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Biomedical subjects

M A García

Publications and source records attributed to M A García.

At least 37 records · Page 2Linked to original sources
PubMed2003

The latency protein LANA2 from Kaposi's sarcoma-associated herpesvirus inhibits apoptosis induced by dsRNA-activated protein kinase but not RNase L activation.

Kaposi's sarcoma-associated herpesvirus (KSHV) uses several strategies to counteract the interferon (IFN) system. In this study, the relationship of the protein LANA2 from KSHV to the IFN-activated protein kinase (PKR) and 2-5A system was analysed. It was found that LANA2 could not abrogate apoptosis or RNA degradation mediated by the 2-5A system. However, expression of LANA2 inhibited apoptosis triggered by PKR. LANA2 also counteracted the PKR-mediated inhibition of protein synthesis and partially blocked PKR-induced phosphorylation of eIF-2alpha. Analysis of PKR-induced activation of caspases 3 and 9 revealed that LANA2 abrogated activation of caspase 3 but not of caspase 9. These findings show that LANA2 is able to interfere with downstream events triggered by PKR. Hence, LANA2 should be considered as a KSHV defence protein against IFN.

2',5'-Oligoadenylate Synthetase↗
PubMed2002

Soil CO2 fluxes in cereal land use of the Spanish plateau: influence of conventional and reduced tillage practices.

From November 1998 to October 2000, measurements of soil respiration were performed on the Spanish plateau for two patches of non-irrigated barley, one managed with conventional tillage (CT) and the other with reduced tillage (RT). Soil CO2 flux showed seasonal variation on both patches, with an increase from March to October, peaking in May, and a decrease during the winter period by a factor of around 2. The mean value for both combined years was 2.03 and 1.70 micromol m(-2) S(-1), in the CT and RT patches, respectively. In order to analyse the influence of RT on soil CO2 flux, two tests were performed. The first one was the Kruskal-Wallis test to compare whether the differences between the medians in both patches were statistically significant. The results obtained revealed statistically significant differences during the second year, at a 85% and 95% significance level, use being made of annual data and that recorded during the period of maximum interest, March-October, respectively. The decrease in soil respiration in the RT patch was around 24%. The second test was aimed at describing and comparing the influence of soil temperature on soil CO2 flux. By using the data of both patches recorded during the first year, an empirical equation on 10-cm soil temperature was fitted and tested on the data corresponding to the second year in each of the patches. Then, a comparison between the medians of the differences between the estimated and observed values was again performed by means of the Kruskal-Wallis test. The over-prediction of the model in the RT patch, statistically significant at a 90% significance level, was roughly 23%, confirming again the decrease in soil respiration one year after this agricultural management practice had been implemented.

Agriculture↗
PubMed2002

The effect of dietary protein content and feeding level on the rate of protein deposition and energy utilization in growing Iberian pigs from 15 to 50 kg body weight.

The effects of dietary protein content and feeding level on the utilization of metabolizable energy (ME) and on the rates of gain, protein and fat deposition have been studied in seventy-two Iberian pigs growing from 15 to 50 kg body weight (BW) by means of comparative slaughter experiments. The animals were fed on six diets providing 223, 192, 175, 156, 129 and 101 g crude ideal protein ( CP)/kg DM and 14.64, 14.14, 14.37, 14.80, 15.36 and 15.53 MJ ME/kg DM respectively. Each diet was offered at three levels of feeding: 0.60, 0.80 and 0.95xad libitum intake. Protein deposition (PD) increased significantly (P<0.01) with each decrease in dietary CP content and reached a maximum value (74.0 g) when the diet providing 129 g CP/kg DM (6.86 g digestible ideal protein/MJ ME) was offered at the highest feeding level. This feeding regimen resulted in average values for live-weight gain and retained energy (RE) of 559 g/d and 10.9 MJ/d respectively. RE increased significantly (P<0.001) from 480 to 626 kJ/kg BW0.75 with each decrease in dietary CP content from 192 to 129 g/kg DM. Raising the level of feed intake led to significant linear increases in PD and RE irrespective of the diet fed (P<0.001). When diets approaching an adequate supply of CP were given, the net efficiency of use of ME for growth (kw) and the maintenance energy requirements were 58.2 % and 422 kJ/kg BW0.75 per d respectively.

Adipose Tissue↗
PubMed2002

Pharmacokinetics and ocular penetration of grepafloxacin in albino and pigmented rabbits.

The pharmacokinetics of grepafloxacin were determined in albino and pigmented rabbits following a single 10 mg/kg intravenous administration. The penetration of grepafloxacin into various ocular tissues was also determined after continuous intravenous infusion in both types of animal. Grepafloxacin showed a bicompartmental model of distribution in both pigmented and albino rabbits with significant differences in the pharmacokinetics between the two types of animal. After continuous intravenous infusion, significantly greater penetration of grepafloxacin was found in the iris, cornea and chorioretina of pigmented rabbits compared with albino rabbits.

Animals↗
PubMed2001

Study of retention in micellar liquid chromatography on a C8 column by the use of linear solvation energy relationships.

Linear solvation energy relationships (LSERs) are used to investigate the fundamental chemical interactions governing the micellar liquid chromatographic retention of 22 aromatic compounds (11 benzene derivatives and 11 aromatic polycyclic hydrocarbons) in 80 mobile phases on a C8 column. The systems studied involve combinations of 0.050 to 0.140 M sodium dodecyl sulfate or cetyltrimethylammonium bromide, with 0 to 10% methanol, n-propanol, and n-butanol as mobile phase modifiers. The ability of the LSERs to account for the chemical interactions underlying solute retention is shown. A comparison of predicted and experimental retention factors suggests that LSER formalism is able to reproduce adequately the experimental retention factors of the solutes studied in the different experimental conditions investigated.

Benzene Derivatives↗
PubMed2001

Analysis of Origanum vulgare volatiles by direct thermal desorption coupled to gas chromatography-mass spectrometry.

Volatile components of samples of a population of Spanish Origanum vulgare have been analyzed by direct thermal desorption coupled to GC-MS. The method is fast and reliable and requires a low amount of sample, allowing analysis of leaves and flowers from a single individual plant. Volatile yield is highly variable among individual plants and concentration also presents a high variation for most Origanum volatile compounds, linalool being the main component in most samples. Statistical analyses are applied in order to find patterns in composition data.

Adsorption↗
PubMed2001

The catalytic activity of dsRNA-dependent protein kinase, PKR, is required for NF-kappaB activation.

The double stranded RNA-dependent protein kinase (PKR), in addition to its role as a translational controlling factor, is a key transcriptional regulator exerting antiviral and antitumoral activities. We have previously shown that induction of NF-kappaB by PKR is involved in apoptosis commitment and this process is mediated through activation of the IKK complex. To gain insights into the mechanism of activation of NF-kappaB by PKR, we have analysed the domains of PKR involved in IKK activation and subsequent NF-kappaB induction. In PKR(0/0) cells infected with a collection of vaccinia virus (VV) recombinants expressing different mutant forms of PKR, we found that only PKR forms conserving the catalytic activity are able to activate NF-kappaB. An inactive PKR mutant (K296R), was unable to induce NF-kappaB activation despite full expression of the protein in a wide range of concentrations, as defined by Western blot, EMSA, IKK kinase activity and NF-kappaB transactivation assays. Moreover, the mutant PKR (K296R) acts as a dominant negative of PKR-induced eIF-2alpha phosphorylation and NF-kappaB activation. However, PKR mutants unable to activate NF-kappaB still retain their ability to associate with the IKK complex, as confirmed by immunoprecipitation analysis. We conclude that the catalytic activity of PKR and not only a protein-protein interaction with the IKK complex, is needed for activation of the transcription factor NF-kappaB.

Animals↗
PubMed2001

Composite starch-based coatings applied to strawberries (Fragaria ananassa).

Starch-based coatings were used to the extend storage life of strawberries (Fragaria ananassa) stored at 0 degree C and 84.8% relative humidity. Effects of coating formulation (including starch type, plasticizer, lipid and antimicrobial agent) were analysed with respect to fruit quality. Plasticizer addition was necessary for film and coating integrity to avoid pores and cracks. Plasticizer presence reduced weight losses and maintained surface colour of fruits. Amylomaize coatings showed lower water vapour and gas permeabilities and decreased weight losses for longer periods than corn starch ones. Coatings with sorbitol showed lower permeabilities than glycerol ones. Coatings with antimicrobial agents decreased microbial counts, extending storage life of coated fruits by 10 to 14 days in comparison to the control. The addition of 2 g/l sunflower oil to the formulations decreased the water vapour permeability of starch-based films, maintained the surface colour of coated fruits and controlled effectively fruit weight losses during storage. Lipid addition minimized the effects of starch and plasticizer types. Composite starch-based coatings showed selective gas permeability (CO2 higher than O2) which helps to delay senescence of fruits.

Biofilms↗
PubMed2001

Evolution and function of tandem repeats in the major surface protein 1a of the ehrlichial pathogen Anaplasma marginale.

The major surface protein (MSP) 1a of the ehrlichial cattle pathogen Anaplasma marginale, encoded by the single-copy gene msp1alpha, has been shown to have a neutralization-sensitive epitope and to be an adhesin for bovine erythrocytes and tick cells. msp1alpha has been found to be a stable genetic marker for the identification of geographic isolates of A. marginale throughout development in acutely and persistently infected cattle and in ticks. The molecular weight of MSP1a varies among geographic isolates of A. marginale because of a varying number of tandemly repeated peptides of 28-29 amino acids. Variation in the sequence of the tandem repeats occurs within and among isolates, and may have resulted from evolutionary pressures exerted by ligand-receptor and host-parasite interactions. These repeated sequences include markers for tick transmissibility that may be important in the identification of ehrlichial pathogens because they may influence control strategies and the design of subunit vaccines.

Amino Acid Sequence↗
PubMed2000

Identification of amino acid residues of transcription factor AP-2 involved in DNA binding.

AP-2 is a cell-type specific, developmentally regulated transcription factor which has been described as a critical regulator of gene expression during vertebrate development and embryogenesis. Although the overall domains of this factor necessary for their activity have been identified, the exact identity of AP-2 amino acid residues responsible for its interaction with the DNA structure has not yet been described. Here, we describe the identification of a region of AP-2 which was protected by an oligonucleotide probe containing its binding site from trypsin digestion, monitored by peptide mapping by MALDI-TOF mass spectrometry. Furthermore, we analyzed the relative in vitro DNA-binding activity, the stimulatory potency on the AP-2-dependent APOE promoter, as well as the ability to inhibit the effect of the wild-type protein of each one of a set of single-site substitution AP-2 mutants spanning the identified region. Taken together, our data clearly demonstrate that the region between amino acid residues 252-260 of AP-2 is essential for its DNA-binding activity. Particularly, the individual substitution in any of the residues 253, 254, 255, 257 or 260 is sufficient for completely abolishing the interaction with DNA and the stimulation of APOE promoter activity. These results indicate a crucial role of this region in the formation of an active DNA-binding domain and strongly suggest that these residues provide direct contacts with the DNA structure at the AP-2 binding site.

Amino Acid Sequence↗
PubMed2000

Separation of a group of N-phenylpyrazole derivatives by micellar electrokinetic chromatography: application to the determination of solute-micelle association constants and estimation of the hydrophobicity.

Micellar electrokinetic chromatography (MEKC) was applied to the separation of a group of N-phenylpyrazole derivatives. Sodium dodecyl sulfate (SDS) as micellar system and 2-(N-cyclohexylamino)ethanesulfonic acid (CHES) as separation buffer (pH 10) were employed in the absence and presence of different percentages of medium chain alcohols (n-propanol or n-butanol). The separation of multicomponent mixtures of the solutes studied enabled the rapid determination of their retention factors which, in turn, allowed the study of the separation selectivity of compounds and the determination of their solute-micelle association constants (from the linear variation of the retention factors as a function of the total surfactant concentration in the separation buffer). Separation selectivity was studied according to the elution range and number of solutes separated in all the electrolyte solutions employed (45 micellar phases). The effect of the buffer concentration (0.05, 0.08 and 0.10 M), the alcohol nature (n-propanol or n-butanol) and the alcohol percentage (1, 3 or 5%) of the values obtained for the solute-micelle association constants was also studied. The best separation (12 solutes) was performed when a 0.08 M CHES buffer, pH 10, 0.02 M SDS modified by 5% n-butanol was used. The possibilities of using MEKC for evaluating the hydrophobicity of compounds was investigated through the study of the correlation between the logarithm of the retention factors of N-phenylpyrazole derivatives and their logarithm of the octanol-water distribution coefficients estimated by high performance liquid chromatography (HPLC).

Alcohols↗
PubMed2000

Accumulation of mercury in edible macrofungi: influence of some factors.

The levels of mercury in terrestrial ecosystems are generally very low, but the strong accumulation in some species of macrofungi is an exception from this rule. Mercury contents of 41 samples of edible mushrooms fruiting bodies representing eight species (six wild and two cultivated) were determined by an anodic stripping voltammetric technique (ASV) using a gold disc as the working electrode. Wild fungi were collected in unpolluted and polluted areas in the Province of Lugo (NW Spain). Influence of some factors (ecology, species, traffic pollution, and morphological portion) and the importance of mushrooms as a dietary source of mercury have been studied. Wild saprophytic species showed higher levels (ppm DW) than mycorrhizals species, with the exception of Boletus pinicola. There were significant differences according to the species analyzed (p < 0.001), the highest average content of mercury was found in Boletus pinicola (7.37 ppm DW), and the range was 0.35-33.07 ppm DW for hymenophore and 0.18-20.30 ppm DW for the rest of the fruit body. The cultivated species accumulated lower than wild species because the mean life is shorter. The traffic pollution factor did not show significant differences, so mushrooms are not realiable bioindicators of traffic pollution by mercury. Hymenophore was always the morphological portion that contained the highest mercury levels (p < 0.05), and the mean ratio of hymenophore/rest of the fruit body was 2.13. The mercury concentrations were compared to literature data and levels set by legislation, and the contribution of mushrooms to the weekly intake of mercury per person was evaluated. The possible health risk for people is pointed out.

Agaricales↗
PubMed2000

Organochlorine and heavy metal residues in the water/sediment system of the Southeast Regional Park in Madrid, Spain.

A study into levels of contamination by organochlorine compounds (insecticides and PCBs) and heavy metals (Cd and Pb) in the water/sediment system of the Southeast Regional Park (SERP) in Madrid, Spain, has been carried out. Residue levels of xenobiotics were determined in surface and underground waters and sediments from selected sites throughout the protected area. The results showed these contaminants to be widespread throughout the studied area. p,p'-DDT concentration levels were consistently higher than its metabolite p,p'-DDE, indicating a recent use of this organochlorine insecticide in the area. PCB levels exceeded, in the majority of the cases, the levels taken as the maximum (100 ng/microl) for highly polluted waters. Cd and Pb levels found in water samples were under the detection limits of the methodology used. Pb levels found in sediment samples were higher than Cd.

Benzophenones↗
PubMed2000

[Client categorization: a tool to assess nursing workload].

BACKGROUND: Client categorization is a management tool that allows an objective and structured assessment of the care demands imposed by patients and nursing workloads. AIM: To characterize the need for direct nursing care of patients admitted to a university hospital. PATIENTS AND METHODS: During two months, all patients admitted to intensive, intermediate care units and general services were categorized, their need for nursing care and the time invested by nurses in their care was registered. All patients were classified as maximal, high, median or low risk and as independent, totally or partially dependent on nursing care. Considering four degrees of risk and three degrees of dependency, 12 categories of patients were defined. RESULTS: Patients admitted to intensive care units were of maximal risk and totally dependent and required 1 nurse per 2.2 patients. Those admitted in intermediate care units required 1 nurse per 3.8 patients and those in general services, 1 nurse per 11.5 patients. CONCLUSIONS: Client categorization is a reproducible method that determines a standard measuring unit to define nursing needs. This allows the comparison of workloads between different services within a hospital or between hospitals.

Humans↗
PubMed1999

Transcription factor AP-2 activity is modulated by protein kinase A-mediated phosphorylation.

We recently reported that APOE promoter activity is stimulated by cAMP, this effect being mediated by factor AP-2 [Garcia et al. (1996) J. Neurosci. 16, 7550-7556]. Here, we study whether cAMP-induced phosphorylation modulates the activity of AP-2. Recombinant AP-2 was phosphorylated in vitro by protein kinase A (PKA) at Ser239. Mutation of Ser239 to Ala abolished in vitro phosphorylation of AP-2 by PKA, but not the DNA binding activity of AP-2. Cotransfection studies showed that PKA stimulated the effect of AP-2 on the APOE promoter, but not that of the S239A mutant. Therefore, cAMP may modulate AP-2 activity by PKA-induced phosphorylation of this factor.

Amino Acid Substitution↗
PubMed1999

High-sensitivity analysis and sequencing of peptides and proteins by quadrupole ion trap mass spectrometry.

This paper describes experience with the commercially available LCQ quadrupole ion trap mass spectrometer applied to the off-line analysis of peptides and proteins. The standard front end of the electrospray probe was replaced with a micromanipulator which, with the aid of a magnifying device, allowed the use of a variety of miniaturized spraying interfaces. The low sample consumption and extended analysis times of these devices were ideally suitable to obtain improved results in terms of sensitivity and mass accuracy. This needed a careful optimization of the number of ions stored inside the trap (ion target parameter) and required spectrum averaging of many scans. A method is presented for the mathematical fitting of ZoomScan spectra to theoretical isotopic distributions, which allowed the mass determination of large peptides with more accuracy than that achieved by conventional deconvolution algorithms. A very simple on-line desalting configuration is also described which needed no external micro-high-performance liquid chromatographic pumps, and can be easily mounted using the built-in syringe delivery system of the LCQ. This set-up allowed extended analysis times of 'in-gel' protein digests in subpicomole amounts. Finally, the multiple fragmentation capabilities of the ion trap were found to be extremely useful for the analysis of peptide modifications such as phosphorylation and for sequencing individual peptides from highly complex MHC-bound peptide pools.

Animals↗