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Biomedical subjects

P García

Publications and source records attributed to P García.

At least 55 records · Page 3Linked to original sources
PubMed1999

A2 cro, the lysogenic cycle repressor, specifically binds to the genetic switch region of Lactobacillus casei bacteriophage A2.

Lysogenic induction of temperate bacteriophage A2 of Lactobacillus casei is controlled by the action of its cI and cro products at the phage operator region. Three 20-bp inverted repeated DNA segments (subsites O1, O2, and O3) and the two divergent (PL and PR) promoters were mapped within the 153-bp operator region. The A2-encoded Cro product is shown to be the functional homolog of lambda Cro. The binding of Cro to the three operator subsites is noncooperative and yields two discrete protein-DNA complexes of retarded migration in mobility shift assays. The Kapp value for the Cro-PL-PR DNA complex was estimated to be 6 nM. Cro shows a slightly higher affinity for O3 than for O1 and O2 subsites. The O3 subsite overlaps the -35 hexamer of the PL promoter, which directs cI expression. A Cro mutant protein, devoid of the last 12 residues (Cro*), allowed the assignment of the DNA-binding domain to the NH2 end of Cro. The C end enhances its affinity for the DNA and probably stabilizes bending induced by Cro.

Base Sequence↗
PubMed1999

Spanish population data on the four STR loci D8S1179, D16S539, D18S51 and D21S11.

Population data were generated for four tetrameric short tandem repeat loci systems (D8S1179, D16S539, D18S51 and D21S11) for a Spanish Caucasian population sample (n = 218-219 individuals) using PCR. All loci were highly polymorphic, met Hardy-Weinberg expectations and the results demonstrated the assumption of independence of the loci analysed. The allele frequency data can be used in identity testing to estimate the frequency of a multiple PCR-based DNA profile in the Spanish population.

Alleles↗
PubMed1999

Preliminary study on genetic variability of Dicrocoelium dendriticum determined by random amplified polymorphic DNA.

Genetic variability of adult specimens of Dicrocoelium dendriticum has been studied using random amplified polymorphic DNA (RAPD). The worms were collected from the infected livers of different sheep from several localities in León province (NW Spain). DNA fragments were amplified by means of decamer primer oligonucleotides of arbitrary sequence. Some primers produce complex and highly variable patterns of amplified DNA in D. dendriticum. Intra- and inter-population genetic variability of adult parasites were analyzed, scoring polymorphic and monomorphic reproducible bands by means of the Jaccard similarity, and dendrograms showing genetic relationships between individuals were obtained using the FITCH method. Genetic variability seems to be high in this parasite and genetic similarity within a population (worms infecting a single animal) is similar to the average similarity between worms from different sheep. These results suggest that each sheep is infected by numerous genetically different parasites from one or more populations of infected ants.

Animals↗
PubMed1999

Phenolic compounds in Spanish olive oils.

Phenolic compounds in Spanish virgin olive oils were characterized by HPLC. Simple phenols such as hydroxytyrosol, tyrosol, vanillic acid, p-coumaric acid, ferulic acid, and vanillin were found in most of the oils. The flavonoids apigenin and luteolin were also found in most of the oils. The dialdehydic form of elenolic acid linked to tyrosol and hydroxytyrosol was also detected, as were oleuropein and ligstroside aglycons. The structure of a new compound was elucidated by MS and NMR as being that of 4-(acetoxyethyl)-1,2-dihydroxybenzene. Changes of phenolic compounds in virgin olive oils with maturation of fruits were also studied. Hydroxytyrosol, tyrosol, and luteolin increased their concentration in oils with maturation of fruits. On the contrary, glucoside aglycons diminished their concentration with maturation. No clear tendency was observed for the rest of the phenolic compounds identified.

Chromatography, High Pressure Liquid↗
PubMed1999

The molecular characterization of the first autolytic lysozyme of Streptococcus pneumoniae reveals evolutionary mobile domains.

A biochemical approach to identify proteins with high affinity for choline-containing pneumococcal cell walls has allowed the localization, cloning and sequencing of a gene (lytC ) coding for a protein that degrades the cell walls of Streptococcus pneumoniae. The lytC gene is 1506 bp long and encodes a protein (LytC) of 501 amino acid residues with a predicted M r of 58 682. LytC has a cleavable signal peptide, as demonstrated when the mature protein (about 55 kDa) was purified from S. pneumoniae. Biochemical analyses of the pure, mature protein proved that LytC is a lysozyme. Combined cell fractionation and Western blot analysis showed that the unprocessed, primary product of the lytC gene is located in the pneumococcal cytoplasm whereas the processed, active form of LytC is tightly bound to the cell envelope. In vivo experiments demonstrated that this lysozyme behaves as a pneumococcal autolytic enzyme at 30 degrees C. The DNA region encoding the 253 C-terminal amino acid residues of LytC has been cloned and expressed in Escherichia coli. The truncated protein exhibits a low, but significant, choline-independent lysozyme activity, which suggests that this polypeptide adopts an active conformation. Self-alignment of the N-terminal part of the deduced amino acid sequence of LytC revealed the presence of 11 repeated motifs. These results strongly suggest that the lysozyme reported here has changed the general building plan characteristic of the choline-binding proteins of S. pneumoniae and its bacteriophages, i.e. the choline-binding domain and the catalytic domain are located, respectively, at the N-terminal and the C-terminal moieties of LytC. This work illustrates the natural versatility exhibited by the pneumococcal genes coding for choline-binding proteins to fuse separated catalytic and substrate-binding domains and create new and functional mature proteins.

Amino Acid Sequence↗
PubMed1999

Effect of four enteral foods on the small bowel of undernourished rats after midgut resection.

OBJECTIVE: To describe the effects of new enteral foods on the adaptation of the gut mucosa after massive intestinal resection in rats. DESIGN: Laboratory experiment. SETTING: Teaching hospital, Spain. SUBJECTS: 91 male Wistar rats, 69 of which were studied (5 were excluded and 17 died). INTERVENTIONS: Previously undernourished rats were subjected to either massive bowel resection (n = 30) or laparotomy (n = 26) and fed four enteral hypocaloric diets for 7 days: Alitraq (n = 7 in each group), Impact (n = 8 and 7), Enrich (n = 8 and 6), and Elemental (n = 7 and 6). The remainder were not operated on and fed chow (n = 7) or a diet containing no protein (n = 6). Two diets were high in protein (Alitraq and Impact) and two contained normal amounts (Enrich and Elemental). MAIN OUTCOME MEASURES: Bowel mucosal thickness and proliferation; disaccharidase activity; intestinal weight and length; body weight; and plasma somatostatin, IGF-1, and peptide YY concentrations. RESULTS: Enriched diets provided a higher body and intestinal weight, and increased length of jejunal and ileal villous size. Peptide concentrations were modified by resection but not by the diet given. Concentrations of somatostatin and insulin-like growth factor were reduced in all groups with the exception of somatostatin in the two diets high in protein in the sham-operated rats. CONCLUSIONS: Enriched diets all improve the intestinal adaptive response to massive bowel resection in rats, offering advantages over diets with normal amounts of protein.

Adaptation, Physiological↗
PubMed1999

Cooperative interaction of CI protein regulates lysogeny of Lactobacillus casei by bacteriophage A2.

The temperate bacteriophage A2 forms stable lysogens in Lactobacillus casei. The A2-encoded cI product (CI), which is responsible for maintaining the A2 prophage in the lysogenic state, has been purified. The CI protein, which is a monomer of 25.3 kDa in solution, specifically binds to a 153-bp DNA fragment that contains two divergent promoters, PL and PR. These promoters mediate transcription from cI and a putative cro, respectively. Three similar, although not identical, 20-bp inverted repeated DNA segments (operator sites O1, O2, and O3) were found in this segment. CI selectively interacts with O1, which is placed downstream from the transcription start point of the cro gene, and with O2 and O3, which overlap with the -35 region of the two promoters. Using a heterologous RNA polymerase, we have determined the transcription start points of PL and PR. CI exerts a negative effect on the in vitro transcription of PR by repositioning the RNA polymerase in a concentration-dependent manner. CI, when bound to O1 and O2, enhances the positioning of the RNA polymerase with the PL promoter. Our data indicate that the CI protein regulates the lytic and lysogenic pathways of the A2 phage.

Bacteriophages↗
PubMed1998

Immunohistochemical localization of epidermal growth factor, transforming growth factor-alpha and growth factor-beta s in the caprine peri-implantation period.

Control over the action of steroid hormones in the uterus and conceptus during the initial period of gestation appears to be regulated locally by growth factors. This study involved immunohistochemical detection of epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha) and transforming growth factor-beta s (TGF-beta s), to determine their role in the caprine peri-implantation period. Epidermal growth factor was expressed in the luminal and glandular endometrial epithelium of goats on all days studied (Days 22 to 30 post coitum), but it was not detected in trophoblastic cells or in other embryonic structures. Between Days 22 and 30 post coitum, TGF-alpha was detected in the epithelial cells and superficial stroma of the uterus and in the trophoendodermic cells of the embryo. Transforming growth factor-beta s expression, observed in the endometrium, embryo and extraembryonic membranes on Day 22 post coitum, decreased by Day 24 post coitum and disappeared in the embryo by Day 30 post coitum, while remaining in the other structures. The presence of these growth factors during the peri-implantation period in the goat suggests their participation in proliferation and differentiation phenomena which occur during implantation and embryonic development.

Animals↗
PubMed1998

Unsupervised classification of neural spikes with a hybrid multilayer artificial neural network.

The understanding of the brain structure and function and its computational style is one of the biggest challenges both in Neuroscience and Neural Computation. In order to reach this and to test the predictions of neural network modeling, it is necessary to observe the activity of neural populations. In this paper we propose a hybrid modular computational system for the spike classification of multiunits recordings. It works with no knowledge about the waveform, and it consists of two moduli: a Preprocessing (Segmentation) module, which performs the detection and centering of spike vectors using programmed computation; and a Processing (Classification) module, which implements the general approach of neural classification: feature extraction, clustering and discrimination, by means of a hybrid unsupervised multilayer artificial neural network (HUMANN). The operations of this artificial neural network on the spike vectors are: (i) compression with a Sanger Layer from 70 points vector to five principal component vector; (ii) their waveform is analyzed by a Kohonen layer; (iii) the electrical noise and overlapping spikes are rejected by a previously unreported artificial neural network named Tolerance layer; and (iv) finally the spikes are labeled into spike classes by a Labeling layer. Each layer of the system has a specific unsupervised learning rule that progressively modifies itself until the performance of the layer has been automatically optimized. The procedure showed a high sensitivity and specificity also when working with signals containing four spike types.

Animals↗
PubMed1998

HLA-E-bound peptides influence recognition by inhibitory and triggering CD94/NKG2 receptors: preferential response to an HLA-G-derived nonamer.

The HLA-E class Ib molecule constitutes a major ligand for the lectin-like CD94/NKG2 natural killer (NK) cell receptors. Specific HLA class I leader sequence-derived nonapeptides bind to endogenous HLA-E molecules in the HLA-defective cell line 721.221, inducing HLA-E surface expression, and promote CD94/NKG2A-mediated recognition. We compared the ability of NK clones which expressed either inhibitory or activating CD94/NKG2 receptors to recognize HLA-E molecules on the surface of 721.221 cells loaded with a panel of synthetic nonamers derived from the leader sequences of most HLA class I molecules. Our results support the notion that the primary structure of the HLA-E-bound peptides influences CD94/ NKG2-mediated recognition, beyond their ability to stabilize surface HLA-E. Further, CD94/ NKG2A+ NK clones appeared more sensitive to the interaction with most HLA-E-peptide complexes than did effector cells expressing the activating CD94/NKG2C receptor. However, a significant exception to this pattern was HLA-E loaded with the HLA-G-derived nonamer. This complex triggered cytotoxicity very efficiently over a wide range of peptide concentrations, suggesting that the HLA-E/G-nonamer complex interacts with the CD94/NKG2 triggering receptor with a significantly higher affinity. These results raise the possibility that CD94/NKG2-mediated recognition of HLA-E expressed on extravillous cytotrophoblasts plays an important role in maternal-fetal cellular interactions.

Antigens, CD↗
PubMed1998

Growth hormone secretory response to intravenous galanin infusion in acromegalic patients.

Galanin is a 29-amino acid neuropeptide which stimulates the secretion of growth hormone (GH) in normal men. Although the diagnosis of acromegaly involves demonstrating hypersecretion of GH and/or abnormalities in GH secretory dynamics, sometimes it is difficult to establish the activity of the disease. The aim of our study was to assess the response to galanin infusion in acromegalic patients (active and cured). We studied 19 subjects: 5 healthy volunteers (group I), 9 patients with active acromegaly (group II), and 5 with acromegaly cured after transsphenoidal surgery (group III). We performed a test of infusion with porcine galanin (8 microg/Kg/h) to study the secretory response of the GH. Galanin produced a marked increase in GH in the controls, group I (F9,36 = 5.34; p < 0.01) and in the cured patients, group III (F9,36 = 7.35; p < 0.01). On the other hand, galanin did not significantly modify the secretion of GH in the patients with active disease, group II. The areas under the curve (AUC) were higher in groups I and III compared to group II (p < 0.01). Nevertheless, there were no statistically significant differences in the AUC of groups I and III. Our results indicate that the differences in the GH response to galanin in patients with active and cured acromegaly could be of value in the study of the disease.

Acromegaly↗
PubMed1998

Growth hormone reduces mortality and bacterial translocation in irradiated rats.

Growth hormone stimulates the growth of intestinal mucosa and may reduce the severity of injury caused by radiation. Male Wistar rats underwent abdominal irradiation (12 Gy) and were treated with either human growth hormone (hGH) or saline, and sacrificed at day 4 or 7 post-irradiation. Bacterial translocation, and the ileal mucosal thickness, proliferation, and disaccharidase activity were assessed. Mortality was 65% in irradiated animals, whereas hGH caused a decrement (29%, p < 0.05). Bacterial translocation was also reduced by hGH (p < 0.05). Treating irradiated rats with hGH prevented body weight loss (p < 0.05). Mucosal thickness increased faster in irradiated hGH-treated animals. The proliferative index showed an increment in hGH-treated animals (p < 0.05). Giving hGH to irradiated rats prevented decrease in sucrose activity, and increment in lactase activity. In conclusion, giving hGH to irradiated rats promotes the adaptative process of the intestine and acute radiation-related negative effects, including mortality, bacterial translocation, and weight loss.

Abdomen↗
PubMed1998

Histological and immunohistochemical study of the cat epididymis.

A histological and immunohistochemical study of the epididymis of 21 cats which ranged from 7 to 8 months to 2 years of age was made. Five different cell types were observed in the feline epididymis: principal and basal cells, which were the most numerous, apical and narrow, PAS positive cells, which were scarce, and migratory cells, consisting of lymphocytes and macrophages. Four morphologically distinct areas of the cat epididymis were identified. Region I displayed a stellate lumen and the principal cells were tall and presented long and irregular stereocilia. In region II, the lumen was oval and the principal cells were shorter than in the initial area. Region III displayed characteristics similar to those of region I; the principal difference was the presence of short and regular stereocilia on the surface of the principal cells. In region IV the lumen was very spacious and the epithelium shorter than in the other regions. In regions I, II, and III intraepithelial cavities were observed. With regard to the immunohistochemical results, the basal cells displayed medium immunoreactivity with vimentin. Due to its anti-desmin reactivity, the muscle wall which surrounds the epididymis was seen to become progressively thicker as it nears the tail. Broad-spectrum anti-cytokeratin serum produced intense immunostaining in the basal, apical and narrow, PAS positive cells; the principal cells of region IV also displayed a strong immunoreactivity.

Aging↗
PubMed1998

Identification of the repressor-encoding gene of the Lactobacillus bacteriophage A2.

The repressor gene of the Lactobacillus phage A2 has the following properties: it (i) encodes a 224-residue polypeptide with DNA binding and RecA cleavage motifs, (ii) is expressed in lysogenic cultures, and (iii) confers superinfection immunity on the host. Adjacent, but divergently transcribed, lies another open reading frame whose product resembles the lambda Cro protein. In the 161-bp intergenic segment, putative promoters and operators have been detected.

Amino Acid Sequence↗