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Biomedical subjects

H David

Publications and source records attributed to H David.

At least 19 recordsLinked to original sources

Germin-like genes are expressed during somatic embryogenesis and early development of conifers.

Germins and germin-like proteins (GLPs) are members of a superfamily of proteins widely distributed in plants. Their localization within the extracellular matrix and in some cases their hydrogen peroxide-producing activity suggests that these proteins are involved in cell wall metabolism during stress responses and developmental processes. Several very highly conserved conifer GLPs have been identified in somatic embryo tissues. In order to gain more knowledge on their potential involvement in the development of this particular tissue, we have characterized a new GLP gene, LmGER1 in hybrid larch. Anti-GLP immunserum and in-gel activity analyses suggested the presence of superoxide dismutase activity in apoplastic proteins from larch somatic embryos. These results could indicate a possible role for LmGER1 in this physiological process. The expression of LmGER1 has been followed during the maturation of somatic embryos and in different organs of young plantlets by homologous transformation with a promoter-gus construct. This promoter was activated in the root cap of young embryos and, later on, in the cotyledons and in the vascular procambium and xylem. Furthermore, the importance of this gene in embryo development was evaluated by transforming embryonal masses with a gene construct encoding a hairpin RNA leading to gene silencing. The potential role of LmGER1 in cross-linking of cell wall components is discussed.

Amino Acid Sequence↗

ESTs from the fibre-bearing stem tissues of flax (Linum usitatissimum L.): expression analyses of sequences related to cell wall development.

In order to learn more about the diversity of genes expressed during flax fibre cell wall formation, expressed sequence tags (ESTs) were obtained from a cDNA library derived from the outer fibre-bearing tissues of flax (Linum usitatissimum) stems (cv Hermes) harvested at the mid-flowering stage. After elimination of vector and unreadable sequences, 927 ESTs were grouped into 67 clusters and 754 singletons. The flax ESTs have been submitted to the dbEST and GenBank databases with the accession numbers 25939634 - 25940560 (dbEST) and CV478070 - CV478996 (GenBank). Functional analysis allowed the grouping of ESTs into 13 functional categories and revealed that 62 % of ESTs were similar to known sequences, while 12.4 % of ESTs presented no similarity to any known sequences and 25.6 % of ESTs corresponded to proteins of unknown function. The most highly expressed transcripts belonged to four functional categories: protein maturation and metabolism (31 ESTs), signalling (22 ESTs), the cell wall (21 ESTs) and photosynthesis (19 ESTs). 4.4 % (41) of the total ESTs were potentially related to cell wall formation and maturation. The most highly expressed cell wall EST (15 ESTs) corresponded to a beta-xylosidase gene--potentially involved in cell wall remodelling during growth and development. Other cell wall-related ESTs corresponded to cellulose synthase, xyloglucan endotranglucosylase/hydrolase (XTH), beta-galactosidases, and peroxidases. The expression patterns of different cell wall-related ESTs were determined at different developmental stages in flax plants grown under different field conditions. The potential roles of gene products associated with cell wall related ESTs in fibre cell wall development is discussed.

Cell Wall↗

A risk-based methodology for deriving quality standards for organic contaminants in sewage sludge for use in agriculture--Conceptual Framework.

This paper describes a systematic methodology (Conceptual Framework) to derive quality standards for organic (anthropogenic) contaminants in sewage sludge added to agricultural land, in the context of revision of EU Sludge Directive 86/278/EEC and the broader Soil Thematic Strategy. The overall objective is to ensure, based on a risk assessment approach, a sustainable use of sludge over a long time horizon. ILSI-Europe's Conceptual Framework is in essence consistent with the EU Technical Guidance Document (TGD) for Environmental Risk Assessment of Chemicals in the soil compartment, or US-EPA's Sewage Sludge Use and Disposal Regulations, Part 503 Standards. A 'checklist' of different exposure pathways and transfer processes for organic contaminants needs to be considered, and the most sensitive relevant toxicological endpoint and its PNEC need to be identified. The additional complexity specific to deriving Sludge Quality Standards (SQS) is that the toxicity results may need-e.g., for (indirect) human toxicity-to be related back to maximum acceptable soil exposure levels (PEC(soil)). In turn, the latter need to be back-calculated to the maximum acceptable levels in sewage sludge (PEC(sludge)) at the time of application. Finally, for a sustainable sludge use, the exposure from repeated addition and potential chemical build-up over time (e.g., 100 years) needs to be assessed. The SQS may therefore vary with the (local) sludge application regime, and/or sludge pretreatment processes.

Agriculture↗

Simian virus 40 is present in human lymphomas and normal blood.

Many independent studies have demonstrated Simian virus 40 (SV40) in normal and neoplastic human tissues. Clonal integration of virus in the DNA of several thyroid and bone tumors suggests a direct role for SV40 in some cancers. However, in most cases the role of SV40 remains unclear. This study determined the presence of SV40, by amplification followed by hybridization, in 266 normal and neoplastic blood and lymphoid samples. Amplification detected SV40 in 14% of non-autoimmune deficiency syndrome (AIDS) lymphomas, 28% of AIDS related lymphoma and 16% of peripheral blood lymphocytes from non-cancerous patients. No SV40 was detected in leukemia samples. Direct Southern blotting of SV40+ samples detected no virus, consistent with less than one viral genome in ten cells. Sequence analysis of SV40 in blood and lymphoid samples found sequences distinct from laboratory strains of SV40. The presence of limited quantities of SV40 in a small proportion of both normal and neoplastic tissues is suggestive of an adventitious presence with no apparent direct role in blood and lymphoid cancers.

Antigens, Polyomavirus Transforming↗

Rice salT promoter is activated in Papaver somniferum and Nicotiana tabacum transgenic cells in the absence of exogenous ABA.

With the aim of modifying secondary metabolism in Opium poppy (Papaver somniferum) and tobacco (Nicotiana tabacum) cells, gene transfer was performed using the sam1 gene from Arabidopsis thaliana under the control of the salT promoter. This promoter is induced by ABA in rice and in tobacco and we have shown that it is also induced in poppy cells (gus gene). Putatively transformed poppy and tobacco cell lines with the sam1 gene were obtained. In the absence of exogenous inducer we noticed the expression of the transgene resulting in a significant increase of SAM-S activity in all tested transformants of poppy and in half the transgenic tobacco cell lines tested. Addition of ABA to the culture medium failed to enhance the expression of the transgene in both species and resulted in a decrease of the sam1 gene expression in some cell lines. Since the salT promoter is induced by exogenous ABA in both species (gus reporter gene), we suggest a partial sam1 transgene inactivation in certain cell lines. These results show that the efficiency of a regulatory sequence may be different when fused with a reporter gene (gus) compared to fusion with a gene belonging to the housekeeping family (sam1).

Journal Article↗

Rb independent inhibition of cell growth by p15(INK4B).

The INK4 cyclin dependent kinase inhibitors (CDKI), such as p15(INK4B) and p16(INK4A), block cell cycle progression from G to S phase. This is mediated by inhibition of phosphorylation of proteins, including the retinoblastoma susceptibility protein (Rb), by cyclin dependent kinases. Ectopic over-expression of the p16(INK4A) CDKI can inhibit growth of cell lines depending on Rb status. Cell lines lacking Rb, with few exceptions, are resistant to growth inhibition by p16(INK4A). The effects of ectopic over-expression of p15(INK4B) in cell lines with and without wild type Rb were examined by measuring cell recovery. Proliferation was inhibited in cells lacking Rb as well as in cells with wild type Rb expression. Experiments analyzing the effectiveness of chimeric p15(INK4B)/p16(INK4A) proteins indicated that the Rb independent growth inhibition required N-terminal residues of p15(INK4B). Linker insertion mutation of p15(INK4B) showed that the inhibition was dependent on intact ankyrin structures. Double staining flow cytometry found that the growth inhibition correlated with a decrease in cells in G2/M phases of the cell cycle. These findings are consistent with Rb independent inhibition of the progression from G1 to S caused by overexpression of p15(INK4B).

Carrier Proteins↗

Outbreak of multiple drug-resistant tuberculosis in Lisbon: detection by restriction fragment length polymorphism analysis.

SETTING: Multidrug-resistant tuberculosis (MDR-TB) mainly among human immunodeficiency virus (HIV) seropositive patients in Lisbon hospitals in 1996-1997. OBJECTIVE: Detection of transmission of MDR-TB strains and epidemic outbreaks in several hospital units in the city of Lisbon, including a prison hospital. DESIGN: Use of restriction fragment length polymorphism (RFLP) to fingerprint isolates of Mycobacterium tuberculosis resistant to isoniazid, rifampicin, and one other drug. RESULTS: A total of 43 MDR-TB strains were typed. Sixty-seven per cent of the patients were HIV positive, 12% were HIV negative, and the remainder had unknown HIV status. About 88% of the isolates were grouped in three genetically similar clusters, suggesting possible recent transmission. A predominant cluster (cluster A), corresponding to 72% of the cases, was found, 45% of which came from the prison hospital. Strains from this cluster were resistant to isoniazid, rifampicin, streptomycin, and sometimes ethambutol. A retrospective epidemiological investigation was conducted with respect to all patients in cluster A, and epidemiological links were established between them. CONCLUSION: Our results suggest recent transmission of MDR-TB, mainly in HIV-positive patients, in Lisbon hospitals. Moreover, the predominant MDR-TB clustered strains were not confined to HIV-infected individuals, as they were also isolated in some immunocompetent patients.

Cluster Analysis↗

[MRI symptomology in reflex sympathetic dystrophy of the foot].

PURPOSE: To describe the MRI findings of reflex sympathetic dystrophy of the foot and ankle. METHODS: Retrospective study of 50 patients with reflex sympathetic dystrophy of the foot (5 with the cold form, and 45 with the warm form) diagnosed based on clinical and scintigraphic findings. All patients underwent MR imaging. The MRI findings were correlated with the clinical and scintigraphic findings. RESULTS: Patients with the cold form of reflex sympathetic dystrophy had no abnormality of signal at MR imaging. All patients with the warm from of reflex sympathetic dystrophy showed periarticular marrow edema at MR, typically involving more than one bone (mean of 4). Other findings were inconstant: soft tissue edema, joint effusion, and rarely, subchondral band of low T1W signal intensity of unclear etiology. CONCLUSION: MR imaging, including fat-suppressed T2W or STIR images and noncontrast T1W images, is helpful in patients with the warm or acute form of reflex sympathetic dystrophy of the foot. In patients with the cold form, MR imaging is helpful to exclude another underlying etiology for the symptoms and identify patients with the warm form of the process.

Adult↗

Characterization of a germin-like protein gene expressed in somatic and zygotic embryos of pine (Pinus caribaea Morelet).

Germin-like proteins (GLPs) ionically bound to the walls of preglobular somatic embryos of Pinus caribaea Morelet are markers of this early developmental stage. In order to reveal the physiological implications of such markers during early embryo development, we isolated a cDNA clone from somatic embryos predicted to encode a protein with sequence similarity to GLPs. PcGER1 has an open reading frame corresponding to a 220 amino acid polypeptide with a putative N-glycosylation site on Asn-69. The presence of a 24 amino acid putative signal peptide supports the hypothesis of an apoplastic location. The N-terminal 20 amino acid sequence of the predicted mature protein is identical to the amino terminal sequence of GP111, one of the extracellular pine GLPs previously identified. Southern blot hybridizations indicate that PcGER1 is probably unique in the pine genome. Transcripts homologous to PcGER1 are abundant in all embryogenic lines, absent from nonembryogenic lines, and present in quiescent zygotic embryos but not in the female gametophyte, the haploid storage tissue of conifers. Their abundance sharply decreases during germination. Isolation of gf-0.8, a genomic fragment identical to PcGER1 cDNA sequence, confirms that no introns disrupt the coding region as it has been already described for wheat gf-2.8 and gf-3.8 genomic clones. Recombinant PcGER1, produced in Escherichia coli, is recognized by antibodies raised against the GP111 N-terminal nonapeptide and the unglycosylated wheat germin monomer. The implications of GLPs in pine embryogenesis are discussed.

Amino Acid Sequence↗

Preference for sex of first child among women classified as androgynous and nonandrogynous.

This paper examined the relation between gender-role orientation and the preference for sex of firstborn child in 212 pregnant nulliparous women. The Bem Sex-role Inventory was used to assess gender-role orientation of participants. Analysis suggested that gender-role orientation, as measured does not effectively predict the preference for sex of firstborn child.

Adult↗

[MRI in post-traumatic cerebral fat embolism].

We report two cases of posttrauma cerebral fat embolism evaluated with MRI. Due to sensitivity, MRI is the best examination for detecting focal abnormalities of the grey or white matter. MRI helps to make the diagnosis in atypical cases and allows assessment of both extension and course of the embolism.

Accidents, Traffic↗

Evaluation of purine nucleotide loss, lipid peroxidation and ultrastructural alterations in post-hypoxic hepatocytes.

1. Hypoxic alterations in isolated rat hepatocytes were demonstrated by a 90% ATP loss during 60 min of ischaemia and temporary increases of nucleotide degradation products. 2. The oxidative stress during reoxygenation was demonstrated in these cells by a decrease in reduced glutathione (GSH) concentration (30%) and a threefold increase in lipid peroxidation products such as 4-hydroxynonenal and thiobarbituric acid-reactive substances (TBA-RSs). The tremendous GSH loss could not be balanced by the slight oxidized glutathione (GSSG) increase during reoxygenation. 3. For the first time the involvement of free radicals was directly demonstrated using electron spin resonance (ESR) spectroscopy in reoxygenated liver cells. Using the spin trap 5,5-dimethylpyrroline-1-oxide (DMPO), a carbon-centred radical and the adduct of the hydroxyl radical could be detected during early reoxygenation. 4. Morphological alteration of cells was observed, beginning during hypoxia and increasing during post-hypoxic reoxygenation. Electron microscopic findings of hypoxic and post-hypoxic cell damage included pyknosis of nuclei, spherical transformation of mitochondria and increased number of vesicles.

Aldehydes↗

Psychosocial factors related to emotional disturbances during pregnancy.

This study explored a multifactorial model for the understanding of the factors related to the intensity of prenatal emotional disturbances. Data were gathered from 213 pregnant women during the second trimester of pregnancy. Participants were assessed according to a number of psychosocial variables. Two types of prenatal emotional disturbances were examined: depressive symptoms and ambivalence and fears related to maternity. A path analysis indicated that four risk factor domains had a direct effect on depressive symptom level: locus of control; interpersonal relationships; stressors; and psychiatric history. Four risk factor domains also had a direct effect on the level of ambivalence and fears: sociodemographics; depressive symptom level; interpersonal relationships; and family history. Several factors showed an indirect effect on one or both of the disturbances. These results indicate that the study of factors related to prenatal emotional disturbances benefits from a multifactorial model assessing direct and indirect effects.

Adult↗

Xylose-rich polysaccharides from the primary walls of embryogenic cell line of Pinus caribaea.

Embryogenic cell lines of Pinus caribaea were isolated from somatic embryogenesis from zygotic embryos. Previous studies showed that the proteins and glycoproteins were characteristic of the embryogenic state. In the present work we were seeking typical feature in the polysaccharide from the cell walls of embryogenic calli at nine days of culture. Sequential extraction with water, ammonium oxalate, dimethyl sulfoxide, sodium borohydride and 4.3 M potassium hydroxide revealed that the extracted polysaccharides contained high proportions of arabinose and significant amounts of xylose. Fractionation of the hydrosoluble polymers on DEAE cellulose afforded a xylose-rich fraction (80% xylose, 24% glucose and lower properties of fucose and mannose). Methylation analysis and 13C-NMR spectra showed that the glycan backbone consisted of beta 1 --> 4 linked xylosyl residues Similar study of the fractions extracted respectively with DMSO and 4.3 M KOH showed the presence of polydisperse glycoxylans but excluded the presence of xyloglucan in significant amount. This could be a characteristic feature of embryogenic cells walls of Pinus caribaea or could be typical of cells grown as calluses. In the various fractions obtained from DEAE cellulose chromatography of the alkaline extract the infrequent occurrence of fucoxylans beside an arabinogalactan showed again the unusual nature of the cell wall polymers of this embryogenic lines, which seems to differ greatly from those found in the primary wall of cells from suspension cultures.

Carbohydrates↗