Plastid transformation: advances and challenges for its implementation in agricultural crops

Chloroplast biotechnology has emerged as a promissory platform for the development of modified plants to express products aimed mainly at the pharmaceutical, agricultural, and energy industries. This technology's high value is due to its high capacity for the mass production of proteins. Moreover, the interest in chloroplasts has increased because of the possibility of expressing multiple genes in a single transformation event without the risk of epigenetic effects. Although this technology solves several problems caused by nuclear genetic engineering, such as turning plants into safe bio-factories, some issues must still be addressed in relation to the optimization of regulatory regions for efficient gene expression, cereal transformation, gene expression in non-green tissues, and low transformation efficiency. In this article, we provide information on the transformation of plastids and discuss the most recent achievements in chloroplast bioengineering and its impact on the biopharmaceutical and agricultural industries; we also discuss new tools that can be used to solve current challenges for their successful establishment in recalcitrant crops such as monocots.

Lycopene and Melatonin: Antioxidant Compounds in Cosmetic Formulations.

BACKGROUND: The use of antioxidants has become a common practice in the development of antiaging cosmetics. OBJECTIVE: The aim of this study was to evaluate the clinical efficacy of cosmetic formulations containing lycopene and melatonin antioxidants. METHOD: Thirty-six healthy women from 32 to 65 years were enrolled in this study. The study was carried out for 10 weeks, 2 preconditioning weeks with a control cream without antioxidants, and 8-week test with creams containing antioxidants in study. A multifunctional skin physiology monitor (Courage & Khazaka electronic GmbH®, Germany) was used to measure skin sebum content, hydration, elasticity, erythema index, and melanin index in 4 different regions of the face. RESULTS: There were significant differences between them.

Factors that bias teacher expectations: Findings from Chile / Factores que sesgan las expectativas docentes: hallazgos en Chile

Abstract It is well-known that teacher expectations tend to be biased by factors such as student socio-economic status (SES) and gender. However, much less research has been devoted to understanding how teacher characteristics may impact their own expectations of the students. The present study investigated teacher expectations for 343 Chilean teachers (240 in-service and 103 pre-service). We first designed and validated an instrument to measure expectations; then we assessed the effect of teacher gender and experience, and student gender and school-SES in the formation of teacher expectations. The data were analysed using hypothesis and data-driven analyses. The results showed that SES had an effect on teacher expectations (η2= .03 to .12); there was a higher probability that teachers from high-SES schools would have positive expectations of their students. However, negative expectations were equally distributed across teachers working in high and low-SES schools. There was also no evidence of the effect of teacher or student gender on teacher expectations. With respect to teacher experience, the findings were clear cut; both pre-service and in-service teachers shared identical expectations of their students. These findings have important implications regarding teacher training programmes since the expectation bias is observed very early during training.

Resumen Es bien sabido que las expectativas docentes tienden a estar sesgadas por factores como el nivel socioeconómico de los estudiantes (NSE) y el género. Sin embargo, se ha investigado menos cómo las características de los docentes pueden afectar sus propias expectativas de los estudiantes. El presente estudio investigó las expectativas de 343 docentes chilenos (240 en servicio y 103 alumnos en prácticas). Primero se diseñó y validó un instrumento para medir expectativas; luego se evaluó el efecto del género y experiencia del profesor, y el género del estudiante y NSE de la escuela en la formación de las expectativas del profesor. Los datos fueron analizados mediante técnicas dirigidas por hipótesis y por datos. Los resultados mostraron un efecto del NSE de la escuela en las expectativas docentes (η2 = .03 a .12), con una mayor probabilidad de que los profesores de escuelas con alto NSE tengan expectativas positivas de sus estudiantes. Sin embargo, las expectativas negativas fueron distribuidas equitativamente entre los profesores que trabajaban en escuelas con bajo y alto NSE. Tampoco hubo evidencia del efecto de género de los profesores o estudiantes en las expectativas del profesor. Respecto a la experiencia del profesor, los hallazgos son claros; tanto los estudiantes en práctica profesional como los profesores en servicio mostraron idénticas expectativas hacia sus estudiantes. Estos hallazgos tienen implicaciones importantes respecto a los programas de formación docente ya que el sesgo de las expectativas se observa tempranamente desde la formación.

Body weight gain in rats by a high-fat diet produces chronodisruption in activity/inactivity circadian rhythm.

In the last few decades, obesity has become one of the most important public health problems. Adipose tissue is an active endocrine tissue which follows a rhythmic pattern in its functions and may produce alterations in certain circadian rhythms. Our aim was to evaluate whether the locomotor activity circadian rhythm could be modified by a hypercaloric diet in rodents. Two groups were considered in the experiment: 16 rats were used as a control group and were fed standard chow; the other group comprised 16 rats fed a high-fat diet (35.8% fat, 35% glucides). The trial lasted 16 weeks. Body weight was measured every week, and a blood sample was extracted every two weeks to quantify triglyceride levels. The activity/inactivity circadian rhythm was logged through actimetry throughout the trial, and analysed using the DAS 24© software package. At the end of the experiment, the high-fat fed rats had obese-like body weights and high plasma triglyceride levels, and, compared with the control group, increased diurnal activity, decreased nocturnal activity, reductions in amplitude, midline estimating statistic of rhythm, acrophase and interdaily stability, and increases in intradaily variability of their activity rhythms. The results thus show how obesity can lead to symptoms of chronodisruption in the body similar to those of ageing.

A lycopene-enriched virgin olive oil enhances antioxidant status in humans.

BACKGROUND: Lycopene, a bioactive red pigment, represents the most potent in vitro antioxidant among carotenoids. Virgin olive oil contains trace amounts of a wide variety of phytochemicals, which have proven to exert beneficial effects on oxidative stress. Since the ingestion of lycopene together with oil reportedly increases its bioavailability, we evaluated urinary antioxidant capacity after the consumption of a lycopene-enriched virgin olive oil (7 mg lycopene day(-1)) compared with the antioxidant effect produced after the ingestion of a virgin olive oil and a sunflower oil during 5 days, in young (25-30 years of age), middle-aged (35-55 years of age) and elderly (65-85 years of age) subjects. RESULTS: The results showed that the consumption of virgin olive oil increased urinary antioxidant capacity in middle-aged and elderly volunteers, whereas the administration of a lycopene-enriched virgin olive oil produced higher antioxidant effects in all of the three age groups assayed. CONCLUSION: The incorporation of the lycopene-enriched virgin olive oil into the diet may enhance the health-promoting effects of the virgin olive oil, contributing as a functional tool against several disorders where oxidative stress plays an important role.

Systemic inflammatory load in young and old ringdoves is modulated by consumption of a Jerte Valley cherry-based product.

A chronic subclinical inflammatory status that coexists with immune dysfunction is commonly found in the elderly population. Consumption of foods rich in antioxidants (e.g., cherries) is an attractive strategy to reduce risk from chronic diseases. Based on previous studies showing the antioxidant effect of a Jerte Valley cherry derivative product in humans, the objective of this work was to evaluate the effect of the intake of a Jerte Valley cherry-based beverage on inflammatory load in both young and old ringdoves (Streptopelia risoria). To this purpose, circulating levels of pro-inflammatory and anti-inflammatory cytokines as well as serum levels of different acute-phase proteins were measured before and after a 10-day treatment with the Jerte Valley cherry-based beverage. Thus, the 10-day treatment with the cherry-based beverage modulated the balance of pro- and anti-inflammatory cytokines in both young and old ringdoves by down-regulating the levels of pro-inflammatory cytokines (interleukin [IL]-1ß, tumor necrosis factor-α, and interferon-γ) and up-regulating the levels of anti-inflammatory cytokines (IL-4, IL-2, and IL-10). Moreover, the 10-day treatment with the Jerte Valley cherry-based product reduced the levels of several proteins involved in acute-phase responses, such as C-reactive protein, haptoglobin, α(2)-macroglobulin, and serum amyloid P component. On the other hand, old birds showed imbalanced levels of inflammatory markers toward a pro-inflammatory status, thereby underlining the fact that aging is usually accompanied by systemic inflammation and inflammation-related chronic diseases. To sum up, the data suggest a potential health benefit by consuming the cherry-based beverage, especially in aged populations, through their anti-inflammatory properties.

The sedative effect of non-alcoholic beer in healthy female nurses.

INTRODUCTION: The hop (Humulus lupulus L.), a component of beer, is a sedative plant whose pharmacological activity is principally due to its bitter resins, in particular to the α-acid degradation product 2-methyl-3-buten-2-ol. The mechanism of action of hop resin consists of raising the levels of the neurotransmitter γ-aminobutyric acid (GABA), an inhibitory neurotransmitter acting in the central nervous system (CNS). OBJECTIVES: To analyze the sedative effect of hops as a component of non-alcoholic beer on the sleep/wake rhythm in a work-stressed population. METHODS: The experiment was conducted with healthy female nurses (n = 17) working rotating and/or night shifts. Overnight sleep and chronobiological parameters were assessed by actigraphy (Actiwatch®) after moderate ingestion of non-alcoholic beer containing hops (333 ml with 0.0% alcohol) with supper for 14 days (treatment). Data were obtained in comparison with her own control group without consumption of beer during supper. RESULTS: Actigraphy results demonstrated improvement of night sleep quality as regards the most important parameters: Sleep Latency diminished (p≤0.05) in the Treatment group (12.01±1.19 min) when compared to the Control group (20.50±4.21 min), as also did Total Activity (p≤0.05; Treatment group = 5284.78±836.99 activity pulses vs Control = 7258.78±898.89 activity pulses). In addition, anxiety as indexed by the State-Trait Anxiety Inventory (STAI) decreased in the Treatment group (State Anxiety 18.09±3.8 vs Control 20.69±2.14). CONCLUSION: The moderate consumption of non-alcoholic beer will favour night-time rest, due in particular to its hop components, in addition to its other confirmed benefits for the organism.

The consumption of a Jerte Valley cherry product in humans enhances mood, and increases 5-hydroxyindoleacetic acid but reduces cortisol levels in urine.

PURPOSE: Jerte Valley cherries contain high levels of tryptophan, serotonin, and melatonin. These molecules have been shown to be involved in mood regulation. It has been suggested that a complex inter-relationship between brain serotonin, circulating levels of cortisol (the major stress hormone), and the hypothalamus-pituitary-adrenal axis exists in the regulation of stress responses, where cortisol and serotonin act as markers of mood disturbances. Moreover there is growing evidence that altered HPA activity is associated with various age-related pathologies. The present study evaluated the effect of the ingestion of a Jerte Valley cherry-based product, compared to a placebo product, on urine cortisol and 5-hydroxyindoleacetic acid (5-HIAA) levels, and on mood in young, middle-aged, and elderly participants. METHODS: Cortisol and 5-HIAA acid levels were measured by commercial enzyme-linked immunosorbent assay kits. The mood state profile was analysed using a visual analogue scale and the state-trait anxiety inventory. RESULTS: Our findings showed that the ingestion of the Jerte Valley cherry product decreased urinary cortisol and increased urinary 5-HIAA levels in all the experimental groups. Moreover, the cherry product was able to lessen anxiety status in the middle-aged and elderly participants, and enhanced subjective mood parameters, particularly family relationships in young participants, and frame of mind and fitness in both middle-aged and elderly subjects. CONCLUSIONS: The consumption of the Jerte Valley cherry product may protect against stress and act as a mood enhancer by increasing serotonin availability to the organism, particularly with advancing age.

A jerte valley cherry-based product as a supply of tryptophan.

L-Tryptophan (tryptophan) is an essential amino acid in humans. It has important roles as a precursor of different bioactive compounds. Based on previous studies in which tryptophan has been shown to be present in fresh cherries, the aim of the present work was to analyze the tryptophan content of a Jerte Valley cherry-based product. A previously optimized method of analysis of tryptophan was used, ie, high-performance liquid chromatography with fluorescence detection (HPLC/FL). As expected, HPLC/FL technique permitted to detect and quantify the tryptophan content in a different matrix rather than fresh cherries. In fact, the Jerte Valley cherry-based product contained 69.54 ± 10.64 ppm of tryptophan, thereby showing that this product is a good source of tryptophan. In summary, it has been proven that the Jerte Valley cherry-based product is rich in tryptophan and may be indicated as a supply of this essential amino acid as well as having potential health benefits for conditions where tryptophan is necessary.

Urinary 6-sulfatoxymelatonin and total antioxidant capacity increase after the intake of a grape juice cv. Tempranillo stabilized with HHP.

Red grapes contain elevated amounts of antioxidant compounds (polyphenols) that may potentially prevent cell aging, cardiovascular disease and oxidation-related disorders. Since functional drinks are presently one of the most dynamic sectors of the market, the present work was aimed at evaluating the possible antioxidant effect of an experimental grape juice in terms of urinary 6-sulfatoxymelatonin (aMT6-s) and total antioxidant capacity in young (20 ± 10 yr-old), middle-aged (45 ± 10 yr-old) and elderly (75 ± 10 yr-old) individuals. Grapes (Vitis vinifera cv. Tempranillo) were de-stemmed, racked and pressed. The juice was subsequently stabilized by high hydrostatic pressure (HHP). Participants consumed 200 mL of grape juice twice a day (as the lunch and dinner desserts) for 5 days. First-void morning urines were collected before treatment (basal values), the day immediately after the last ingestion of juice (assay), and one day afterwards (post-assay). aMT6-s and total antioxidant capacity were quantified using commercial ELISA and colorimetric assay kits, respectively. The intake of grape juice cv. Tempranillo induced a significant increase of urinary aMT6-s and total antioxidant capacity in the three groups of age analyzed as compared to their corresponding basal and post-assay values. These functional/nutraceutical properties may be of interest for a prospective commercialization of the grape juice. The novel technology used for juice stabilization may be suitable for introducing into the market a product with high sensory and nutritional quality, as it has been shown in this study.

Protective effect of melatonin against human leukocyte apoptosis induced by intracellular calcium overload: relation with its antioxidant actions.

Apoptosis or programmed cell death plays a critical role in both inflammatory and immune responses. Recent evidence demonstrates that control of leukocyte apoptosis is one of the most striking immune system-related roles of melatonin. For this reason, this study evaluated the protective effects of melatonin on human leukocyte apoptosis induced by sustained cytosolic calcium increases. Such protective effects are likely mediated by melatonin's free-radical scavenging actions. Treatments with the specific inhibitor of cytosolic calcium re-uptake, thapsigargin (TG), and/or the calcium-mobilizing agonist, N-formyl-methionyl-leucyl-phenylalanine (FMLP), induced intracellular reactive oxygen species (ROS) production, caspase activation as well as DNA fragmentation in human leukocytes. Also, TG- and/or FMLP-induced apoptosis was dependent on both cytosolic calcium increases and calcium uptake into mitochondria, because when cells were preincubated with the cytosolic calcium chelator, dimethyl BAPTA, and the inhibitor of mitochondrial calcium uptake, Ru360, TG- and FMLP-induced apoptosis was largely inhibited. Importantly, melatonin treatment substantially prevented intracellular ROS production, reversed caspase activation, and forestalled DNA fragmentation induced by TG and FMLP. Similar results were obtained by preincubating the cells with another well-known antioxidant, i.e., N-acetyl-L-cysteine. To sum up, depletion of intracellular calcium stores induced by TG and/or FMLP triggers different apoptotic events in human leukocytes that are dependent on calcium signaling. The protective effects resulting from melatonin administration on leukocyte apoptosis likely depend on melatonin's antioxidant action because we proved that this protection is melatonin receptor independent. These findings help to understand how melatonin controls apoptosis in cells of immune/inflammatory relevance.

Melatonin enhances hydrogen peroxide-induced apoptosis in human promyelocytic leukaemia HL-60 cells.

Melatonin is an indoleamine secreted by the pineal gland that shows multiple tasks. This ubiquitously acting free radical scavenger has recently been shown to stimulate the production of reactive oxygen species (ROS) in tumour cells, making them undergo apoptosis, whilst it prevents apoptosis in healthy cells. The mechanisms by which melatonin exerts these dual actions are, however, not yet clearly understood. Thus, the aim of this study was to further investigate how melatonin can enhance oxidative stress-induced apoptosis in a leukaemia cell line. The results show that melatonin increased the apoptotic effects of H(2)O(2) in human myeloid HL-60 cells as assessed by cellular viability, mitochondrial permeability transition induction, mitochondrial membrane depolarization, ROS generation, caspases 3, 8 and 9 activity, phosphatidylserine externalization, and DNA fragmentation techniques. When healthy leucocytes were exposed to H(2)O(2), melatonin increased the viability of the cells. Taken together, the findings indicate that melatonin is a potential physiological tool capable of protecting healthy cells from chemotherapy-induced ROS production as well as inducing tumour cell death. Because cancer cells manifest increased oxidative stress as a result of their elevated metabolism, the use of melatonin may be useful in impairing their ROS buffering capacity.

Melatonin is able to delay endoplasmic reticulum stress-induced apoptosis in leukocytes from elderly humans.

The mechanisms regulating neutrophil apoptosis are basically unaffected by the aging process. However, a significant impairment of cell survival occurs in elderly individuals following neutrophil challenge with pro-inflammatory stimuli, such as granulocyte-macrophage colony-stimulating factor (GM-CSF). The goal of the present study was to prove the effects of melatonin supplementation on apoptosis induced by calcium signaling in human leukocytes from elderly volunteers. Treatments with the specific inhibitor of cytosolic calcium re-uptake, thapsigargin, and/or the calcium mobilizing agonist, N-formyl-methionyl-leucyl-phenylalanine (fMLP), induced mitochondrial membrane depolarization, caspase activation, phosphatidylserine (PS) externalization, and DNA fragmentation in leukocytes from both young and elderly volunteers, although such effects were much more evident in aged leukocytes. Importantly, melatonin treatment substantially preserved mitochondrial membrane potential, reversed caspase activation, reduced PS exposure and forestalled DNA fragmentation in leukocytes from both age groups. In conclusion, melatonin is able to delay endoplasmic reticulum stress-induced apoptosis in aged leukocytes and may counteract, at the cellular level, age-related degenerative phenomena linked to oxidative stress.

Pro-oxidant effect of melatonin in tumour leucocytes: relation with its cytotoxic and pro-apoptotic effects.

Melatonin has many effects on a wide range of physiological functions and is involved in a number of pathological events including oncostatic and neoplastic processes. The tissue protective actions of melatonin are attributed to its well-known antioxidant activity though melatonin might also exert pro-oxidant effects, particularly in tumour cells. This study evaluated the pro-oxidant effects of melatonin in tumour cell lines of human haematopoietic origin. Melatonin treatment is able to stimulate production of intracellular reactive oxygen species (ROS), as revealed by the increase in rhodamine-123 fluorescence, which was associated with significant cytotoxicity and activation of caspase activities. Furthermore, pre-treatment of cells with well-known antioxidants, such as N-acetyl-L-cysteine (NAC), trolox, PEG-catalase and reduced glutathione (GSH), reversed the effects of melatonin on both intracellular ROS production, as on the cytotoxicity and caspase activation. This pro-oxidant action of melatonin may assist in limiting tumour cell growth.

Jerte Valley cherry-enriched diets improve nocturnal rest and increase 6-sulfatoxymelatonin and total antioxidant capacity in the urine of middle-aged and elderly humans.

Tryptophan, serotonin, and melatonin, present in Jerte Valley cherries, participate in sleep regulation and exhibit antioxidant properties. The effect of the intake of seven different Jerte Valley cherry cultivars on the sleep-wake cycle, 6-sulfatoxymelatonin levels, and urinary total antioxidant capacity in middle-aged and elderly participants was evaluated. Volunteers were subjected to actigraphic monitoring to record and display the temporal patterns of their nocturnal activity and rest. 6-sulfatoxymelatonin and total antioxidant capacity were quantified by enzyme-linked immunosorbent assay and colorimetric assay kits, respectively. The intake of each of the cherry cultivars produced beneficial effects on actual sleep time, total nocturnal activity, assumed sleep, and immobility. Also, there were significant increases in 6-sulfatoxymelatonin levels and total antioxidant capacity in urine after the intake of each cultivar. These findings suggested that the intake of Jerte Valley cherries exerted positive effect on sleep and may be seen as a potential nutraceutical tool to counteract oxidation.

Melatonin counteracts alterations in oxidative metabolism and cell viability induced by intracellular calcium overload in human leucocytes: changes with age.

Ageing is associated with an increased production of free radicals and alterations in the mechanisms of adaptation to oxidative stress. In fact, the free radical theory of ageing proposes that deleterious actions of free radicals are responsible for the functional deterioration associated with ageing. Moreover, a close relationship exists between calcium homeostasis and oxidative stress. The current work was aimed at proving that intracellular calcium overload induced by N-formyl-methionyl-leucyl-phenylalanine (FMLP) and/or thapsigargin leads to oxidative stress. We additionally examined the effect of melatonin on the levels of reactive oxygen species (ROS) and cell viability in human leucocytes collected from young (20-30-year-old) and elderly (65-75-year-old) individuals under both basal and oxidative stress-induced conditions. Treatments with 10 nM FMLP and/or 1 microM thapsigargin induced a transient increase in cytosolic free-calcium concentration ([Ca(2 + )](c)) in human leucocytes due to calcium release from internal stores, and led in turn to oxidative stress, as assessed by intracellular ROS measurement. Non-treated leucocytes from aged individuals exhibited higher ROS levels and lower rates of cell survival when compared to leucocytes from young individuals. Similar results were obtained in FMLP and/or thapsigargin-treated leucocytes from elderly individuals when compared to those from the young individuals. Melatonin treatment significantly reduced both hydrogen peroxide (H(2)O(2)) and superoxide anion levels, likely due to its free-radical scavenging properties, and enhanced leucocyte viability in both age groups. Therefore, melatonin may be a useful tool for the treatment of disease states and processes where an excessive production of oxidative damage occurs.

Melatonin reduces apoptosis induced by calcium signaling in human leukocytes: Evidence for the involvement of mitochondria and Bax activation.

We have evaluated the effect of melatonin on apoptosis evoked by increases in [Ca(2+)]( c ) in human leukocytes. Our results show that treatment of neutrophils with the calcium mobilizing agonist FMLP or the specific inhibitor of calcium reuptake thapsigargin induced a transient increase in [Ca(2+)]( c ). Our results also show that FMLP and thapsigargin increased caspase-9 and -3 activities and the active forms of both caspases. The effect of FMLP and thapsigargin on caspase activation was time-dependent. Similar results were obtained when lymphocytes were stimulated with thapsigargin. This stimulatory effect was accompanied by induction of mPTP, activation of the proapoptotic protein Bax and release of cytochrome c. However, when leukocytes were pretreated with melatonin, all of the apoptotic features indicated above were significantly reversed. Our results suggest that melatonin reduces caspase-9 and -3 activities induced by increases in [Ca(2+)]( c ) in human leukocytes, which are produced through the inhibition of both mPTP and Bax activation.

Improving the quality of infant sleep through the inclusion at supper of cereals enriched with tryptophan, adenosine-5'-phosphate, and uridine-5'-phosphate.

The present study evaluated whether the administration of cereals enriched with nutrients that are facilitators of sleep could help improve the sleep of infants who had sleep disorders at night time. Thirty infants aged 8-16 months with sleep disorders involving at least three nocturnal waking episodes took part in the study. They were given a night-time 'sleep facilitating cereal' product containing 225 mg tryptophan, 5.3 mg adenosine-5'-P, and 6.3 mg uridine-5'-P per 100 g of product. These cereals were given in a double-blind procedure lasting 5 weeks, with ingestion of the cereal between 18:00 and 06:00. In the control week, the children received a standard cereal (75 mg tryptophan/100 g product without nucleotides) dissolved in a standard formula milk (231.5 mg tryptophan, 2.6 mg adenosine-5'-P, 5 mg uridine-5'-P, per 100 g product). In one experimental week, the children received the night-time sleep facilitating cereal together with the standard formula milk. In another week, they received the sleep facilitating cereal together with a night milk specially formulated to attain the sleep rhythm (480 mg tryptophan, 8.8 mg uridine-5'-P, and 7.6 mg adenosine-5'-P per 100 g product). The three experimental weeks were separated by two wash-out weeks in which the milk and cereal administered was identical in composition to that of the control week. All the infants received a programmed writer actimeter which they wore continually, attached to their ankles, to record their motor activity. The recorded activity was used to calculate information about the time in bed, assumed sleep, actual sleep, sleep efficiency, sleep latency, immobility, and total activity. The infants receiving the enriched cereal during the time of darkness showed improvements in their sleep parameters, regardless of whether the milk they took at night was standard or enriched with tryptophan, adenosine-5'-P, and uridine-5'-P. In summary, the administration of enriched cereals led to an improvement in sleep, regardless of the type of infant milk used. These results support the concept of chrononutrition since they confirm that the sleep/wake rhythm can be influenced by diet.

Melatonin and tryptophan counteract lipid peroxidation and modulate superoxide dismutase activity in ringdove heterophils in vivo. Effect of antigen-induced activation and age.

Aging is associated with an increased production of free radicals and alterations in the mechanisms of adaptation to stress. A number of studies have shown a causal connection between age-related oxidant/antioxidant imbalance and the diminution of an organism's melatonin levels in old age. Restoration of this rhythm may contribute to the re-stabilization of cellular homeostasis. The present work was aimed at examining the effect of the administration of melatonin or its precursor, the amino acid tryptophan, on heterophil lipid peroxidation (LPO) levels and superoxide dismutase (SOD) activity in heterophils from young (4-5 year) and old (12-14 year) ringdoves (Streptopelia risoria) under both basal and antigen-induced (stressful) conditions. Young animals were treated for 3 consecutive days with a single daily oral dose (0.25 mg) of melatonin at 1900 hours while old animals were treated for 3 consecutive days with either an oral dose (2.5 mg) of melatonin at 1900 hours or an oral capsule of 300 mg/kg body weight of L-tryptophan at 0900 hours. Non-treated old animals presented lower circulating levels of melatonin at night and higher levels of malonaldehyde (MDA) both day and night when compared to young birds. In both age groups, LPO levels were lower at night than during the day. Melatonin or tryptophan treatments significantly increased serum melatonin levels, reinforced SOD activity, and reduced MDA levels induced by the antigen. Melatonin and tryptophan may be useful agents for the treatment of disease states and processes in which an excessive production of oxidative damage occurs.

Melatonin induces mitochondrial-mediated apoptosis in human myeloid HL-60 cells.

The role of melatonin in the mediation of apoptotic events has recently gained attention, especially after recent studies have reported that melatonin exerts antiapoptotic actions in normal cells but may activate proapoptotic pathways in some tumor cells. Here, we have evaluated the effect of melatonin on apoptosis in the human leukemia cell line HL-60. Melatonin treatment (1 mm) induced a significant increase in caspase-3 and -9 activities. The effect of melatonin on the activation of caspases was time dependent, reaching a maximum after 12 hr of stimulation, and then decreasing to a minimum after 72 hr. Treatment with melatonin also evoked mitochondrial membrane depolarization and permeability transition pore induction, which caused loss of mitochondrial staining by calcein, and increased cell death by apoptosis/necrosis as demonstrated by propidium iodide positive-staining of cells after 72 hr of stimulation. In addition, the exposure of cells to melatonin resulted in an activation and association of the proapoptotic proteins Bax and Bid, as well as promoting detectable increases in the expression of both proteins. We conclude that melatonin has proapoptotic and/or oncostatic effects in the human myeloid cell line HL-60.