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M De la Fuente

Publications and source records attributed to M De la Fuente.

At least 19 recordsLinked to original sources
PubMed2005

Modulation of neuropeptide Y and norepinephrine on several leucocyte functions in adult, old and very old mice.

The age-related changes in the communication between the nervous and the immune system have been scarcely investigated, especially in very aged subjects. The present work deals with the in vitro effects of norepinephrine and neuropeptide Y, separately and jointly, on functions such as lymphoproliferation, NK activity, and IL-2 and TNF-alpha release of peritoneal leucocytes from adult (24+/-2 weeks), old (72+/-2 weeks) and very old (128+/-2 weeks) mice. The old mice showed a decrease in proliferation, NK activity and IL-2 release, and an increase in TNF-alpha, whereas in the very old mice these functions were more similar to those of the adults. The effects of neurotransmitters on these functions were different depending on the age of the animals.

Aging↗
PubMed2004

Thiolic antioxidant supplementation of the diet reverses age-related behavioural dysfunction in prematurely ageing mice.

We have studied in a model of premature ageing in mice based on their impaired behavioural response in a simple T-maze test the effect of the ingestion of thioproline (TP) plus N-acetylcysteine (NAC) (0.1% w/w of each antioxidant) by female and male mice of Swiss and BALB/c strains on performance in two behaviour tests. The antioxidant treatment (4 weeks in two different periods of life, i.e., adult and old age) protected all animals against early-age-associated behavioural impairment, but this improvement was more evident in the prematurely ageing mice (PAM) in comparison to the control group or non-prematurely ageing mice (NPAM). An improvement of the exploratory activity and neuromuscular coordination after the thiolic antioxidant treatment was found in the PAM, bringing the behavioural parameters to the NPAM levels. These effects could be due to the glutathione precursor role of NAC and TP that replenish the intracellular reduced glutathione (GSH) levels despite advancing age. In conclusion, diet supplementation with thiolic compounds appears to be an effective therapy for protection against early behavioural decline in prematurely ageing mice.

Aging↗
PubMed2003

Several functions of immune cells in mice changed by oxidative stress caused by endotoxin.

We have studied natural killer (NK) activity, lymphoproliferative response, the release of several cytokines (IL-2, TNF alpha and IL-1 beta) and the ROS production in peritoneal leukocytes obtained 0, 2, 4, 12 and 24 h after lipopolysaccharide (LPS) injection. Lethal septic shock (100 % mortality occurred at 30 h after LPS administration) was caused in female BALB/c mice by intraperitoneal injection of 100 mg/kg of E. coli LPS. Cytotoxicity and lymphoproliferation assay were preformed together with the measurement of IL-1 beta, IL-2 and TNF alpha production, and quantification of ROS. Natural killer activity, spontaneous lymphoproliferative response, IL-2, TNF alpha, IL-beta release and ROS production were increased after LPS injection. In conclusions, ROS and proinflammatory mediators produced by immune cells in response to LPS are involved in the oxidative stress of endotoxic shock. This oxidative state alters some functional characteristics of leukocytes (proliferation and NK activity).

Animals↗
PubMed2002

Effects of antioxidants on immune system ageing.

One of the most widely accepted theories proposed to explain ageing is the free radical theory, according to which oxygen-derived free radicals cause age-related impairment through oxidative damage to biomolecules, with mitochondria being the main target of free radical attack. Since oxygen radicals are needed for many metabolic and physiological processes, an equilibrium between radical production and their antioxidant-linked inactivation is required to preserve health. Thus, senescence is the result of an imbalance between free radical production and antioxidant defences, with concomitant oxidative stress and age-dependent functional decline. This process is especially evident in the immune cells, which use free radicals in their functions and suffer a senescent deterioration probably linked to oxygen stress. Conversely, several laboratories, including our own, have shown that antioxidants preserve an adequate function of immune cells against homeostatic disturbances caused by oxidative stress, such as that involved with age. Therefore, since the immune system is an indicator of health and a longevity predictor, the protection of this system afforded by dietary antioxidant supplementation may play an important role in order to achieve a healthy ageing.

Aged↗
PubMed2002

Analysis of cDNAs from a mouse embryo tooth library: identification of novel genes during tooth development.

Signaling molecules, their receptors, and target genes from pathways and networks regulate the development of the tooth from initiation through cell differentiation. In order to identify genes involved in odontoblast and ameloblast differentiation, we constructed a cDNA library from E19.5 mouse molars. In this work, we report the partial cDNA sequences of 10 noncharacterized genes and we show cell expression of the transcripts on mouse embryo molars by in situ hybridization.

Animals↗
PubMed2002

Antibiotic resistance of Salmonella spp. from animal sources in Spain in 1996 and 2000.

Emergence of resistant and multiresistant bacteria has become an important worldwide sanitary problem. International agencies recommend improving resistance surveillance studies in not only human but also animal origin strains. Because of its ubiquitous characteristics and zoonotic agent consideration, Salmonella spp. can be used as a good indicator microorganism for resistance surveillance studies. Salmonella spp. strains from animal sources isolated in 1996 (107) and 2000 (474) in Spain were tested against 12 different antimicrobials agents, using the disc diffusion method. Results were interpreted following the NCCLS criteria. Data showed that Salmonella spp. strains (61.7% in 1996 and 81.5% in 2000) were resistant to at least one antibiotic. Pig-related strains were considerably more resistant than strains from other sources. Enteritidis serotype was less resistant than other serotypes, except for ampicillin in 1996 (50% resistant) and nalidixic acid in 2000 (65.1% resistant). An emergent monophasic serotype, 4,5,12:i:-, first detected in 1997 in Spain was 100% resistant and 90% multiresistant. Typhimurium serotype was the most common Salmonella serotype from animal sources in both years. It was widely distributed among animals and was among the serotypes with a higher degree of resistance. The ampicillin, chloramphenicol, sulfonamides, streptomycin, and tetracycline resistance pattern, commonly associated with Salmonella serotype Typhimurium DT 104, had spread among other Typhimurium phage types and other Salmonella serotypes. Salmonella spp. strains isolated from feeding stuffs were considerably more susceptible than animal source strains, suggesting that the high Salmonella spp. resistance percentage was probably due to the use of antibiotics in animal farms rather than the consumption of contaminated feeding stuffs.

Animals↗
PubMed2001

Neuropeptide Y effects on murine natural killer activity: changes with ageing and cAMP involvement.

Changes in the bidirectional interaction between the nervous and the immune systems have been proposed as a cause of ageing. Neuropeptides, such as neuropeptide Y (NPY), could show different effects on immune function with age. In the present work, we have studied the in vitro action of a wide range of NPY concentrations, i.e. from 10(-13) to 10(-7) M, on natural killer (NK) activity, a function which decreases with age. Spleen, axillary nodes, thymus and peritoneum leukocytes from mice of different ages: young (12+/-2 weeks), adult (24+/-2 weeks), mature (50+/-2 weeks) and old (72+/-2 weeks) were used. Stimulation by NPY of NK activity was observed in adult and mature animals in axillary nodes and thymus, and an inhibition in the spleen from young mice. The specificity of the NPY effect on cytotoxic activity was confirmed using a C-terminal fragment of NPY. Furthermore, cAMP levels in leukocytes were found to be decreased by NPY in adult mice, suggesting an involvement of this messenger system in the NK modulation by this neuropeptide.

Aging↗
PubMed2001

Changes with aging in the modulation by neuropeptide Y of murine peritoneal macrophage functions.

Some age-related changes in immune function may be due, at least in part, to a disturbance in the communication between the nervous and immune systems. In the present work, the effects in vitro of neuropeptide Y (NPY) (10(-13) to 10(-7) M) on different peritoneal macrophage functions (adherence to substrate, chemotaxis, phagocytosis, superoxide anion production, and the release of TNFalpha and IL-1beta) have been studied on cells from young (12+/-2 weeks), adult (24+/-2 weeks), mature (50+/-2 weeks) and old (72+/-2 weeks) BALB/c mice. The specificity of these actions was confirmed using two C-terminal fragments of NPY, and the intracellular messengers (protein kinase C and cAMP) involved in the action of the neuropeptide were also analyzed. The results show that the functions studied change with aging and that the effects of NPY on each function, which are carried out through specific receptors, as well as on intracellular pathway, differ depending on age, maintaining the immune functions at physiologically adequate levels in old animals.

Aging↗
PubMed2001

Behavioral characterization of a mouse model of premature immunosenescence.

In previous studies we have shown that differences in life span among members of Swiss mouse populations appear to be related to their performance in a T-maze, with a slow performance ("slow" mice) being linked to an impaired immune function and a shorter life span when compared to "fast" mice, which led us to propose the slow mice as a model of immunosenescence. In the present study we demonstrate that in a tightrope test of neuromuscular vigor and coordination the slow mice show a worse performance, needing more time to complete the task. Moreover, these animals show a decreased locomotor activity and an increased level of emotionality/anxiety in three standard behavioral tests (the holeboard, the open field and the plus-maze) when compared to fast mice. All these behavioral features were most marked in the slow females. The results also indicate that slow animals show a decreased chemotaxis of macrophages and lymphocytes, as well as a reduced lymphoproliferative response to mitogens. The data supports our claim that slow or hyperemotional mice, in which immune and neurobehavioural functions appear to be impaired, may be a useful model of premature aging.

Aging, Premature↗
PubMed2001

The immune response of thymic cells from the turtle Mauremys caspica.

In the present work several known mammalian leukocyte functions such as directed mobility, proliferative response to mitogens, antibody-dependent cellular cytotoxicity (ADCC) and natural killer (NK)-mediated cytotoxicity were studied in female and male Mauremys caspica turtles. Since the reptilian thymus shows seasonal variations in its structure, we have performed all the assays along the seasonal cycle. Our results show that thymic cells from M. caspica are able to migrate through a chemo-attractant gradient, to proliferate in response to the mitogens phytohaemagglutinin (PHA) and concanavalin A (Con A), and to kill tumoral target cells by both ADCC-mediated and NK-mediated cytotoxicity. Those functions were differentially affected by the seasonal cycle; in general, in autumn the functions studied showed the smallest values for both sexes, while in summer the highest values of cytotoxicity and chemotaxis were found in females. The proliferative responses to PHA and Con A were higher for both sexes in spring and for females in winter than in the other seasons. In summary, thymic cells from M. caspica show a wide range of immune functions, and these are modulated heterogeneously by the seasonal cycle in both sexes.

Animals↗
PubMed2001

Ascorbic acid and N-acetylcysteine improve in vitro the function of lymphocytes from mice with endotoxin-induced oxidative stress.

Oxidative stress associated with reactive oxygen species (ROS) and cytokines produced by immune cells, which is involved in septic shock caused by endotoxin, can be controlled to a certain degree by antioxidants with free radical scavenging action. N-acetylcysteine (NAC) and ascorbic acid (AA) are ROS scavengers that improve the immune response, and modulate macrophage function in mice with endotoxin-caused oxidative stress. Therefore, we have investigated the in vitro effects of these antioxidants on the functions of lymphocytes from BALB/c mice with lethal endotoxic shock caused by intraperitoneal injection of E. coli lipopolysaccharide (LPS) (100 mg/kg). Adherence to tissues and chemotaxis (the earliest two functions of lymphocytes in the immune response), as well as ROS levels and TNF alpha production were determined in the presence or absence of NAC or AA (0.001, 0.01, 0.1, 1 and 2.5 mM) in lymphocytes from peritoneum, axillary nodes, spleen and thymus obtained at several times (2, 4, 12 and 24 hours) after LPS injection. Endotoxic shock decreases the chemotaxis of lymphocytes from all the above localizations and increases their adherence, TNF alpha and ROS production. These changes in lymphocyte function were counteracted by NAC and AA, bringing these functions to values near those of control animals. Our data suggest that lymphocytes are important targets of endotoxins contributing to oxidative stress by septic shock, and that antioxidants can preserve the function of lymphocytes, preventing the homeostatic disturbances caused by endotoxin.

Acetylcysteine↗
PubMed2001

Intracellular glutathione deficiency is associated with enhanced nuclear factor-kappaB activation in older non-insulin dependent diabetic patients.

Diabetes mellitus may be associated with intracellular glutathione (GSH) deficiency. Since in vivo studies have shown that plasma intracellular GSH plays a key role in regulating the activation of nuclear factor kappaB (NF-kappaB), we have investigated the relationship between intracellular thiols (GSH, homocysteine, cysteine and cysteinyglycine) and NF-kappaB activity in the peripheral blood mononuclear cells (PBMC) of 63 elderly non-insulin dependent diabetes mellitus (NIDDM) patients (28 microalbuminurics and 35 normoalbuminurics) and 30 healthy age- and sex-matched subjects. In addition, we have measured plasma concentrations of these thiol compounds, serum concentrations of interleukin-6 (IL-6) and vascular cell adhesion molecule-1 (sVCAM-1), that are partly dependent on the NF-kappaB activation, as well as the serum levels of thiobarbituric acid reacting substances (TBARS), as index of lipid peroxidation. Diabetic patients with microalbuminuria (MAB) and normoalbuminuria had NF-kappaB activity 2.1- and 1.5-fold greater, respectively, than the control group. As compared to normoalbuminuric patients, patients with MAB had significantly higher levels of glycemia, plasma homocysteine, and serum concentrations of TBARS, IL-6 and sVCAM-1 (in all cases, p < 0.01), and significantly lower GSH content in the PBMC (p < 0.05). The intracellular GSH in PBMC correlated with NF-kappaB activation (r = -0.82; p < 0.0001), serum TBARS (r = -0.60; p < 0.001), and with fasting glycemia (r = -0.56; p < 0.001) in patients with MAB, whereas a weaker association between GSH levels in PBMC and NF-kappaB activation (r = -0.504, p < 0.001) was seen in patients without MAB. These results suggest that the decrease of intracellular GSH content in elderly NIDDM patients with MAB is strongly associated with enhanced NF-kappaB activation, which could contribute to the development of increased glomerular capillary permeability and its rapid progression.

Age Factors↗
PubMed2001

Changes in the ascorbic acid levels of peritoneal lymphocytes and macrophages of mice with endotoxin-induced oxidative stress.

Ascorbic acid (AA) is an important cytoplasmic antioxidant that mice synthesize in the liver, the intracellular levels of which decrease in an oxidative stress situation such as endotoxic shock. The present work deals with the changes in AA levels, that modulate the immune function, in the two main immune cells, namely macrophages and lymphocytes, from female BALB/c mice suffering endotoxic shock caused by intraperitoneal injection of Escherichia coli lipopolysaccharide (LPS) (100 mg/kg). The intake by cells of this antioxidant present in vitro at different concentrations was also studied. The animals show an oxidative stress, standardized in previous studies, that causes mortality at 30 h after LPS injection. The cells were obtained from the peritoneum at 2, 4, 12 and 24 h after LPS or PBS (control) injections and were incubated without or with AA at 0.01, 0.1 and 1 mM for 10, 30, 60, 120 or 180 min. The hepatic AA levels were also studied at 0, 2, 4, 12 and 24 h after LPS injection. The peritoneal cells obtained from animals injected with LPS showed increased AA levels in relation to the control cells at all times after LPS injection, with maximal effect at 12h. The AA levels decreased after this time, in agreement with changes in the AA hepatic levels. The increase was due to the AA of lymphocytes since macrophages showed a decrease in AA at different times after LPS injection. Both cells showed an increase in the intracellular levels of AA when this antioxidant was added in vitro. This takes place mainly at 30-60 min of incubation in cells from controls and at 10 min in cells from treated mice 12-24 h after LPS injection. The incorporation decreased at these times of endotoxic shock, a few hours before death. In all cases AA levels were higher in lymphocytes than in macrophages, and 1 mM was the most effective concentration. These results suggest that the immune cells need appropriate levels of antioxidants, such as AA, under oxidative stress conditions, and that while lymphocytes take and accumulate AA, macrophages use it.

Animals↗
PubMed2000

The NPY effects on murine leukocyte adherence and chemotaxis change with age. Adherent cell implication.

The two-way communication between the nervous and immune system is currently well-known, but the age-related changes in this communication have been scarcely studied. In the present work, we have investigated the in vitro effects of neuropeptide Y (NPY) at concentrations ranging from 10(-13) to 10(-7) M on the adherence and chemotaxis capacities of spleen, axillary node, thymus and peritoneum leukocytes from BALB/c mice. The NPY effect on these functions was examined on cells from animals of four different ages, i.e. young (12+/-2 weeks old), adult (24+/-2 weeks old), mature (50+/-2 weeks old) and old (72+/-2 weeks old). In young animals, NPY stimulates the adherence of leukocytes from spleen, axillary nodes and thymus and inhibits it in cells from peritoneum. In adult animals NPY inhibits the adherence of leukocytes from thymus. These effects disappear with ageing in all locations. Chemotaxis is stimulated by this neuropeptide at all ages in cells from axillary nodes and peritoneum, but this effect is absent in old mice. NPY exerts an inhibitory effect on the chemotaxis of leukocytes from thymus at all ages studied. These NPY effects on leukocytes seem to be carried out through adherent cells.

Aging↗
PubMed2000

Seasonal changes in peripheral blood leukocyte functions of the turtle Mauremys caspica and their relationship with corticosterone, 17-beta-estradiol and testosterone serum levels.

We have studied the most representative functions of lymphocytes such as adherence to substrate, mobility directed to a chemoattractant gradient (chemotaxis), proliferative response to mitogens and antibody-dependent cellular cytotoxicity (ADCC), as well as natural killer (NK) activity in peripheral blood cells from the turtle Mauremys caspica, and the seasonal changes of these functions in both female and male animals. The plasma levels of steroid hormones were determined to study their relationship with the immune functions. Our results show high chemotaxis, lymphoproliferative response and ADCC as well as NK activity in winter when the levels of corticosterone, testosterone and 17-beta-estradiol were depleted. Proliferative responses to phytohaemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM) were increased in spring correlating with low levels of corticosterone and testosterone in middle and late spring. In summer, the proliferative response was decreased but adherence, chemotaxis, ADCC and NK activity were increased, although steroid hormones showed high plasma levels. In autumn, a depletion of both the hormone levels and the immune response were found except for adherence to substrate. The immune functions studied were strikingly influenced by the seasonal cycle, which induced a different pattern of response depending on the function analyzed. Moreover, these immune cells showed a different degree of dependence on steroids in relation to the function and the season considered, suggesting the existence of other factors that modulate the immune response studied.

Animals↗
PubMed2000

Changes with aging in the modulation of macrophages by norepinephrine.

The effect of aging on the norepinephrine (NE)-induced modulation of phagocytic and oxygen-dependent microbicidal processes of mouse peritoneal macrophages was studied. Phagocytosis of latex beads on culture plates and superoxide anion production was evaluated in young (12 weeks), adult (22 weeks), mature (48 weeks) and old (72 weeks) BALB/c mice after in vitro incubation with 10(-12), 10(-9), 10(-7), 10(-5) or 10(-3) M concentrations of NE. The results indicate that the phagocytic response to NE is quite similar in young and mature mice, with increased phagocytosis after incubation with 10(-7) and 10(-3) M, and decreased phagocytosis with 10(-5) M. Macrophages from adult mice increased their phagocytic capacity after incubation with the highest concentrations of NE (10(-5) and 10(-3) M) and macrophages from old animals only were stimulated with 10(-3) M. In addition, it was found that usually NE increased the extracellular superoxide anion production in the absence of phagocytosis in adult mice. No statistically significant changes were found in intracellular superoxide anion levels, but an increase was seen after phagocytosis in macrophages from adult, mature and old animals, especially after incubation with 10(-5) M. In conclusion, the results obtained indicate that the modulation of macrophages by NE does not only depend on the concentration of this neurotransmitter, but also on age.

Aging↗
PubMed2000

Effect of aging on the modulation of macrophage functions by neuropeptides.

The existence of a functional connection between the nervous and the immune system is supported by increasing recent evidence. In previous work we have shown that peptides from the nervous system, such as gastrin-releasing peptide (GRP), neuropeptide Y (NPY) and sulfated cholecystokinin octapeptide (CCK-8s), have modulatory effects on the immune functions in adult animals. Since the immunodepression found in aging organisms may be related to changes in the neuroimmune network, the aim of the present work was to study the changes with aging in the effect of CCK-8s, GRP and NPY on peritoneal macrophage functions (adherence to tissues, mobility, ingestion of foreign particles and superoxide anion production) from BALB/c mice of three different ages: adult (24+/-2 weeks old), mature (50+/-2 weeks old) and old (72+/-2 weeks old). The results show that the increase in adherence capacity produced by neuropeptides in cells from adult and mature animals disappears in old mice. The stimulatory effect of GRP and NPY on mobility, ingestion and superoxide production in macrophages from adult mice disappears (GRP) or changes to inhibition (NPY) in cells from old animals. The decrease of these functions caused by CCK-8s in adult or mature animals continues in old mice. These data suggest that the modulation by neuropeptides of the macrophage function changes with the age of animals.

Aging↗