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Biomedical subjects

G Berta

Publications and source records attributed to G Berta.

14 recordsLinked to original sources
PubMed2006

Arbuscular mycorrhizae increase the arsenic translocation factor in the As hyperaccumulating fern Pteris vittata L.

Phytoremediation techniques are receiving more attention as decontaminating strategies. Phytoextraction makes use of plants to transfer contaminants from soil to the aboveground biomass. This research is devoted to study the effects of arbuscular mycorrhizae (AM) on growth and As hyperaccumulation in the Chinese brake fern Pteris vittata. We grew for 45 days P. vittata sporophytes, infected or not infected with the AM fungi Glomus mosseae or Gigaspora margarita, in a hydroponic system on quartz sand. As-treated plants were weekly fed with 25 ppm As. The As treatment produced a dramatic increase of As concentration in pinnae and a much lower increase in roots of both mycorrhizal and control plants. Mycorrhization increased pinnae dry weight (DW) (G. margarita = G. mosseae) and leaf area (G. margarita > G. mosseae), strongly reduced root As concentration (G. mosseae > G. margarita), and increased the As translocation factor (G. mosseae > G. margarita). The concentration of phosphorus in pinnae and roots was enhanced by both fungi (G. margarita > G. mosseae). The quantitatively different effects of the two AM fungi on plant growth as well as on As and P distribution in the fern suggest that the As hyperaccumulation in P. vittata can be optimized by a careful choice of the symbiont.

Arsenic↗
PubMed2002

Down-modulation of nuclear localisation and pro-fibrogenic effect of 4-hydroxy-2,3-nonenal by thiol- and carbonyl-reagents.

Among the oxidative breakdown products of omega-6 unsaturated fatty acids, the aldehyde 4-hydroxy-2,3-nonenal (HNE) is receiving increasing attention for its potential pathophysiological implication, which at least partly lies on the demonstrated ability to modulate gene expression of a number of genes. Here we show that a marked down-modulation of HNE nuclear localisation in cells of a macrophage line (J774-A1) can be afforded by treatment with sulfydryl and carbonyl reagents without significantly interfering with cell viability. As regards the addition of thiol-group reagents to the cell suspension, N-ethylmaleimide (NEM) led to a sustained decrease of HNE nuclear localisation, while 4-(chloromercuri)-benzene-sulfonic acid (PCMBS) gave a similar but more transient effect. Hydroxylamine (HYD), a carbonyl-group reagent, was also able to inhibit HNE nuclear localisation. The actual efficacy of the inhibitors used was then tested on the HNE-induced stimulation of transforming growth factor beta1 (TGFbeta1) production by J774-A1 cells. Indeed, the thiol reagents NEM and PCMBS, both markedly down-modulating HNE nuclear localisation, were able to inhibit HNE-induced increase of TGFbeta1 protein synthesis. The carbonyl reagent HYD was less effective on this respect, producing strong but incomplete protection against HNE-induced TGFbeta1 increase. Taken together, the results indicate that sulfydryl groups are involved in the process of HNE cellular internalisation, while both sulfydryl and carbonyl groups are involved in the process of HNE nuclear translocation, and consequently in the modulation of gene expression by the aldehyde. Further, an actual demonstration is provided that HNE-induced effect on gene regulation can be efficiently counteracted by suitable interference with HNE biochemistry.

Aldehydes↗
PubMed2001

The nucleus of differentiated root plant cells: modifications induced by arbuscular mycorrhizal fungi.

The nuclei of plant cells show marked differences in chromatin organisation, related to their DNA content, which ranges from the type with large strands of condensed chromatin (reticulate or chromonematic nuclei) to one with mostly decondensed chromatin (chromocentric or diffuse nuclei). A loosening of the chromatin structure generally occurs in actively metabolising cells, such as differentiating and secretory cells, in relation to their high transcriptional activity. Endoreduplication may occur, especially in plants with a small genome, which increases the availability of nuclear templates, the synthesis of DNA, and probably regulates gene expression. Here we describe structural and quantitative changes of the chromatin and their relationship with transcription that occur in differentiated cells following an increase of their metabolism. The nuclei of root cortical cells of three plants with different 2C DNA content (Allium porrum, Pisum sativum and Lycopersicon esculentum) and their modifications induced by arbuscular mycorrhization, which strongly increase the metabolic activity of colonised cells, are taken as examples.

Cell Nucleus↗
PubMed2001

Nuclear changes in pathogen-infected tomato roots.

We have investigated nuclear changes induced in tomato (Lycopersicon esculentum) plant roots by two different pathogens, a phytoplasma of the stolbur group and the soil-borne fungus Phytophthora nicotianae var. parasitica, using light and electron microscopy as well as flow cytometry. Our results show that both pathogens strongly influenced nuclear structure, causing pyknosis and chromatolysis, and induced variations in proportion of nuclear populations with different DNA content, increasing the 2C and decreasing the 4C nuclear populations. These latter results suggest a block of the endoreduplication process in the presynthetic phase of the host cell cycle. Moreover, in pathogen-infected roots, nuclei of the same DNA content were less fluorescent, and therefore had a lower DNA content, compared to those of controls, as confirmed by morphological analyses. Our findings suggest that different stimuli may evoke similar nuclear changes, and confirm the usefulness of flow cytometry as a tool to recognize the stress conditions related to the presence of a pathogen.

Animals↗
PubMed2000

[Effectiveness and tolerability of the glucocorticoid mometasone furoate given as nasal spray in seasonal allergic rhinitis].

The clinical effect and tolerance of momethasone furoate (MF) glucocorticoid nasal spray (MFNS) were studied in 14-70 year-old patients suffering from seasonal allergic rhinitis. The patients administered daily one (morning) dose, 100 micrograms each, of MF into both nostrils, for a period of 14 days. They did not use other medicines affecting nasal symptoms. Nasal symptoms (nasal discharge, nasal obstruction, nasal itching, sneezing) and non-nasal symptoms (lacrimation, eye itching/burning sensation, palatal itching, ear itching, general itching), scored 0 to 3, and serving as a basis for evaluating the effect, were registered before treatment (day 1) and at visits on 3, 7 and 14th day. Of the 196 patients involved in the open multicentric study, 188 completed the study. The total average nasal symptom scores decreased, already after 3 days of treatment, from 8.7 to 4.1 and to 1.6 by the 14th day. Decrease of non-nasal symptoms was also conspicuous, however, lacrimation persisted in 57 of 188 cases, while eye itching--mainly in moderate and mild form--in 90 cases. The general condition of rhinitis before the treatment was evaluated by the examiners as severe or very severe in 155 cases (82%), as symptom-free (99 cases) on day 14 in, and mild (71 cases), in 170 cases (90%). The therapeutic effect was considered by both, patients and physician, as excellent, in 106 (56%) and 115 (61%) cases, resp. and as good in 63 (34%) and 56 cases (30%). Side-effects were mostly mild and transitory. Treatment was not discontinued due to side-effect in any of the cases. Based on the results, MFNS, administered in a single daily dose of 200 micrograms, has proved to be an effective and safe glucocorticoid preparation, also easy to use locally, in the treatment of allergic rhinitis.

Administration, Intranasal↗
PubMed1996

Efficiency of plasmid delivery and expression after lipid-mediated gene transfer to human cells in vitro.

Cationic liposome-mediated gene transfer has become increasingly important in the development of experimental therapies for human diseases, such as melanoma, human immunodeficiency virus infection, cystic fibrosis and alpha-1 antitrypsin deficiency. However, very little is known about the mechanisms by which lipid-mediated gene transfer occurs. We studied the kinetics of plasmid delivery and expression by using this technique. Plasmid entry in the cystic fibrosis respiratory epithelial cell line 2CFSME0-1 as well as in two other cell lines (HeP 2g and HeLa) occurred in 95 to 100% of cells within 1 hr of the initiation of lipid-mediated gene transfer. In hepatic and respiratory cells, transcription of a construct containing the cystic fibrosis transmembrane conductance regulator was observed in more than 80% of the cell population; similarly high levels of plasmid utilization were obtained in studies of HLA-B7 expression in human melanoma cells. Studies directly relevant to current human trials of lipid-mediated gene transfer indicate that plasmid entry, transcription and translation are often surprisingly efficient, and may occur in nearly 100% of human cells in culture when sensitive methods for detection are used. Furthermore, conventional X-gal immunohistochemistry markedly underestimates transfection efficiency during transient gene expression. These studies point to a new mechanistic understanding of the features that limit expression by using cationic liposomes.

Cell Line↗
PubMed1995

Arbuscular mycorrhizal induced changes to plant growth and root system morphology in Prunus cerasifera.

We compared root system morphogenesis of micropropogated transplants of Prunus cerasifera L. inoculated with either of the arbuscular mycorrhizal (AM) fungi Glomus mosseae or Glomus intraradices or with the ericoid mycorrhizal species Hymenoscyphus ericae. All plants were grown in sand culture, irrigated with a nutrient solution that included a soluble source of phosphorus, for 75 days after transplanting. Arbuscular mycorrhizal colonization increased both the survival and growth (by over 100%) of transplants compared with either uninoculated controls or transplants inoculated with H. ericae. Arbuscular mycorrhizal colonization increased root, stem and leaf weights, leaf area, root length and specific leaf area, and it decreased root length/leaf area ratio, root/shoot weight ratio and specific root length. Both uptake of phosphorus and its concentration in leaves were increased by AM infection, although the time course of the relationships between intensity of AM infection and P nutrition were complex and suggested a role for factors other than nutrition. The time course for the development of infection varied. It was most rapid with G. mosseae, but it was ultimately higher with G. intraradices. None of the treatments significantly affected the lengths of adventitious roots or the first-, second- or third-order laterals that developed from them. Arbuscular mycorrhizal colonization increased the intensity of branching in all root orders with the effect being most obvious on first-order lateral roots where the number of branches increased from under 100 to over 300 brances m(-1). As a result, although first-order laterals made up 55% of the root systems of control plants, the comparable value was 36% in AM-infected plants. In contrast, second-order laterals represented 25% of control root systems, but 50% of AM-colonized root systems. Glomus intraradices but not G. mosseae increased root diameter. Anatomical studies revealed no changes in the overall form of the root tip, although there were changes in the diameter of the root cap, cell numbers and cell size. Hymenoscyphus ericae increased the duration of the metaphase index. Both AM fungal treatments increased the concentrations of soluble proteins in root extracts and modified the protein profiles by the elimination and addition of protein bands detected by PAGE analysis. We conclude that AM fungal inoculation influenced processes in the root system at different levels, but not all effects were due to improved P nutrition or increased physiological age.

Journal Article↗
PubMed1994

Release of potassium, lipids, and proteins from nonionic detergent treated chicken red blood cells.

The plasma membrane of erythrocytes, as of other cells, is thought to act as the barrier responsible for maintaining intracellular gradients of most ions and small molecular species between the cell and its environment. Controlled application of the nonionic detergent Brij 58 effectively opened the erythrocyte plasma membrane, as judged by electron microscopy and lipid mobilization, but the cytoplasm maintained much of its integrity for about 30 min. Release of K+ correlated well with release of protein into the surrounding medium. The results demonstrate that permeabilization of the erythrocyte plasma membrane does not result in an instantaneous equilibration of small ions, such as K+, between the cell and its environment. A comparison was made between erythrocytes treated with Brij 58 and Triton X-100. The lipid and protein solubilizing actions of Triton X-100 were not as easily separable in time as those of Brij 58. The results of treatment of the erythrocytes with different types of nonionic detergents suggest that the membranolytic and cytoplasmic protein destabilizing actions of nonionic detergents correspond with their hydrophilic-lipophilic balance numbers (HLB values).

Animals↗
PubMed1994

Expression and polarized targeting of a rab3 isoform in epithelial cells.

Pathways of polarized membrane traffic in epithelial tissues serve a variety of functions, including the generation of epithelial polarity and the regulation of vectorial transport. We have identified a candidate regulator of polarized membrane traffic in epithelial cells (i.e., rab3B), which is a member of the rab family of membrane traffic regulators. Rab3B is highly homologous to a brain-specific rab3 isoform (rab3A) that targets in a polarized fashion to the presynaptic nerve terminal, where it probably regulates exocytosis. The coding region for human rab3B was cloned from epithelial mRNA using a reverse-transcription polymerase chain reaction strategy. This cDNA clone hybridized to a single mRNA species in Northern blots of poly(A)+ RNA isolated from epithelial cell lines. A rab3B-specific antibody that was raised against recombinant fusion protein recognized a 25-kD band in immunoblots of cell lysates prepared from cultured epithelial cells (e.g., T84 and HT29-CL19A), but not from a variety of nonepithelial cells (e.g., PC12 neuroendocrine cells). Immunofluorescence analysis confirmed that rab3B protein is preferentially expressed in cultured epithelial cells as well as in a number of native epithelial tissues, including liver, small intestine, colon, and distal nephron. Rab3B localized to the apical pole very near the tight junctions between adjacent epithelial cells within all of these cell lines and native epithelial tissues, as determined by immunofluorescence and immunoelectron microscopic analysis. Moreover, this pattern of intracellular targeting was regulated by cell contact; namely, rab3B was reversibly retrieved from the cell periphery as epithelial cell contact was inhibited by reducing the extracellular Ca2+ concentration. Our results indicate that neurons and epithelial cells express homologous rab3 isoforms that target in a polarized fashion within their respective tissues. The pattern and regulation of rab3B targeting in epithelial cells implicates this monomeric GTPase as a candidate regulator of apical and/or junctional protein traffic in epithelial tissues.

Animals↗
PubMed1993

Rapid purification of recombinant baculovirus using fluorescence-activated cell sorting.

Expression of foreign proteins in the baculovirus-insect cell expression system has been limited by difficulties in rapid identification and purification of recombinant virus. Although the process of identifying recombinant virus has been greatly facilitated by the introduction of vectors that lead to insect cell co-expression of beta-galactosidase with foreign genes of interest, isolation of pure recombinant virus using plaque purification may still take several weeks to months to accomplish. Using a fluorescent beta-galactosidase substrate, we have established that insect cells harboring recombinant virus can be rapidly isolated using fluorescence-activated cell sorting. Pure recombinant virus can then be readily obtained using this cellular fraction, with a pure viral culture generally obtained within 2-3 weeks of insect cell transfection.

Animals↗
PubMed1992

Regulation of plasma membrane recycling by CFTR.

The gene that encodes the cystic fibrosis transmembrane conductance regulator (CFTR) is defective in patients with cystic fibrosis. Although the protein product of the CFTR gene has been proposed to function as a chloride ion channel, certain aspects of its function remain unclear. The role of CFTR in the adenosine 3',5'-monophosphate (cAMP)-dependent regulation of plasma membrane recycling was examined. Adenosine 3',5'-monophosphate is known to regulate endocytosis and exocytosis in chloride-secreting epithelial cells that express CFTR. However, mutant epithelial cells derived from a patient with cystic fibrosis exhibited no cAMP-dependent regulation of endocytosis and exocytosis until they were transfected with complementary DNA encoding wild-type CFTR. Thus, CFTR is critical for cAMP-dependent regulation of membrane recycling in epithelial tissues, and this function of CFTR could explain in part the pleiotropic nature of cystic fibrosis.

Base Sequence↗
PubMed1992

Antisense oligonucleotides to CFTR confer a cystic fibrosis phenotype on B lymphocytes.

Cystic fibrosis transmembrane conductance regulator (CFTR) is expressed at low levels in nonepithelial cells. Recently, we demonstrated that CFTR is responsible for cell cycle-dependent adenosine 3',5'-cyclic monophosphate-responsive Cl- permeability in lymphocytes. Agonist responsiveness of cystic fibrosis (CF) lymphocytes was restored by transfection with plasmid containing wild type CFTR cDNA. CFTR mRNA is expressed in the B lymphoid cell line GM03299; however, quantitative reverse transcriptase-polymerase chain reaction indicates that the level of CFTR mRNA is at least 1,000 times lower than in T84 cells. CFTR protein could not be detected by Western blot or by immunoprecipitation of in vitro phosphorylated protein. However, antisense oligonucleotides representing codons 1-12 of CFTR caused a complete inhibition of cell cycle-dependent Cl-permeability [as determined by 6-methoxy-N-(3-sulfopropyl)-quinolinium fluorescence digital-imaging microscopy], thereby inducing normal cells to acquire a "CF phenotype." These studies provide direct evidence that a CFTR-associated Cl- permeability is present and measurable in lymphocytes, even though CFTR mRNA and protein are expressed at low levels.

B-Lymphocytes↗
PubMedDate not supplied

Ultrastructure of the wall and of the septal pore apparatus in Panaeolus foenisecii (Fr.) Kühn.

The fine structure of the walls and septal pore apparatus in Panaeolus foenisecii is reported. Our description is based on sections of intact hyphae and purified fractions, as well as on the analysis of wall surfaces after purification and shadowcasting, or else of collapsed hypha walls. The cell wall can be more or less homologized to that of the mycelia of Schizophyllum commune; the septal apparatus fits the Polyporus model type.

Agaricales↗