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Biomedical subjects

J M Peinado

Publications and source records attributed to J M Peinado.

At least 19 recordsLinked to original sources
PubMed1999

A mathematical model for toxin accumulation by killer yeasts based on the yeast population growth.

The accumulation of toxin by killer yeast populations is modelled starting from a mechanistic approach that explains the toxin production in terms of yeast population growth, and takes into account the environmental inactivation of the toxin. A modified Richard's general equation for limited growth is used to define the function that describes the toxin produced in relation to the yeast biomass increase. The relationship between the rates of cell and toxin production is explicitly shown, and the implications of the resulting proportionality factor are discussed. The model parameters have been adjusted and the model has been validated using experimental data of growth and toxin accumulation from cultures of Pichia membranaefaciens in two different media. The differences between both types of cultures are analysed on the basis of parameter estimates and the predicted rate of toxin production per cell. The results support the hypothesis that biomass production and toxin synthesis are controlled in different ways; they also suggest that the composition of the medium could have a distinct effect on toxin synthesis. Model assumptions are discussed in comparison with a previous model for killer-sensitive interaction of Saccharomyces cerevisiae strains.

Computer Simulation↗
PubMed1999

A differential medium for the isolation of Kluyveromyces marxianus and Kluyveromyces lactis from dairy products.

A selective and differential solid medium, called Kluyveromyces Differential Medium (KDM), is described for the isolation of Kluyveromyces marxianus and K. lactis from dairy products. Its discriminative potential is based on the detection of the enzyme beta-galactosidase, in the absence of lactose. Of the more than 95 strains tested, including yeasts, bacteria, and filamentous fungus, only the strains of K. marxianus and K. lactis produced blue colonies on the medium due to the presence of X-Gal/ IPTG. The bacterial strains were not able to grow in KDM. On this basis, the medium was very satisfactory when testing naturally or experimentally contaminated dairy food products. When quality assessment tests were performed, optimal values of productivity (growth and color) and selectivity were obtained for K. marxianus and K. lactis.

Animals↗
PubMed1998

Quantitative and ultrastructural changes in glia and pericytes in the parietal cortex of the aging rat.

The frequency of astrocytes, microglia plus oligodendrocytes, and pericytes displaying nuclei was analyzed and quantified in 160-microm-wide strips of the parietal cortex (Par1 region) from young and aged Wistar rats. The study was performed on two groups of rats aged 3-4 and 32-36 months. Quantifications of the glial cell types and pericytes were made in 1-microm-thick sections stained with toluidine blue. Ultrathin sections were also made to analyze the ultrastructural features of these cells during aging. Astrocytes and pericytes increased in number by about 20% and 22%, respectively, with age. These increases were most significant in layers II-IV and V for both cellular types. Clusters of astrocytes were common in these layers of aging rats. The ultrastructural analysis also indicated changes in all cell types that stored inclusions and vacuoles with age, which were particularly abundant in microglial cells. End-feet astrocytes and pericytes surrounding the vascular wall also contained vacuoles and inclusions, and consequently the vascular wall increased in thickness. In conclusion, the aging process increased astrocyte and pericyte populations, but not microglia plus oligodendrocyte populations, in the rat parietal cortex. Although no significant change in nuclear size could be observed in any cell type, all glial cells as well as pericytes underwent morphological ultrastructural changes. These modifications may result from the need to correct possible homeostatic imbalances during aging.

Aging↗
PubMed1998

Evaluation of carnitine nutritional status in full-term newborn infants.

Carnitine supplements may be advisable not only in premature but also in artificially-fed full-term babies. The acyl-carnitine/free carnitine (AC/FC) and FC/total carnitine (FC/TC) ratios have been considered markers of "carnitine insufficiency" and "carnitine deficiency", respectively. Values of AC/FC>0.40 are considered abnormal and mean that FC has a low bioavailability to the cells and so reflects a "carnitine insufficiency". Values of FC/TC<0.7 indicate "carnitine deficiency". We analyze the validity of such ratios and the limits for them in three groups of full-term neonates (n=66): 22 breast-fed (BF), 22 with formula (F); and 22 fed with carnitine-supplemented formula. Several studies have shown the need to give supplements of carnitine to the neonate because of its "essentiality", but no one has demonstrated the adequate dosages. We therefore propose to establish new limit levels for these ratios to control carnitine nutritional status in neonates, based on the control of percentile ranges for normal BF infants (in this study: 97th percentile of AC/FC>0.83; 3rd percentile of FC/TC<0.54) and on evaluating the needs of neonates and dosages required to supplement F. The supplement of 2.2 mg of L-carnitine/100 ml in the cow's milk formula used in the present study produces a similar biochemical pattern of plasma carnitine and ACs to that observed in BF infants, together with a lower risk of developing "carnitine deficiency" or "carnitine insufficiency" than those babies fed with nonenriched F. Considering that human milk is the best source of nutrition for full-term infants, the limit established for AC/FC and FC/TC ratios at other ages of life seems to be "inadequate" for neonates.

Carnitine↗
PubMed1997

Light microscopic quantification of morphological changes during aging in neurons and glia of the rat parietal cortex.

BACKGROUND: Different changes in neuronal and glial population of the aging brain have been described; however, the degree and extent of these changes are controversial. This study evaluates the quantitative and cytomorphometric effects of aging on neuronal and glial populations in the parietal cortex of the rat. METHODS: The study was performed in two groups of rats aged 4-6 and 30-32 months. Cortical volume, neuronal density, glial density, and neuronal area, and shapes of the soma and nucleus were analyzed in cortical layers I, II-IV, V, and VI using serial sections stained with cresyl-fast-violet, and quantitative morphometric techniques. RESULTS: No changes with age were found in volume of the cortex or neuronal density. Glial density increased significantly (mean for all layers 17%) in older rats. Layers II-IV, V, and VI showed an age-related decrease in the area of the neuronal soma. Neuronal shape, as revealed by the major/minor diameter ratio, also showed a decrease in old rats but only in layer II-IV. Nuclear area decreased with age only in layer VI. CONCLUSIONS: The stability of neuronal density together with the increased number of glial cells and the changes in neuronal soma size suggest that aged-related cognitive impairment could be a consequence of neuronal dysfunction rather than actual neuronal losses.

Aging↗
PubMed1997

Evoked GABA release is not mediated by N-type VDCC in the frontal cortex of awake rats: effects of neomycin.

The purpose of the present study was to analyze the Ca2+ channel involved in GABA release under resting and K(+)-evoked conditions in vivo. We used microdialysis to investigate the effects of the voltage-dependent calcium channel (VDCC) blockers neomycin, kanamycin, and omega-conotoxin GVIA, and the voltage-dependent Na+ channel blocker tetrodotoxin, in the frontal cortex of awake rats. The GABA content in frontal dialysates was analyzed by high performance liquid chromatography coupled to fluorescence detection. Basal GABA release was kanamycin, omega-conotoxin, and tetrodotoxin resistant, whereas neomycin induced a significant increase from the basal level. The K(+)-evoked release of GABA was kanamycin and omega-conotoxin resistant, but tetrodotoxin sensitive. The effects of neomycin were masked by the action of this drug on basal release. These results suggest that neomycin may affect GABA release in the frontal cortex through a mechanism independent of VDCC. In addition, the K(+)-evoked release of GABA in this cortical area was not mediated by the N-type voltage-dependent calcium channels, but was dependent on neural activity or TTX sensitive.

Animals↗
PubMed1997

Effect of salt on the killer phenotype of yeasts from olive brines.

The killer properties of yeasts isolated from olive brines were examined in the absence and presence of sodium chloride in concentrations of up to 6% (wt/vol). An apparent enhancement of the killing action as the salt concentration increased, as well as changes in the spectra of activity against selected target strains, was observed in a few strains. Culture filtrates from killer strains grown at different NaCl concentrations (0, 3, or 6% [wt/vol]) were tested against sensitive yeasts cultivated under the same conditions. While the sensitivity of the target strain greatly increased in the presence of salt, no significant effect on toxin production was noticed.

Dose-Response Relationship, Drug↗
PubMed1997

[Age-related changes in the cytolytic activity of aspartyl aminopeptidase in the cerebral cortex of the rat].

INTRODUCTION: Recently, neurotoxicity mediated by excitatory amino acids has been considered to be the etiopathogenic basis of various neurodegenerative processes, including aging of the brain. OBJECTIVE: Since the enzyme aspartyl-aminopeptidase (Asp-AP) may play a role in modifying the concentration of the cytotoxic amino acids glutamate and aspartate, we studied the changes in its activity in the rat cerebral cortex, which were possibly related to age. MATERIALS AND METHODS: Homogenates of frontal, parietal, temporal and occipital cortex were obtained from young and old rats. The activity of Asp-AP in these homogenates determined by fluorimetry. RESULTS: Our results showed a significant age-dependent drop in Asp-AP activity in the frontal, parietal and occipital cortices, with little change in the temporal cortex. CONCLUSIONS: These results may be understood as part of a general schedule of age-dependent modifications of the cerebral protease systems.

Aminopeptidases↗
PubMed1996

The inactivation of hexokinase activity does not prevent glucose repression in Candida utilis.

High hexokinase activity was not related to glucose repression in Candida utilis IGC 3092. The addition of Cibacron Blue 3G-A to growing cells in batch culture led to a permanent in vivo hexokinase inactivation, decreased growth rate and inhibited alcohol dehydrogenase. Hexokinase inactivation up to 90% did not alleviate glucose repression of alpha-glucosidase, as has been described for Saccharomyces cerevisiae and other yeasts. Moreover, when cells were physiologically derepressed by growing them in a chemostat at low glucose concentrations, the highest hexokinase activity was shown by the derepressed cells, and decreased as repression increased. Thus, in our strain of C. utilis, hexokinase activity was inversely proportional to glucose repression.

Candida↗
PubMed1995

Decrease in cytosolic Aspartyl-aminopeptidase but not in Alanyl-aminopeptidase activity in the frontal cortex of the aged rat.

To test the neurotoxic hypothesis of excitatory amino acids, we evaluated the possible contribution to the free acidic amino acid pool of Aspartyl-aminopeptidase activity in the frontal cortex of adult (3 month old) and aged rats (3 groups of animals aged 26, 29 and 33 months). Aspartyl-aminopeptidase activity showed a significant decrease in the oldest rats (29 and 33 months old) whereas the activity of Alanyl-aminopeptidase, an unspecific enzyme, did not change with age. These data invalidate the idea that excess free acidic amino acids are released by aminopeptidases in the aged rat but do provide evidence of age-related changes in this enzymatic activity. The possible implications of our findings for general alterations in protein degradation are discussed.

Aging↗
PubMed1993

Quantitative morphological changes in neurons and glia in the frontal lobe of the aging rat.

The quantitative morphological changes in neurons and glia during the aging process were analyzed in the different cortical layers, grouped as I, II-IV, V, and VI, of the frontal cortex of the rat. The parameters analyzed were cortical volume, neuronal density, glial density, and neuronal soma and nucleus areas. No changes with age were found in the volume of the layers, in neuronal density (with the exception of layer I), or in the area of the neuronal soma. However, older animals showed a 10 to 20% increase in glial density, depending on the layer studied. In addition, there was an age-related decrease in the area of the neuronal nucleus in layers II-IV, V, and VI. These results support the idea that the aging frontal cortex undergoes structural changes that may be involved in the morphological basis of memory and cognitive impairments characteristic of aging.

Aging↗
PubMed1989

Profile of NE, DA and 5-HT activity shifts in medial hypothalamus perfused by 2-DG and insulin in the sated or fasted rat.

This study was carried out in the unrestrained rat to determine the nature of the in vivo profile of monoamine neurotransmitters within the medial hypothalamus in response to the presence of a glucoprivic or metabolic challenge to neurons within this region. In these experiments, insulin or 2-deoxy-D-glucose (2-DG) was applied locally to the paraventricular nucleus (PVN), dorsomedial nucleus (DMN) and ventromedial hypothalamus (VMH). In each of 11 Sprague-Dawley rats, a guide cannula was implanted stereotaxically to rest just above these structures. Upon recovery, a concentric push-pull cannula system was used to perfuse an artificial CSF within a medial hypothalamic site. The CSF was perfused at a rate of 20 microliters/min with a 5.0 min interval intervening between the collection of each 100 microliters sample. After the rat was fasted for 20-22 hr, either 10 micrograms/microliters 2-DG or 4.0 mU/microliters of insulin was incorporated into the control CSF medium and perfused at the same locus. The aliquots of hypothalamic perfusate were assayed by high performance liquid chromatography with electrochemical detection (HPLC-EC) for the respective concentration in pg/microliter of norepinephrine (NE), dopamine (DA), serotonin (5-HT) and each of their major metabolic products. When the rat was sated, 2-DG enhanced significantly the mean efflux of NE from the medial hypothalamus in comparison to control CSF values. However, under the fasted condition, 2-DG augmented the turnover of both the catecholamine and 5-HT as reflected by elevated levels of MHPG and 5-HIAA, respectively. On the other hand, insulin perfused within the same medial hypothalamic sites evoked a significant increase in the synthesis and release of DA from the sated rat, but did not alter its turnover. Following the interval of fast, insulin produced no immediate alteration in transmitter activity; however, in the interval following insulin's perfusion, DA and 5-HT turnover were enhanced while the efflux of 5-HT was suppressed. An analysis of the proportional values of the levels of the amines to each other revealed marked shifts in the relationships between the catechol- and indoleamine transmitters following local perfusion with both 2-DG and insulin. Overall, NE synthesis and turnover exceeded that of 5-HT following 2-DG, whereas DA predominated over NE and 5-HT during insulin's perfusion.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1989

Regulation of glucose transport in Candida utilis.

The transport systems for glucose present in Candida utilis cells, growing in batch and continuous cultures on several carbon sources, have been studied. Two different systems were found: a proton symport and a facilitated diffusion system. The high-affinity symport (Km for glucose about 15 microM) transported one proton per mole of glucose and was partially constitutive, appearing in cells grown on gluconeogenic substrates such as lactate, ethanol and glycerol. It was also induced by glucose concentrations up to 0.7 mM and repressed by higher ones. The level of repression depended on the external glucose concentration at which cells had grown in a way similar to that shown by the maltose-uptake system, so both systems seem to be under a common glucose control. Initial uptake by facilitated diffusion, the only transport system present in cells growing at glucose concentrations higher than 10 mM, showed a complex kinetic dependence on the extracellular glucose concentration. This could be explained either by the presence of at least two different systems simultaneously active, one with a Km around 2 mM and the other with a Km of about 1 M, or by the allosteric or hysteretic behaviour of a single carrier whose apparent Km would oscillate between 2 and 70 mM.

Biological Transport↗
PubMed1988

Monoamine transmitter activity in lateral hypothalamus during its perfusion with insulin or 2-DG in sated and fasted rat.

A unique profile of neurochemical events is proposed to occur in the diencephalon which is contingent upon the nutrient status of the animal. In this first of a series of investigations, we selected the lateral hypothalamus (LH) in order to determine its specific resting profile of monoaminergic neurotransmitters and their principal metabolites. The neuronal pattern of activity was studied during sated and fasted conditions as well as during a local glucoprivic challenge to the LH. After permanent guide cannulae for push-pull perfusion were implanted in female Sprague-Dawley rats, the LH was perfused repeatedly with an artificial CSF, at a rate of 20 microliters/min, in order to collect a series of 5.0 min samples. Aliquots of each perfusate were assayed directly using a high performance liquid chromatography system with electrochemical detection (HPLC-EC) for pg/microliter concentrations of norepinephrine (NE), dopamine (DA) and serotonin (5-HT). In comparison to the basal levels of amines during the sated condition, when the rat was food-deprived for 20-22 hr, the release of NE, DA, and 5-HT was significantly lower than that observed under the sated condition. Further, the turnover of NE in the LH was concurrently attenuated as reflected by the lower levels of MHPG in the perfusate, thus demonstrating the modification in catecholamine activity produced in the LH by the condition of hunger. When either 10 micrograms/microliters 2-deoxy-D-glucose (2-DG) or 4.0 mU/microliter insulin was incorporated into the CSF perfused in the LH, the efflux of DA was significantly enhanced independent of the state of satiation. In addition, the proportion of both NE and DA to 5-HT was likewise increased by either of these centrally acting substances, while the turnover of 5-HT was enhanced and NE and DA turnovers were reduced. Perfusion of 2-DG in the LH of the fasted rat caused a significant reduction in catecholamine turnover in terms of MHPG/NE, VMA/NE, DOPAC/DA and HVA/DA ratios. Moreover, 2-DG increased NE/5-HT while lowering the NE/DA ratio, and enhanced simultaneously the 5-HTOL/5-HT ratio. In the sated rat, 2-DG attenuated the release of 5-HT from the animal's LH, whereas insulin caused a shift in the proportions of NE/5-HT and DA/5-HT. Further, the peptide served to reduced the efflux of 5-HT, enhanced the turnover of 5-HT while diminishing DA turnover, and shifted the metabolism of NE from MHPG to VMA.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

In vivo analysis of cortical amino acid neurotransmitters collected in the rat by a new double lumen push-pull catheter system.

The release of both endogenous and newly synthesized amino acid neurotransmitters was examined simultaneously in different areas of the cerebral cortex in the freely moving rat. An array of push-pull guide tubes was implanted permanently to rest above the frontal, parietal, temporal and occipital areas of the cortex of each rat. Then a new double-lumen catheter system, specially adapted for localized push-pull perfusion of the conscious animal, was used to perfuse an artificial cerebrospinal fluid at each cortical site. For the new synthesis experiments, 0.5 microCi of [14C]glucose in a volume of 2.0 microliter was first microinjected into the perfusion site as a precursor to label amino acids. After the site was perfused at a rate of 12.0 microliter/min, each of the samples was assayed by two-dimensional thin-layer chromatography. In a second analysis, the content of six endogenous amino acids present in unlabeled samples of push-pull perfusate was quantified by high-performance liquid chromatography analysis with electrochemical detection. The results showed a notable homogeneity among each of the four cortical areas in the content of four of the six amino acids examined. Endogenous glutamine exhibited the highest proportional content in the cortical perfusates, whereas glutamic acid was proportionally higher in terms of new synthesis. An anatomical analysis revealed that the level of endogenous glutamic acid in the frontal area was significantly lower than that found in the occipital or temporal regions of the rat's cortex. An opposite result was obtained when the proportional synthesis of glutamic acid from [14C] glucose was compared in different cortical regions in that a statistically higher release occurred in the frontal than in the occipital cortex.

Amino Acids↗