of meat quality acquisition. Despite the important role of endoplasmic reticulum (ER) in the cellular response to oxidative stress, its function in the muscle-to-meat conversion process has not yet been studied. In this study, differences in muscular antioxidant defense and the unfolded protein response (UPR) of the ER in CONTROL (normal pH24) and dark, firm, and dry (DFD, pH24 ≥ 6.2) beef at 24 h post-mortem were analyzed to understand the changes in the muscle-to-meat conversion process related to meat quality defects. DFD meat showed poor quality, lower antioxidant activity (P < 0.05) and higher UPR activation (P < 0.05), which indicates higher oxidative stress what could partly explain the occurrence of meat quality defects. Therefore, the biomarkers of these cellular processes (IRE1α, ATF6α, and p-eIF2α) are putative biomarkers of meat quality.
Assuntos
Endorribonucleases , Proteínas Serina-Treonina Quinases , Animais , Bovinos , Proteínas Serina-Treonina Quinases/metabolismo , Endorribonucleases/metabolismo , Retículo Endoplasmático/metabolismo , Carne , Estresse do Retículo EndoplasmáticoRESUMO
There are several neurological diseases under which processes related to adult brain neurogenesis, such cell proliferation, neural differentiation and neuronal maturation, are affected. Melatonin can exert a relevant benefit for treating neurological disorders, given its well-known antioxidant and anti-inflammatory properties as well as its pro-survival effects. In addition, melatonin is able to modulate cell proliferation and neural differentiation processes in neural stem/progenitor cells while improving neuronal maturation of neural precursor cells and newly created postmitotic neurons. Thus, melatonin shows relevant pro-neurogenic properties that may have benefits for neurological conditions associated with impairments in adult brain neurogenesis. For instance, the anti-aging properties of melatonin seem to be linked to its neurogenic properties. Modulation of neurogenesis by melatonin is beneficial under conditions of stress, anxiety and depression as well as for the ischemic brain or after a brain stroke. Pro-neurogenic actions of melatonin may also be beneficial for treating dementias, after a traumatic brain injury, and under conditions of epilepsy, schizophrenia and amyotrophic lateral sclerosis. Melatonin may represent a pro-neurogenic treatment effective for retarding the progression of neuropathology associated with Down syndrome. Finally, more studies are necessary to elucidate the benefits of melatonin treatments under brain disorders related to impairments in glucose and insulin homeostasis.
Assuntos
Melatonina , Células-Tronco Neurais , Melatonina/farmacologia , Hipocampo , Neurogênese , NeurôniosRESUMO
Real-time monitoring of meat quality requires fast, accurate, low-cost, and non-destructive analytical methods that can be used throughout the entire production chain, including the packaged product. The aim of this work was to evaluate the potential of a portable near-infrared (NIR) spectroscopy tool for the on-site detection of freshness of pork loin fillets in modified atmosphere packaging (MAP) stored on display counters. Pork loin slices were sealed in MAP trays under two proportions of O2/CO2/N2: High-Ox-MAP (30/40/30) and Low-Ox-MAP (5/20/75). Changes in pH, color, thiobarbituric acid reactive substances (TBARS), Warner−Bratzler shear force (WBSF), and microbiology (total viable counts, Enteriobacteriaceae, and lactic acid bacteria) were monitored over 15 days post-mortem at 4 °C. VIS-NIR spectra were collected from pork fillets before (through the film cover) and after opening the trays (directly on the meat surface) with a portable LABSPEC 5000 NIR system in diffuse reflectance mode (350−2500 nm). Quantitative NIR models by partial least squares regression (PLSR) showed a promising prediction ability for meat color (L*, a*, C*, and h*) and microbiological variables (R2VAL > 0.72 and RPDVAL > 2). In addition, qualitative models using PLS discriminant analysis obtained good accuracy (over 90%) for classifying pork samples as fresh (acceptable for consumption) or spoiled (not acceptable) based on their microbiological counts. VIS-NIR spectroscopy allows rapid evaluation of product quality and shelf life and could be used for on-site control of pork quality.
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The potential of near-infrared reflectance spectroscopy (NIRS) to discriminate Normal and DFD (dark, firm, and dry) beef and predict quality traits in 129 Longissimus thoracis (LT) samples from three Spanish purebreeds, Asturiana de los Valles (AV; n = 50), Rubia Gallega (RG; n = 37), and Retinta (RE; n = 42) was assessed. The results obtained by partial least squares-discriminant analysis (PLS-DA) indicated successful discrimination between Normal and DFD samples of meat from AV and RG (with sensitivity over 93% for both and specificity of 100 and 72%, respectively), while RE and total sample sets showed poorer results. Soft independent modelling of class analogies (SIMCA) showed 100% sensitivity for DFD meat in total, AV, RG, and RE sample sets and over 90% specificity for AV, RG, and RE, while it was very low for the total sample set (19.8%). NIRS quantitative models by partial least squares regression (PLSR) allowed reliable prediction of color parameters (CIE L*, a*, b*, hue, chroma). Results from qualitative and quantitative assays are interesting in terms of early decision making in the meat production chain to avoid economic losses and food waste.
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This study investigated the effect of different cattle management strategies at farm (Intensive vs. Extensive) and during transport and lairage (mixing vs. non-mixing with unfamiliar animals) on the myofibrillar subproteome of Longissimus thoracis et lumborum (LTL) muscle of "Asturiana de los Valles" yearling bulls. It further aimed to study the relationships with beef quality traits including pH, color, and tenderness evaluated by Warner-Bratzler shear force (WBSF). Thus, comparative proteomics of the myofibrillar fraction along meat maturation (from 2 h to 14 days post-mortem) and different quality traits were analyzed. A total of 23 protein fragments corresponding to 21 unique proteins showed significant differences among the treatments (p < 0.05) due to any of the factors considered (Farm, Transport and Lairage, and post-mortem time ageing). The proteins belong to several biological pathways including three structural proteins (MYBPC2, TNNT3, and MYL1) and one metabolic enzyme (ALDOA) that were affected by both Farm and Transport/Lairage factors. ACTA1, LDB3, and FHL2 were affected by Farm factors, while TNNI2 and MYLPF (structural proteins), PKM (metabolic enzyme), and HSPB1 (small Heat shock protein) were affected by Transport/Lairage factors. Several correlations were found between the changing proteins (PKM, ALDOA, TNNI2, TNNT3, ACTA1, MYL1, and CRYAB) and color and tenderness beef quality traits, indicating their importance in the determination of meat quality and their possible use as putative biomarkers.
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The objective of this work was to demonstrate how the extraction method affects the reliability of biomarker detection and how this detection depends on the biomarker location within the cell compartment. Different extraction methods were used to study the sarcoplasmic and myofibrillar fractions of the Longissimus thoracis et lumborum muscle of young bulls of the Asturiana de los Valles breed in two quality grades, standard (Control) or dark, firm, and dry (DFD) meat. Protein extractability and the expression of some of the main meat quality biomarkers-oxidative status (lipoperoxidation (LPO) and catalase activity (CAT)), proteome (SDS-PAGE electrophoretic pattern), and cell stress protein (Hsp70)-were analyzed. In the sarcoplasmic fraction, buffers containing Triton X-100 showed significantly higher protein extractability, LPO, and higher intensity of high-molecular-weight protein bands, whereas the TES buffer was more sensitive to distinguishing differences in the protein pattern between the Control and DFD meat. In the myofibrillar fraction, samples extracted with the lysis buffer showed significantly higher protein extractability, whereas samples extracted with the non-denaturing buffer showed higher results for LPO, CAT, and Hsp70, and higher-intensity bands in the electrophoretic pattern. These findings highlight the need for the careful selection of the extraction method used to analyze the different biomarkers considering their cellular location to adapt the extractive process.
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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.
Assuntos
Animais Domésticos/genética , Músculo Esquelético/metabolismo , Proteômica , Biologia de Sistemas , Animais , Autofagia , CarneRESUMO
The conversion of muscle into meat is a complex process of major concern for meat scientists due to its influence on the final meat quality. The aim of this study was to investigate the occurrence of autophagic processes in the conversion of muscle into meat. Our findings demonstrated, for the first time, the occurrence of autophagic processes in the muscle tissue at early postmortem period (2 h to 24 h) in both beef breeds studied (Asturiana de los Valles and Asturiana de la Montaña) showing significant time-scale differences between breeds, which could indicate a role of this process in meat maturation. These breeds have different physiological features: while Asturiana de los Valles is a meat-specialized breed showing high growth rate, an elevated proportion of white fibers in the muscle and low intramuscular fat level, Asturiana de la Montaña is a small- to medium-sized rustic breed adapted to less-favored areas, showing more red fibers in the muscle and a high intramuscular fat content.
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Autofagia , Carne , Animais , Antioxidantes/metabolismo , Cruzamento , Catepsinas/metabolismo , Bovinos , Immunoblotting , Proteínas de Membrana/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Fatores de TempoRESUMO
The Syrian hamster Harderian gland (HG) has a marked sexual dimorphism and exhibits an extraordinary rate of porphyrinogenesis. The physiological oxidative stress, derived from constant porphyrin production, is so high that the HG needs additional survival autophagic mechanisms to fight against this chronic exposure, provoking the triggering of a holocrine secretion in female glands that forms two types of secretory masses: intra-tubular-syncytial and inter-tubular-syncytial masses. The aim of this work was to study the development of this inter-tubular holocrine secretion. To approach this task, we have considered that the steps developed during the formation of the so-called invasive masses consist of the growth of epithelial cells, cell detachment from the basal lamina and invasion of surrounding tissues. The presence of these masses, particularly in the female HG, are closely linked to sexual dimorphism in redox balance and to alterations in the expression of certain factors such as cytokeratins, P-cadherin, matrix metalloproteinases, cathepsin H, proliferating cell nuclear antigen, p53, CD-31 and vascular endothelial growth factor, which seem to be involved in tissue remodeling. The results document unusual mechanisms of secretion in Syrian hamster HG: an extraordinary system of massive secretion through the conjunctive tissue, disrupting the branched structure of the gland.
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Glândula de Harder/anatomia & histologia , Mesocricetus/anatomia & histologia , Animais , Autofagia/fisiologia , Caderinas/metabolismo , Catepsina H/metabolismo , Cricetinae , Feminino , Glândula de Harder/fisiologia , Glândula de Harder/ultraestrutura , Imuno-Histoquímica , Queratinas/metabolismo , Peroxidação de Lipídeos/fisiologia , Masculino , Mesocricetus/fisiologia , Estresse Oxidativo/fisiologia , Caracteres SexuaisRESUMO
The possibility that mitochondria are involved in cellular dysfunction is particularly high in situations associated with increases in free radical activity, like hypoxia or ischemia; therefore its potential role in the muscle post-mortem metabolism is reviewed. In the dying muscle, different routes of cell death catabolism (apoptosis, autophagy) may occur having great influence on the process of conversion of muscle into meat. Mitochondria are the first and also one of the main organelles affected by post-mortem changes; therefore they are decisive in the subsequent cellular responses influencing the pathway to cell demise and thus, the final meat quality. Depending on the cell death programme followed by muscle cells after exsanguination, diverse proteases would be activated to a different extent, which is also reviewed in order to understand how they affect meat tenderization. This review also summarizes recent patents relating cell death processes and meat tenderness. Further research is encouraged as there is still a need of knowledge on cell death post-mortem processes to increase our understanding of the conversion of muscle into meat.
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Apoptose , Autofagia , Tecnologia de Alimentos , Carne , Mitocôndrias Musculares/metabolismo , Músculo Esquelético/metabolismo , Matadouros , Animais , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Manipulação de Alimentos , Qualidade dos Alimentos , Humanos , Mitocôndrias Musculares/efeitos dos fármacos , Mitocôndrias Musculares/patologia , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/patologia , Mudanças Depois da MorteRESUMO
Se describe el caso de una mujer en su primer embarazo, con retención aguda urinaria en asociación con útero grávido en retroversión. En su segundo embarazo, el proceso de incarceración de un útero grávido sucedió de nuevo. Ha sido tratada, en ambos casos con éxito, con la reposición manual del útero grávido. En el tercer embarazo, en la décima semana, se coloca un pesario en la vagina, evitando la incarceración del útero. Se discuten varios aspectos de la frecuencia, la etiología, el tratamiento y la posibilidad de recidiva del útero incarcerado en retroversión (AU)
We describe the case of a woman in her first pregnancy with acute urinary retention associated with a retroverted gravid uterus. In her second pregnancy, the process of incarceration of a gravid uterus recurred. On both occasions, the patient was successfully treated by manual repositioning of the gravid uterus. In the tenth week of the patient's third pregnancy, a pessary was placed in the vagina, which prevented uterine incarceration. Several aspects of the frequency, etiology, treatment and possibility of recurrent incarcerated retroverted uterus are discussed (AU)
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Humanos , Feminino , Gravidez , Adulto , Retroversão Uterina/complicações , Retroversão Uterina/diagnóstico , Retenção Urinária/complicações , Retenção Urinária/diagnóstico , Complicações na Gravidez/epidemiologia , Retroversão Uterina/fisiopatologia , Retroversão Uterina , Diagnóstico DiferencialRESUMO
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.
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Calpaína/metabolismo , Carne/análise , Músculo Esquelético/metabolismo , Mudanças Depois da Morte , Animais , Biomarcadores , Bovinos , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Concentração de Íons de Hidrogênio , Hipertrofia , Carne/classificação , Músculo Esquelético/enzimologia , Miofibrilas/metabolismo , Peptídeos/metabolismo , Proteólise , Retículo Sarcoplasmático/metabolismo , Especificidade da Espécie , Troponina T/metabolismoRESUMO
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.
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Antioxidantes/farmacologia , Autofagia/efeitos dos fármacos , Glândula de Harder/efeitos dos fármacos , Glândula de Harder/metabolismo , Melatonina/farmacologia , Animais , Caspase 3/metabolismo , Catalase/metabolismo , Catepsina B/metabolismo , Cricetinae , Feminino , Glândula de Harder/química , Glândula de Harder/citologia , Peroxidação de Lipídeos/efeitos dos fármacos , Mesocricetus , NF-kappa B/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Carbonilação Proteica/efeitos dos fármacos , Superóxido Dismutase/metabolismo , Proteína Supressora de Tumor p53/metabolismoRESUMO
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.
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Envelhecimento/imunologia , Autofagia/imunologia , NF-kappa B/fisiologia , Estresse Oxidativo/imunologia , Animais , Antioxidantes/administração & dosagem , Apoptose , Autofagia/efeitos dos fármacos , Catalase/metabolismo , Catepsina B/metabolismo , Catepsina D/metabolismo , Feminino , Masculino , Melatonina/administração & dosagem , Camundongos , NF-kappa B/análise , Antígeno Nuclear de Célula em Proliferação/análise , Receptores de Melatonina/análise , Fatores Sexuais , Transdução de Sinais/efeitos dos fármacos , Baço/enzimologia , Baço/imunologiaRESUMO
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.
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Autofagia/fisiologia , Glândula de Harder , Caracteres Sexuais , Animais , Catalase/metabolismo , Catepsinas/metabolismo , Morte Celular/fisiologia , Cricetinae , Cricetulus , Feminino , Glândula de Harder/anatomia & histologia , Glândula de Harder/metabolismo , Glândula de Harder/fisiologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Fígado/citologia , Fígado/metabolismo , Masculino , Mesocricetus , NF-kappa B/metabolismo , Estresse Oxidativo , Porfirinas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sirtuínas/metabolismo , Superóxido Dismutase/metabolismo , Proteína Supressora de Tumor p53/metabolismoRESUMO
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.
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Androgênios/metabolismo , Autofagia/fisiologia , Glândulas Sebáceas/metabolismo , Glândulas Sebáceas/ultraestrutura , Caracteres Sexuais , Animais , Apoptose/fisiologia , Proteínas Reguladoras de Apoptose/metabolismo , Western Blotting , Caspase 3/metabolismo , Catepsina D/metabolismo , Cricetinae , Feminino , Queratinas/metabolismo , Masculino , Mesocricetus , Microscopia Eletrônica de TransmissãoRESUMO
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.