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E Hofer

Publications and source records attributed to E Hofer.

At least 37 records · Page 2Linked to original sources
PubMed2000

Model study of vector-loop morphology during electrical mapping of microscopic conduction in cardiac tissue.

The large variety in loop morphology of potential differences recorded at the cardiac surface has been generally attributed to structural discontinuities of the tissue. The aim of this work was to examine if the diversity of vector loops of the electric field E found experimentally may also arise during continuous anisotrope conduction. For this purpose a monodomain computer model was used, consisting of a two-dimensional sheet of excitable tissue surrounded with an unbounded volume conductor. Close to the tissue surface our computations predicted a narrow biphasic course of phi(e) with peak-to-peak separation of less than 400 microm. We examined how accurately E could be reconstructed from measurements recorded with four-element electrode arrays and how activation sequence, interelectrode spacing, and probe orientation affects the results. We found "closed" vector loops of E in planar, and at the apex of elliptical wave fronts, whereas outside of these regions vector loops were "open." Varying probe orientation and size resulted in substantial changes of vector-loop morphology. We concluded that close to the cardiac current sources accurate measurement of E would require interelectrode distances of less than 100 microm.

Animals↗
PubMed2000

Detection of Francisella tularensis in biological specimens using a capture enzyme-linked immunosorbent assay, an immunochromatographic handheld assay, and a PCR.

The early detection of Francisella tularensis, the causative agent of tularemia, is important for adequate treatment by antibiotics and the outcome of the disease. Here we describe a new capture enzyme-linked immunosorbent assay (cELISA) based on monoclonal antibodies specific for lipopolysaccharide (LPS) of Francisella tularensis subsp. holarctica and Francisella tularensis subsp. tularensis. No cross-reactivity with Francisella tularensis subsp. novicida, Francisella philomiragia, and a panel of other possibly related bacteria, including Brucella spp., Yersinia spp., Escherichia coli, and Burkholderia spp., was observed. The detection limit of the assay was 10(3) to 10(4) bacteria/ml. This sensitivity was achieved by solubilization of the LPS prior to the cELISA. In addition, a novel immunochromatographic membrane-based handheld assay (HHA) and a PCR, targeting sequences of the 17-kDa protein (TUL4) gene of F. tularensis, were used in this study. Compared to the cELISA, the sensitivity of the HHA was about 100 times lower and that of the PCR was about 10 times higher. All three techniques were successfully applied to detect F. tularensis in tissue samples of European brown hares (Lepus europaeus). Whereas all infected samples were recognized by the cELISA, those with relatively low bacterial load were partially or not detected by PCR and HHA, probably due to inhibitors or lack of sensitivity. In conclusion, the HHA can be used as a very fast and simple approach to perform field diagnosis to obtain a first hint of an infection with F. tularensis, especially in emergent situations. In any suspect case, the diagnosis should be confirmed by more sensitive techniques, such as the cELISA and PCR.

Antibodies, Monoclonal↗
PubMed1999

Vascular endothelial cell growth factor-induced tissue factor expression in endothelial cells is mediated by EGR-1.

Vascular endothelial cell growth factor (VEGF) is a major regulator of angiogenesis. We report here that treatment of endothelial cells with VEGF leads to upregulation of tissue factor mRNA and protein expression on the cell surface. Reporter gene studies show that transcriptional activation of the tissue factor gene by VEGF is mediated by a GC-rich promoter element containing overlapping binding sites for Sp1 and EGR-1. As shown by immunofluorescence and electrophoretic mobility shift assays, upon VEGF treatment EGR-1 rapidly accumulates in the nucleus and binds to its respective recognition site in the tissue factor promoter. Sp1 occupies this element in unstimulated cells and seems to be partially displaced by increasing amounts of EGR-1. Transfection of endothelial cells with an EGR-1 expression plasmid mimics the upregulation of tissue factor transcription observed after VEGF treatment. In contrast, NFkappaB, the major transcription factor involved in tissue factor upregulation by inflammatory stimuli, is not activated by VEGF. These data show that VEGF induces a response in endothelial cells largely distinct from inflammatory stimuli, and suggest that EGR-1 is a major mediator of the activation of the tissue factor and possibly other VEGF-responsive genes.

Cells, Cultured↗
PubMed1999

Linkage of the NKG2 and CD94 receptor genes to D12S77 in the human natural killer gene complex.

The human natural killer (NK) gene complex is located on the short arm of chromosome 12 and contains a number of genes encoding C-type lectin receptors important for natural killer cell function. Among these are CD94 and the five NKG2 genes. The CD94 protein associates with different NKG2 isoforms to heterodimeric receptors which function to inhibit or trigger cytotoxicity of NK cells depending on the NKG2 isoform. We selected two yeast artificial chromosome clones comprising approximately 1.5 Mb of the NK gene complex and established a contig of underlying P1-derived artificial chromosome clones containing all NKG2 and the CD94 genes. A detailed analysis shows that all six genes are found within a region of 100 to 200 kilobases proximal of the marker D12S77. The gene order established is D12S77 - CD94 - NKG2D - NKG2F - NKG2E - NKG2C - NKG2A. The NKG2 genes are of identical transcriptional orientation, whereas the CD94 gene is placed in opposite orientation. The tight genomic linkage of these genes and the identical orientation of the NKG2 genes suggest coordinate regulation of expression during the differentiation of natural killer cells.

Antigens, CD↗
PubMed1999

Comparison between the role of discontinuities in cardiac conduction and in a one-dimensional hardware model.

In real electrophysiological experiments, irregularities in the extracellular excitation spread are believed to depend on cardiac tissue microstructure. An electronic hardware model was developed to analyze this dependence by placing some inhomogeneities (slow propagation areas) in the medium. The position of such inhomogeneities is correlated with abnormal delays and irregularities measured in signal propagation.

Animals↗
PubMed1999

A transcriptional repressor of the tissue factor gene in endothelial cells.

Tissue factor, the high-affinity receptor and cofactor for the plasma serine protease VII/VIIa, is the primary cellular initiator of the blood coagulation cascade. Inside the vasculature, expression of the tissue factor gene must be tightly controlled. Whereas the endothelium normally does not express tissue factor, on stimulation with inflammatory cytokines or endotoxin the gene is transcriptionally upregulated leading to a procoagulant state. We have now detected a repressive cis-acting element in the tissue factor promoter that downmodulates tissue factor transcription in endothelial cells. In reporter gene assays, deletion of this element leads to an increase of tissue factor transcription and insertion of a trimerized site reduces transcription. Specific protein/DNA complexes are formed on the element with nuclear extracts in electrophoretic mobility shift assays and cross-linking of the proteins followed by SDS-PAGE detects the presence of at least 2 subunits of approximately 40 and 60 kDa, respectively. After transfection of different cell types with the reporter genes, the suppressive effect of the element can only be revealed in endothelial cells. These data suggest that this element represents a novel transcription factor target sequence that functions to suppress expression of the tissue factor gene, preferentially in endothelial cells thereby supporting a noncoagulant state.

Animals↗
PubMed1998

The genomic organization of NKG2C, E, F, and D receptor genes in the human natural killer gene complex.

Interactions of natural killer cell receptors with their cognate ligands play a major role in regulating NK cell function. The NKG2 gene family encodes several highly similar proteins, which are known to form heterodimers with the CD94 receptor. These dimers play a role in the inhibition as well as the activation of NK cells. We have analyzed the gene structures of the NKG2C, D, E, and F genes, and determined their genomic organization. Restriction mapping and sequencing revealed the four genes to be closely linked to one another, and of the same transcriptional orientation. An exon duplication within the NKG2C and E genes was identified, although the duplicated version of this exon has not yet been found in mRNA sequences. The NKG2C, E, and F genes, despite being highly similar, are variable at their 3' ends. We show that NKG2C consists of six exons, whereas NKG2E has seven, and the splice acceptor site for the seventh exon occurs in an Alu repeat. NKG2F consists of only four exons and part of exon IV is in some cases spliced to the 5' end of the NKG2D transcript. NKG2D has only a low similarity to the other NKG2 genes.

Base Sequence↗
PubMed1998

Prevalence of vibrio cholerae O(1) infection in manacapuru, amazonas state, brazil (1992)

This study focused on the prevalence of V. cholerae O(1) infection in 1,196 individuals living in Manacapuru, Amazonas State, through microtitering of vibriocidal antibody and somatic agglutination test. The role of living conditions and individual characteristics as possible risk factors for infection was also assessed. Vibriocidal titers >/= 1: 40 and/or agglutinating titers >/= 1: 80 were considered indicators of V. cholerae O(1) infection. Infection prevalence was 25.7%. There was no significant statistical difference (p=0.05) when analyzed against housing patterns, sanitary facilities, source and treatment of water, destination of domestic waste, sex, or profession. Household location, number of occupants/household, age, and schooling showed significant statistical differences in infection prevalence (p=0.05).

Journal Article↗
PubMed1998

Incidence of Listeria monocytogenes in cheese produced in Rio de Janeiro, Brazil.

The present study evaluated the incidence of Listeria spp. in some Brazilian cheeses obtained from retail stores in Rio de Janeiro, Of 103 samples of various types of cheese examined as recommended in the Listeria isolation protocol of the Health Protection Branch of Canada, 11 (10.68%) were contaminated by Listeria monocytogenes, 13 (12.62%) by Listeria innocua, 6 (5.83%) by Listeria grayi, and 1 (0.97%) by Listeria welshimeri. A higher incidence of L. monocytogenes as observed mainly in the homemade Minas Frescal cheeses (a Brazilian soft white cheese, eaten fresh), 7 of 17 (41.17%), followed by ripened cheeses, 3 of 53 (5.67%), and industrially manufactured Frescal (Minas and Ricotta) cheeses, 1 of 33 (3.03%). Three serotypes (1/2a, 1/2b and 4b) were observed among the strains of L. monocytogenes isolated, all of them being frequently involved in outbreaks of foodborne listeriosis and sporadic cases of the disease all over the world.

Brazil↗
PubMed1997

Adenovirus-mediated expression of green fluorescent protein.

A recombinant replication-deficient adenovirus has been generated that expresses a mutant of the Aquorea victoria green fluorescent protein (GFP) under the control of the strong CMV promoter by insertion into the E1 region (AdV-GFP). High expression of GFP was found in different cell types after infection with the recombinant virus that could be easily detected by fluorescence microscopy. In human umbilical vein endothelial cells (HUVEC), expression levels had already reached a maximum after 2 days and were stable for at least 7 days, as determined by Western blotting. As demonstrated by FACS analysis, up to 98% of HUVEC and approximately 70% of human smooth muscle cells could be transduced to express GFP. Since GFP can be detected in cells without the need for prior fixing and staining, the virus should be useful for optimizing in living cells the transduction efficiency of different cell types, of cells from different experimental animals, as well as studying the kinetics and persistence of adenovirus-mediated gene transfer in diverse experimental settings.

Adenoviridae↗
PubMed1997

Gem, a GTP-binding protein from mitogen-stimulated T cells, is induced in endothelial cells upon activation by inflammatory cytokines.

Using differential screening of cytokine-activated versus resting porcine aortic endothelial cells (PAEC), we have isolated a member of the family of Ras/GTP-binding proteins. The cDNA encodes a 34-kilodalton protein showing 97% homology to Gem, a gene recently isolated from activated T cells, likely representing its porcine homologue. The amino acid sequence differs from the Ras consensus by the absence of a C-terminal isoprenylation site and a glycine to glutamic acid substitution in the third GTP-binding domain. We report here, that pigGem mRNA is strongly inducible in PAEC upon activation by either IL-1 alpha, TNF alpha or lipopolysaccharide (LPS). Low constitutive expression is found in several organs. Epitope-tagged pigGem transfected into endothelial cells (EC) localizes to the cytoplasm and to the inner side of the plasma membrane. Structural features of Gem and its inducibility apparently restricted to T cells and endothelial cells, together with Rad, a GTPase overexpressed in skeletal muscle cells of type II diabetic individuals, define a new branch within the superfamily of GTP-binding proteins.

Amino Acid Sequence↗
PubMed1997

Coliforms and Salmonella in seawater near to domestic sewage sources in Fortaleza (Ceará, Brazil).

The bacteriological conditions of the coastal region of Fortaleza (Ceará, Brasil), including the coastal zones of the Ceará and Cocó rivers, were examined. The study was conducted during June, September and December 1993, and March 1994. The region was divided into two areas: (i) Direct Influence Area (DIA), consisting of 20 sampling stations located near to discharge zones of the submarine pipeline system, where collections were carried out at the surface, and (ii) Indirect Influence Area (IIA), located near to the coastal zone, including Barra do Ceará, Kartódromo, Volta de Jurema, Mucuripe, Farol and Caça e Pesca beaches, totalling 26 sampling stations. The most probable number (MPN) of both total and fecal coliforms in DIA was positive only in station number 6, near to the sewage discharge exit. The following bacteria were identified: Citrobacter sp., Enterobacter aerogenes and Escherichia coli. Kartódromo beach was contaminated throughout the sampling period. Results of total fecal MPN was essentially lower than 3.0 x 10(2) coliforms/100 ml at Caça e Pesca beach. In December, at both DIA and IIA, Salmonella was identified in several samples. In DIA, the spatial distribution for Salmonella suggests that there should be a coastal sea current from east to west along the coastline. In IIA, Salmonella was identified at Kartódromo and Farol beaches throughout the sampling period.

Atlantic Ocean↗
PubMed1996

Influence of the copepod Mesocyclops longisetus (Crustacea: Cyclopidae) on the survival of Vibrio cholerae O1 in fresh water.

In an experimental microcosm, an analysis was performed of the influence exerted by freshwater Mesocyclops longisetus copepods on the survival of Vibrio cholerae O1 serovar Inaba. In the State of Ceará, copepods are used in the control of Aedes aegypti larvae. The system consisted of water with a salinity of 0.27 per 10,000 habitants and pH 7.5, which after sterilizing filtration was distributed into seven flasks with a volume of 400 ml; in each of six flasks, 10 live copepods were inoculated along with 1 ml of an 8-hour culture of Vibrio cholerae O1 at 37oC in Alkaline Peptone Water, resulting in a concentration of 3.80x10(4) colony-forming units. The control flask contained only the water with the same bacterial suspension. The system was maintained for six days at room temperature (25-28oC), and daily duplicate counts were performed in TCBS Agar. Results confirmed a clear association between Vibrio cholerae O1 and the live copepods, based on survival of the bacteria at compatible levels with the initial inoculation until the sixth day of the analysis.

Journal Article↗
PubMed1995

Regulation of the tissue factor promoter in endothelial cells. Binding of NF kappa B-, AP-1-, and Sp1-like transcription factors.

Tissue factor is up-regulated on endothelial cells and monocytes in response to cytokines and endotoxin and is the main trigger of the extrinsic pathway of the coagulation cascade. We have isolated the porcine tissue factor gene and studied the regulation of the promoter, which has not been investigated previously in endothelial cells. Comparison of the promoter sequences with the respective human and murine genes reveals short stretches of homology, which encompass potential binding sites for AP-1, NF kappa B, and Sp1 transcription factors. Using DNase I footprinting, we detect binding of nuclear factors to these promoter elements. Transfection experiments demonstrate that a 300-base pair fragment containing the conserved elements can mediate induced transcription and that the NF kappa B-like element is essential. In accordance, electrophoretic mobility shift assays show a strong increase in the binding of factors to the NF kappa B-like site following induction. We further provide evidence that RelA (p65), c-Rel, and possibly novel polypeptides bind to the tissue factor NF kappa B element. In addition, we show constitutive binding of members of the Fos/Jun and Sp1 families to the AP-1 and Sp1 sites, respectively. We propose a concerted action of AP-1-, NF kappa B-, and Sp1-like factors in transcription from the tissue factor promoter in endothelial cells.

Animals↗
PubMed1995

Intron-exon structure of the porcine I kappa B alpha-encoding gene.

Genomic clones of ECI-6 (endothelial cell inducible), the porcine I kappa B alpha gene that encodes a cytoplasmic inhibitor of the transcription factor NF-kappa B, were isolated, spanning the entire transcribed region plus 2.1 and 0.35 kb of 5'- and 3'-flanking sequences, respectively. The gene contains five introns ranging in size from 0.6 to 0.1 kb. Four of the introns are located in the coding regions for four of the five ankyrin-like repeats in the central part of the I kappa B alpha protein at similar positions. The fifth intron is located in the C-terminal region. Southern blot analysis indicates the presence of a single copy of ECI-6/I kappa B alpha in the porcine genome.

Amino Acid Sequence↗
PubMed1995

NKG2-C is a receptor on human natural killer cells that recognizes structures on K562 target cells.

NKG2-C is a member of the recently discovered NKG2 family of genes and proteins, which are preferentially expressed on human natural killer (NK) cells. These potential NK cell receptors belong to a larger class of type II transmembrane proteins with a C-type lectin domain. We show here that NKG2-C is expressed as a 36-kDa glycoprotein by translation in vitro, recombinant expression and immunoprecipitation from a human NK cell clone. Further, a recombinant soluble NKG2-C-receptor binds specifically to K562 cells, which are target cells for NK cell killing, and to RPMI 8866 cells, which are feeder cells for NK cells; several other hematopoietic cell lines tested do not show any binding. The binding structures on the surface of K562 cells disappear, concomitant with a loss in susceptibility to killing when the cells are induced to differentiate with phorbol ester and Ca2+ ionophore. Our data suggest the presence of specific target molecules for NKG2-C on K562 cells, since overall glycosylation, Lewis X and Lewis Y structures, as well as the mucin-like CD43 molecule, do not change following induction of the cells. We propose that NKG2-C mediates a specific interaction of NK cells and their target cells with functional importance for NK cell killing.

Animals↗
PubMed1994

Haemopoietic cell renewal in radiation fields.

Space flight activities are inevitably associated with a chronic exposure of astronauts to a complex mixture of ionising radiation. Although no acute radiation consequences are to be expected as a rule, the possibility of Solar Particle Events (SPE) associated with relatively high doses of radiation (1 or more Gray) cannot be excluded. It is the responsibility of physicians in charge of the health of astronauts to evaluate before, during and after space flight activities the functional status of haemopoietic cell renewal. Chronic low level exposure of dogs indicate that daily gamma-exposure doses below about 2 cGy are tolerated for several years as far as blood cell concentrations are concerned. However, the stem cell pool may be severely affected. The maintenance of sufficient blood cell counts is possible only through increased cell production to compensate for the radiation inflicted excess cell loss. This behaviour of haemopoietic cell renewal during chronic low level exposure can be simulated by bioengineering models of granulocytopoiesis. It is possible to define a "turbulence region" for cell loss rates, below which an prolonged adaptation to increased radiation fields can be expected to be tolerated. On the basis of these experimental results, it is recommended to develop new biological indicators to monitor haemopoietic cell renewal at the level of the stem cell pool using blood stem cells in addition to the determination of cytokine concentrations in the serum (and other novel approaches). To prepare for unexpected haemopoietic effects during prolonged space missions, research should be increased to modify the radiation sensitivity of haemopoietic stem cells (for instance by the application of certain regulatory molecules). In addition, a "blood stem cell bank" might be established for the autologous storage of stem cells and for use in space activities keeping them in a radiation protected container.

Aerospace Medicine↗