Endocarditis infecciosa en enfermedad por mordedura de rata / Infectious endocarditis in rat bite disease

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Sección de los músculos rectos delabdomen por cinturón de seguridad / Section of rectus abdominis muscle by the safety belt

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Cloning and molecular characterization of the acyl-CoA: diacylglycerol acyltransferase 1 (DGAT1) gene from Echium.

Boraginaceae species, such as those from the genus Echium, contain high levels of the Delta(6)-desaturated gamma-linolenic (18:3n-6) and octadecatetraenoic (18:4n-3) acids. These are unusual fatty acids among the plant kingdom that are gaining interest due to their benefits to human health. The potential utility of acyltransferases aimed at an increase in oil yield and fatty acid profiling has been reported. In this work, a gene encoding an acyl-CoA:diacylglycerol acyltransferase (DGAT, EC 2.3.1.20) was cloned from Echium pitardii. Genomic and cDNA sequences obtained revealed a gene structure composed of 16 exons, yielding a protein (EpDGAT) of 473 amino acids with high similarity to DGAT1 enzymes of plants. Protein features such as a predicted structure with a highly hydrophilic N-terminus followed by 10 transmembrane domains, as well as the presence of diverse specific signatures, also indicate that EpDGAT belongs to the DGAT1 family. indeed. DGAT activity of the protein encoded by EpDGAT was confirmed by heterologous expression of the full-length cDNA in a yeast mutant (H1246) defective in the synthesis of triacylglycerols. Fatty acid composition of the triacylglycerols synthesized by EpDGAT in H1246 yeast cultures supplemented with polyunsaturated fatty acids suggest a substrate preference for the trienoic fatty acids alpha-linolenic acid (18:3n-3) and gamma-linolenic acid over the dienoic linoleic acid (18:2n-6). Site-directed mutagenesis has revealed the presence of a critical residue (P(178) in EpDGAT) within a reported thiolase signature for binding of acyl-enzyme intermediates that might be involved in the active site of the enzyme. Transcript analysis for EpDGAT shows an ubiquitous expression of the gene which is increased in leaves during senescence.

Estancias inadecuadas en el Servicio de Medicina Interna de un Hospital Comarcal. AEP como herramienta útil en la gestión / lnadequate hospital rooms in the Interna Medicine Service of a Regional Hospital. AEP as a useful management tool

El objetivo de este trabajo es estudiar la inadecuación de estancias hospitalarias en el Servicio de Medicina Interna del Hospital Infanta Margarita de Cabra. Para la consecución de este objetivo, se utilizó la herramienta AEP (The Appropriateness Evaluation Protocol). El estudio nos desveló una inadecuación del 21, 7% de las estancias. Los resultados obtenidos se encuentran dentro del margen reflejado en otros estudios, que sitúan la inadecuación entre el 20-40%. Se puede concluir que los resultados obtenidos son adecuados, además de que nos ha permitido detectar oportunidades de mejora en el manejo de los pacientes. (AU)

Study of the inadequacy of hospital rooms in the Internal Medicine Service of the Hospital Infanta Margarita de Cabra which has led to detecting opportunities far improvement in the handling of patients (AU)

Estudio epidemiológico de las parasitosis intestinales 2000-2001 en población ambulatoria del Área 1 de la Comunidad de Madrid / Epidemiological study of intestinal parisitism 2000-2001 of outpatients in a health area (Madrid, Spain)

Objetivo: Conocer la prevalencia de parasitosis intestinales en población ambulatoria de nuestra Área Sanitaria de Salud. Diseño: Estudio retrospectivo observacional. Marco poblacional Área Sanitaria 1 de la Comunidad de Madrid, España. Material y métodos: Durante el período de estudio (enero 2000-diciembre 2001) han sido procesadas 6.137 muestras fecales (5.783 heces y 354 muestras para diagnóstico de oxiuros, 312 torundas vaselinizadas y 42 celos) procedentes de 3.396 pacientes. Las heces fueron procesadas mediante técnica de concentración en formol-acetato de etilo. La detección de ooquistes de coccidios se llevó a cabo mediante tinción de ácido-alcohol resistencia de Kinyoun modificada, de frotis directos y concentrados. Resultados: La prevalencia de parasitación intestinal ha sido del 13,5 per cent (827 muestras fecales positivas, pertenecientes a 464 pacientes, detectándose un total de 1.029 parásitos). La distribución de los parásitos observados y su prevalencia ha sido: Blastocystis hominis (39,3 per cent), Giardia intestinalis (19,3 per cent), Entamoeba coli y Endolimax nana (12,0 per cent cada uno), Enterobius vermicularis (6,9 per cent), Cryptosporidium parvum (3,6 per cent), Entamoeba histolytica/dispar (3,2 per cent), Trichuris trichiura (1,8 per cent), Hymenolepis nana (0,8 per cent), Taenia saginata (0,5 per cent), Ascaris lumbricoides (0,4 per cent) y Uncinarias (0,1 per cent). Los parásitos intestinales identificados más frecuentemente han sido: Giardia intestinalis (52,8 per cent), Enterobius vermicularis (18,8 per cent), Cryptosporidium parvum (9,8 per cent) y Entamoeba histolytica/dispar (8,7 per cent). Han sido analizados el origen de las muestras, edad, distribución por sexos y procedencia de los pacientes así como la incidencia estacional de los parásitos intestinales detectados. Conclusiones: Nuestros hallazgos sugieren que las infecciones parasitarias todavía constituyen un problema de salud pública. El conocimiento de la situación en cada área facilita su manejo y control (AU)

Occurrence and characterization of oils rich in gamma-linolenic acid (III): the taxonomical value of the fatty acids in Echium (Boraginaceae).

Fourteen species of the genus Echium (Fam. Boraginaceae) collected in the Macaronesia were surveyed in a search for high levels of gamma-linolenic acid (GLA, 18:3omega6) in the seed oil. High amounts of this fatty acid were found in all of them, ranging from 18.85% (E. pitardii var. pitardii) to 27.42% (E. gentianoides) on total seed fatty acids. The GLA content related to total seed weight was also significant, ranging from 1.26% (E. handiense) to 8.22% (E. gentianoides). In addition, considerable amounts of stearidonic acid (SA, 18:4omega3) were detected, ranging from 3.78% (E. bonnetii var. bonnetii) to 8.81% (E. pininana) on total fatty acids. Besides all the perennial species, the four herbaceous Echium taxa endemic to the Macaronesia also showed high GLA percentages. This is in contrast to the low GLA level found in continental Echium species, all of them bearing an herbaceous habit. These results are in good agreement with the available genetic data and show the ability of GLA to discriminate between Macaronesian and continental Echium species. The analysis of five other Macaronesian species belonging to plant families rich in GLA are also reported.

Genomic organization and transcriptional analysis of STDEFICIENS in Solanum tuberosum L.

The genomic organization of STDEFICIENS (STDEF), the potato orthologous gene to DEFICIENS (DEF) from Antirrhinum majus and APETALA3 (AP3) from Arabidopsis thaliana, has been investigated. Southern-blot analysis on genomic DNA from dihaploid potato lines, using 5'-gene specific probes, revealed polymorphisms that were consistent with the existence in potato of at least two copies of STDEF per haploid genome. This was confirmed by the detection of at least six different STDEF transcripts in the common tetraploid potato S. tuberosum. Genes for two of the STDEF loci, here designated as STDEF-1 and STDEF-2, have been identified as corresponding to the previously described pD13 and pD12 genomic clones, respectively (García-Maroto et al., 1993). In addition we have characterised the transcriptional STDEF unit. The main transcription start has been identified around 90 nt upstream of the putative initiation ATG codon, at a CAAATC motif, conserved in AP3. An additional transcription initiation site was detected by 5'-RACE analysis about 300 nt upstream of the main start, which has been confirmed by reverse transcriptase - polymerase chain reaction (RT-PCR) amplification from the longer transcripts. A comparison of the promoter regions for pD12, pD13 and AP3 indicates a similar overall structure, but reveals the existence of a great divergence between pD12 and pD13 in a promoter region that should contain important cis-regulatory elements. This raises the possibility of a differential regulation for the two STDEF genes.

Occurrence and characterization of oils rich in gamma-linolenic acid part II: fatty acids and squalene from Macaronesian Echium leaves.

Leaves from 25 Macaronesian Echium (Boraginaceae) species have been surveyed for hydrocarbon compounds. These plants were previously reported as the major source of gamma-linolenic acid so far found in nature. In addition, six European Echium species and the common Borago officinalis have been analysed for comparative purposes. High squalene amounts were found in all Echium plants from the Macaronesia, ranging from 3.73%, in E. simplex to 20.1% in E. fastousum. Squalene was almost absent from all European Echium species, and the same is true for B. officinalis. The relatively high oil content (2.27%) in leaves of E. fastuosum raises the total squalene amount to about 0.46% within this tissue. The main fatty acid component in the leaf was alpha-linolenic acid (ALA, 18:3omega3), ranging in the Macaronesian Echium from 9.32% in E. acanthocarpum to 54.45% in E. simplex. Possible utilisation of these plants as a commercial source of squalene and hypotheses about its physiological role in the plant are discussed.

Cloning, mapping and expression analysis of barley MADS-box genes.

Six MADS-box cDNA clones were isolated by heterologous screening from a barley inflorescence cDNA library. Based on sequence comparison to known MADS-box genes, the barley MADS-box (BM) genes were grouped into three distinct phylogenetic subclasses of the MADS-box gene family. The three MADS-box genes BM3, BM5 and BM8 share similarities with genes of the SQUAMOSA (SQUA) subgroup, while BM7 and BM9 belong to the AGAMOUS-LIKE 2 (AGL2) subgroup. BM1 resembles MADS-box genes described as solitary sequences or orphan genes. Expression analysis of the barley MADS-box genes revealed expression patterns that are not characteristic of the barley MADS-box genes of the SQUA subgroup. while expression of BM7 and BM9 was largely as expected for the AGL2 subgroup. BM1 is mainly expressed in vegetative tissues and its primary transcript undergoes alternative splicing such that the corresponding mRNAs differ by two codons. The genes BM1, BM3 and BM8 were mapped by analysis of single-nucleotide polymorphisms onto barley chromosomes 4, 2 and 7, respectively.

Characterization of the potato MADS-box gene STMADS16 and expression analysis in tobacco transgenic plants.

A new MADS-box gene, STMADS16, has been cloned in Solanum tuberosum L. that is expressed in all vegetative tissues of the plant, mainly in the stem, but not in flower organs. STMADS16 expression is established early during vegetative development and is not regulated by light. Sequence similarity besides the spatial and temporal expression patterns allow to define a novel MADS-box subfamily comprising STMADS16 and the gene STMADS11. Expression of the STMADS16 sense cDNA under the control of the 35S cauliflower mosaic virus promoter modifies the inflorescence structure by increasing both internode length and flower proliferation of the inflorescence meristems, and confers vegetative features to the flower. Moreover, STMADS16 ectopic expression overcomes the increase in flowering time and node number produced under short-day photoperiod, while the flowering time is not affected in long-day conditions. These results are discussed in terms of a possible role for STMADS16 in promoting vegetative development.

Occurrence and characterization of oils rich in gamma-linolenic acid Part I: Echium seeds from Macaronesia.

Nineteen species of the genus Echium (Fam. Boraginaceae) collected in Macaronesia were surveyed in a search for new sources of gamma-linolenic acid (GLA, 18:3omega6). High amounts of this acid were found in all of them, ranging from 9.15% (E. plantagineum) to 26.31% (E. callithyrsum) of total seed fatty acids. The amounts of GLA related to total seed weight were also significant, ranging from 1.77% (E. sventenii) to 5.02% (E. nervosum). In addition, considerable amounts of stearidonic acid (SA, 18:4omega3) were detected, ranging from 3.03% (E. auberianum) to 12.94% (E. plantagineum) of total fatty acids. These data allow us to consider tile members of the genus Echium from Macaronesia as one of the richest sources of gamma-linolenic acid found so far in nature. The results obtained from multivariable data analysis and the taxonomic relationships among the species is discussed.

Plants as 'chemical factories' for the production of polyunsaturated fatty acids.

Polyunsaturated fatty acids (PUFAs) are valuable products because of their involvement in several aspects of human health. Market demand for most PUFAs is growing continually and current sources are considered insufficient for satisfying this demand; alternative sources are actively sought after. Oilseed plants can be a potential source of PUFAs if they are appropriately gene engineered. Most of the basic tools for genetic engineering of oilseed plants for giving them the ability to produce PUFAs are already developed. Here we review the prospects of genetic engineering of oilseed plants for producing some valuable long-chain polyunsaturated fatty acids. Genetic transformation for GLA production seems to be a near-term possibility, but gene engineering seems considerably more difficult for the other long-chain PUFAs. Nevertheless, with the current rapid pace of biotechnological advancement, the remaining difficulties may be surmounted in the near future.

Isolation and molecular characterization of a new vegetative MADS-box gene from Solanum tuberosum L.

A cDNA clone, STMADS11, encoding a new MADS-box protein was isolated from Solanum tuberosum L. (potato). Expression of STMADS11 was found in all vegetative organs of the plant, but not in floral tissues. The expression was also detected in all developmental stages, from tuber sprouts to mature plants, reaching a maximum in well-developed organs. However, the level of STMADS11 mRNA was low in tissues such as resting tuber or sprouts developed in the cold, where the metabolic activity is reduced. "In situ" hybridizations performed on leaf and stem sections showed that the STMADS11 transcript is mainly associated with vascular bundles. Cladistic analysis arising from amino acid sequence comparison revealed that STMADS11 shows the highest similarity to STMADS16, another vegetative MADS-box gene from potato, and to the previously reported "orphan" genes AGL15 and AGL17 from Arabidopsis thaliana. Possible implications of these data in relation to STMADS11 function are discussed.

The barley Hooded mutation caused by a duplication in a homeobox gene intron.

In barley (Hordeum vulgare L.) the unit of inflorescence is the spikelet, which bears a fertile bract, the lemma, and the floret consisting of palea, two lodicules, three stamens and the pistil. The Hooded mutation causes the appearance of an extra flower of inverse polarity on the lemma. This phenotype is governed by the single dominant genetic locus K3. Here we show that the homeobox gene Knox3 represents this locus. Ectopic Knox3 gene expression in the primordium of the extra floret is caused by a 305-base pair duplication in intron 4, and phenocopies of the mutation are obtained in the heterologous tobacco system by Knox3 overexpression. It is concluded that homeotic genes of the Knox gene family are involved in floral evocation. Furthermore, the study of polarity of reproductive organs in K and related mutants can now focus on homeobox genes.

[An epidemiological study on unwanted pregnancy]. / Estudio epidemiológico sobre embarazo no deseado.

OBJECTIVE: To define the profile of the woman with an unwanted pregnancy (UWP) and to analyse differences between wanted pregnancies (WP) and UWP. DESIGN: Longitudinal and retrospective observation study. SETTING: La Chana Health Centre, Granada. PATIENTS: 511 pregnancies. MEASUREMENTS AND MAIN RESULTS: Pregnant women were grouped according to whether their pregnancy was WP or UWP. 29.4% of all pregnancies were UWP. 70.5% of pregnancies in under-19s and 75% in over-35s are UWP (chi2 = 12.24; p = 0.00046 regarding the 19 to 35 age group). Women with WP first attend for prenatal care before women with UWP (chi 2 = 10.5; p = 0.0018). CONCLUSIONS: There is a high proportion of UWP, especially among women under 19 and over 35. We defined two profiles of women at risk of UWP.

Molecular cloning and expression patterns of three alleles of the Deficiens-homologous gene St-Deficiens from Solanum tuberosum.

Three alleles of the Deficiens-homologous potato gene St-Deficiens (St-Def) present in the genome of a tetraploid Solanum tuberosum variety were isolated and characterized. For one allele (St-Def pD13) the complete molecular structure was determined by sequence analysis and comparison with its cDNA, while for the other two alleles (pD10, pD12) only partial sequences of regulatory and coding regions were obtained. All three alleles showed (except for one amino acid exchange in pD10) identical sequences in the coding region. While sequence variation was observed within the respective promoters starting some 300 nt upstream of the putative transcriptional start site, the 3' terminal promoter sequences were highly conserved. Within this region, a sequence of 81 nt was identified which showed 73% sequence identity to a corresponding sequence in the Deficiens promoter. This region which contains a putative serum response element was previously shown to regulate the expression of the Deficiens gene in Antirrhinum majus. Expression patterns for the three alleles in transgenic potato lines expressing St-Def promoter/Gus constructs were identical. GUS activity was predominantly located in petals and stamens as expected for the activity of a Def-homologous gene, but a significant level of expression was also detected in the ovary wall and in the vascular bundles supporting anthers and petals. The promoters were also active in abscission structures at the junction of flower stem and pedicel, as well as in anther stomium.

Site-directed mutagenesis and expression in Escherichia coli of WMAI-1, a wheat monomeric inhibitor of insect alpha-amylase.

The wheat monomeric inhibitor WMAI-1 (syn. 0.28) produced in Escherichia coli using the pT7-7 expression vector has the correct N-terminal sequence and the same electrophoretic mobility and specific activity towards the alpha-amylase from the insect Tenebrio molitor as the native WMAI-1 isolated from wheat. This confirms that the native inhibitor is not glycosylated and contradicts claims that a putative glycosyl moiety was essential for inhibition. Thirteen mutants have been obtained at six different sites. Substitution of the highly conserved N-terminal S by the sequence ARIRAR increased the pre-incubation time required for maximum activity. A similar result was obtained by insertion of GPRLPW after position 4, while insertion of EPRAPW at the same position rendered the inhibitor inactive. The substitution D/EGPRL and insertions DGP or D, at position 58, produced complete inactivation. All other mutations had only minor effects on activity.