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E Neumann

Publications and source records attributed to E Neumann.

At least 37 records · Page 2Linked to original sources

Cross-language positive priming disappears, negative priming does not: evidence for two sources of selective inhibition.

The authors used a unilingual and bilingual primed lexical decision task to investigate priming effects produced by attended and ignored words. In the unilingual experiment, accelerated lexical decisions to probe target words resulted when the word matched the preceding target word, whereas slowed lexical decisions to probe target words resulted when the word matched the preceding ignored nontarget word. In the bilingual (English-Spanish) experiment, between-language, rather than within-language, priming manipulations were used. Although the ignored repetition negative priming effect replicated across languages, cross-language attended repetition positive priming did not. This dissociation of priming effects in the inter- versus intralanguage priming conditions contradicts episodic retrieval accounts of negative priming that deny the existence of selective inhibitory processes. On the other hand, these results support an extension of inhibition-based accounts of negative priming, because they indicate that inhibition can operate at two levels of abstraction--local word and global language--simultaneously.

Adult↗

Incorporation of the acetylcholine receptor dimer from Torpedo californica in a peptide supported lipid membrane investigated by surface plasmon and fluorescence spectroscopy.

The dimer species (M(r) 580,000) of the nicotinic acetylcholine receptor, isolated from the electric organ of Torpedo californica, was incorporated into a thiopeptide supported lipid bilayer. The incorporation was achieved by fusion of liposomes with reconstituted receptor onto a gold-supported thiopeptide lipid monolayer. Surface plasmon resonance spectroscopy (SPS) was used to monitor in real time the fusion process as well as the specific binding of the antagonist alpha-bungarotoxin. A recently developed extension of SPS offering enhanced sensitivity and specificity, surface plasmon fluorescence spectroscopy (SPFS), was then used to monitor subsequent binding of the monoclonal WF6 and polyclonal antibody, respectively. The latter was fluorescence labeled with Cy5. The different binding assays indicate the successful incorporation of the receptor in the lipid bilayer.

Animals↗

Heterogeneous expression of the tumor-associated antigens RAGE-1, PRAME, and glycoprotein 75 in human renal cell carcinoma: candidates for T-cell-based immunotherapies?

It has recently been shown that tumor-associated antigens (TAAs) can evoke tumor-specific T-cell-defined immune responses in cancer patients, thereby offering the possibility of treating patients with such antigens. To develop T-cell-based immunotherapeutic approaches for renal cell carcinoma (RCC), we studied the mRNA expression profile of the TAAs RAGE-1, tyrosinase, MAGE-1, MAGE-2, NY-ESO-1, Melan-A/MART-1, glycoprotein (gp) 75, gp100, beta-catenin, PRAME, and MUM-1 in 14 human RCC cell lines and in tissue specimens of 37 primary RCCs, 2 related metastases, and 33 specimens of normal renal epithelium. Reverse transcription-PCR was performed with TAA-reactive primers, and the specificity of the PCR products was confirmed by Southern blot and/or direct sequencing. PRAME (10 of 14 cell lines), RAGE-1 (7 of 14 cell lines), and gp75 (4 of 14 cell lines) antigens were expressed in a high percentage of RCC cell lines, although the level of TAA expression varied among the different RCC cell lines. However, low levels of TAA expression in RCC cells are sufficient for recognition by TAA-specific CTLs. Transcription of tyrosinase, Melan-A/MART-1, MAGE-1, MAGE-2, NY-ESO-1, gp100, beta-catenin, and MUM-1 was not detected in any RCC cell line. Approximately 50% of surgically removed neoplasias expressed at least one TAA. RAGE-1 mRNA expression was found in 8 of 39 (21%) RCC samples, PRAME mRNA expression was found in 15 of 39 (40%) RCC samples, and gp75 mRNA expression was found in 4 of 39 (11%) RCC samples, but the expression levels of these TAAs were heterogeneous in the different RCC lesions. One RCC specimen expressed MAGE-2, whereas transcription was not detected in any RCC specimen for MAGE-1, NY-ESO-1, tyrosinase, Melan-A/MART-1, gp100, beta-catenin, and MUM-1. The normal kidney epithelium samples were negative for any TAA tested. Thus, RAGE-1, PRAME, and gp75 expression is found with a different frequency in surgically removed lesions and in RCC cell lines, suggesting that a subgroup of RCC patients could be selected for immunotherapeutic strategies that may benefit from immunization against the RAGE-1, gp75, and/or PRAME antigens. However, additional targets for T-cell-based immunotherapy of RCC have yet to be identified.

Antigens, Neoplasm↗

Discrimination between plasma membrane and intracellular target sites of sphingosylphosphorylcholine.

On the background of the emerging concept of G protein-coupled sphingolipid receptors, Ca2+ mobilization by sphingosylphosphorylcholine (SPPC) in intact cells and SPPC-induced Ca2+ release in permeabilized cells, both occurring at similar, micromolar concentrations, were characterized and compared. In intact human embryonic kidney (HEK-293) cells, SPPC rapidly increased [Ca2+]i by mobilization of Ca2+ from thapsigargin-sensitive stores. In saponin-permeabilized HEK-293 cells, SPPC released stored Ca2+, in a manner similar to but independent of inositol 1,4,5-trisphosphate. Only the action of SPPC on intact cells, but not that in permeabilized cells, was, at least in part, sensitive to pertussis toxin. In addition and most important, Ca2+ release by SPPC in permeabilized cells was not stereoselective, whereas in intact cells only the naturally occurring D-erythro-SPPC, but not L-threo-SPPC, increased [Ca2+]i. Stereoselectivity of SPPC-induced [Ca2+]i increase was also demonstrated in bovine aortic endothelial cells. In conclusion, Ca2+ mobilization by SPPC in intact cells is independent of the previously described SPPC-gated Ca2+ channel on endoplasmic reticulum but probably mediated by a membrane sphingolipid receptor. Thus, SPPC can regulate Ca2+ homeostasis by acting apparently at two cellular targets, which exhibit clearly distinct recognition patterns.

Animals↗

Sphingosine kinase-mediated Ca2+ signalling by G-protein-coupled receptors.

Formation of inositol 1,4,5-trisphosphate (IP3) by phospholipase C (PLC) with subsequent release of Ca2+ from intracellular stores, is one of the major Ca2+ signalling pathways triggered by G-protein-coupled receptors (GPCRs). However, in a large number of cellular systems, Ca2+ mobilization by GPCRs apparently occurs independently of the PLC-IP3 pathway, mediated by an as yet unknown mechanism. The present study investigated whether sphingosine kinase activation, leading to production of sphingosine-1-phosphate (SPP), is involved in GPCR-mediated Ca2+ signalling as proposed for platelet-derived growth factor and FcepsilonRI antigen receptors. Inhibition of sphingosine kinase by DL-threo-dihydrosphingosine and N,N-dimethylsphingosine markedly inhibited [Ca2+]i increases elicited by m2 and m3 muscarinic acetylcholine receptors (mAChRs) expressed in HEK-293 cells without affecting mAChR-induced PLC stimulation. Activation of mAChRs rapidly and transiently stimulated production of SPP in HEK-293 cells. Finally, intracellular injection of SPP induced a rapid and transient Ca2+ mobilization in HEK-293 cells which was not antagonized by heparin. We conclude that mAChRs utilize the sphingosine kinase-SPP pathway in addition to PLC-IP3 to mediate Ca2+ mobilization. As Ca2+ signalling by various, but not all, GPCRs in different cell types was likewise attenuated by the sphingosine kinase inhibitors, we suggest a general role for sphingosine kinase, besides PLC, in mediation of GPCR-induced Ca2+ signalling.

Animals↗

Complication-free duration and the risk of development of retinopathy in elderly diabetic patients.

BACKGROUND: Determining which diabetic patients are at risk for complications and targeting these patients for intensive therapy may avoid the unwanted consequences of hypoglycemia in low-risk patients. Since aging is associated with a decrease in the incidence of diabetic retinopathy, we assessed whether long complication-free duration can define elderly patients at lower risk for future development of diabetic retinopathy. METHODS: In a 10-year clinic-based study, we studied 833 type 2 diabetic patients who were free of diabetic retinopathy and older than 50 years, followed up for more than 4 years. Data included demographic and clinical information on arrival, updated every 3 to 6 months, and yearly direct ophthalmoscopic examination after pupillary dilation by experienced ophthalmologists. All the data were prospectively compiled on relational databases. End points studied were presence of retinopathy, nephropathy, peripheral neuropathy, peripheral vascular disease, hyperlipidemia, and hypertension. RESULTS: Of the patients without retinopathy at the age of 50 years, 10% developed retinopathy during 4 years of follow-up. These patients had longer duration and younger onset of diabetes than the group without retinopathy at the 4-year follow-up. Clustering of microvascular and macrovascular complications was noted. Discriminant analysis showed the following factors to be significant and independent predictors of the development of retinopathy in the elderly: duration of diabetes, body mass index, age, and glucose control. CONCLUSIONS: A long complication-free period does not define elderly patients with type 2 diabetes who are at lower risk for future development of retinopathy. On the contrary, the increase in disease duration is significantly associated with the development of retinopathy in this age group, as described in younger patients.

Aged↗

Mechanism of electroporative dye uptake by mouse B cells.

The color change of electroporated intact immunoglobulin G receptor (Fc gammaR-) mouse B cells (line IIA1.6) after direct electroporative transfer of the dye SERVA blue G (Mr 854) into the cell interior is shown to be dominantly due to diffusion of the dye after the electric field pulse. Hence the dye transport is described by Fick's first law, where, as a novelty, time-integrated flow coefficients are introduced. The chemical-kinetic analysis uses three different pore states (P) in the reaction cascade (C <==> P1 <==> P2 <==> P3), to model the sigmoid kinetics of pore formation as well as the biphasic pore resealing. The rate coefficient for pore formation k(p) is dependent on the external electric field strength E and pulse duration tE. At E = 2.1 kV cm(-1) and tE = 200 micros, k(p) = (2.4 +/- 0.2) x 10(3) s(-1) at T = 293 K; the respective (field-dependent) flow coefficient and permeability coefficient are k(f)0 = (1.0 +/- 0.1) x 10(-2) s(-1) and P0 = 2 cm s(-1), respectively. The maximum value of the fractional surface area of the dye-conductive pores is 0.035 +/- 0.003%, and the maximum pore number is Np = (1.5 +/- 0.1) x 10(5) per average cell. The diffusion coefficient for SERVA blue G, D = 10(-6) cm2 s(-1), is slightly smaller than that of free dye diffusion, indicating transient interaction of the dye with the pore lipids during translocation. The mean radii of the three pore states are r(P1) = 0.7 +/- 0.1 nm, r(P2) = 1.0 +/- 0.1 nm, and r(P3) = 1.2 +/- 0.1 nm, respectively. The resealing rate coefficients are k(-2) = (4.0 +/- 0.5) x 10(-2) s(-1) and k(-3) = (4.5 +/- 0.5) x 10)(-3) s(-1), independent of E. At zero field, the equilibrium constant of the pore states (P) relative to closed membrane states (C) is K(p)0 = [(P)]/[C] = 0.02 +/- 0.002, indicating 2.0 +/- 0.2% water associated with the lipid membrane. Finally, the results of SERVA blue G cell coloring and the new analytical framework may also serve as a guideline for the optimization of the electroporative delivery of drugs that are similar in structure to SERVA blue G, for instance, bleomycin, which has been used successfully in the new discipline of electrochemotherapy.

Animals↗

Membrane electroporation and electromechanical deformation of vesicles and cells.

Analysis of the reduced turbidity (delta T-/T0) and absorbance (delta A-/A0) relaxations of unilamellar lipid vesicles, doped with the diphenylhexatrienyl-phosphatidylcholine (beta-DPH pPC) lipids in high-voltage rectangular electrical field pulses, demonstrates that the major part of the turbidity and absorbance dichroism is caused by vesicle elongation under electric Maxwell stress. The kinetics of this electrochemomechanical shape deformation (time constants 0.1 < or = tau/microsecond < or = 3) is determined both by the entrance of water and ions into the bulk membrane phase to form local electropores, and by the faster processes of membrane stretching and smoothing of thermal undulations. Moreover, the absorbance dichroism indicates local displacements of the chromophore relative to the membrane normal in the field. The slightly slower relaxations of the chemical turbidity (delta T+/T0) and absorbance (delta A+/A0) modes are both associated with the entrance of solvent into the interface membrane/medium, caused by the alignment of the bipolar lipid head groups in one of the leaflets at the pole caps of the vesicle bilayer. In addition, (delta T+/T0) indicates changes in vesicle shape and volume. The results for lipid vesicles provide guidelines for the analysis of electroporative deformations of biological cells.

Cell Membrane↗

Capillaropathy and capillaroneogenesis in the pathogenesis of rosacea.

BACKGROUND: Dilatation of vascular vessels in rosacea has generally been attributed to yielding to deranged connective tissue. In contrast, in a previous study a degrading effect of insufficient vascular vessels in connective tissue has been demonstrated in connection with diabetic microangiopathy. In yet another paper, it was demonstrated that the fusing of damaged capillaries, which had lost part of their adjacent walls due to functional inadequacy, led to the formation of dilated vessels. These pathogenetic patterns are the main subjects of this study of rosacea. METHODS: Punch biopsies from rosacea lesions were examined by light microscopy and immunochemistry in order to identify structural changes leading to the formation of telangiectasias. RESULTS: Deranged connective tissue is secondary to damaged capillaries. The primary damage may be evoked mostly by environmental influences, mainly the sun. CONCLUSIONS: Infections, infestations, and granulomatous formations are not primary in the development of rosacea.

Adult↗

Parasitic infections in germfree animals.

The association of vertebrate hosts with the indigenous microbiota and its effect on the response to infections has long been a subject of scientific curiosity. From the first theory supported by Louis Pasteur that life would be impossible in the absence of associated microorganisms to the development of germfree mammals for research, a lot was learned about how the normal microbiota influences the environment in which pathogens may find themselves. In the present review, we attempt to summarize the more recent results from our group and others on the influence of the normal microbiota on the outcome of parasitic infections. Our results and those of others point to a complex relationship between the mammalian system and its indigenous microbiota, leading to greater resistance to some infections and enhanced susceptibility to others.

Animals↗

Monoassociation with Lactobacillus acidophilus UFV-H2b20 stimulates the immune defense mechanisms of germfree mice.

Probiotics are formulations containing live microorganisms or microbial stimulants that have some beneficial influence on the maintenance of a balanced intestinal microbiota and on the resistance to infections. The search for probiotics to be used in prevention or treatment of enteric infections, as an alternative to antibiotic therapy, has gained significant impulse in the last few years. Several studies have demonstrated the beneficial effects of lactic acid bacteria in controlling infection by intestinal pathogens and in boosting the host's nonspecific immune response. Here, we studied the use of Lactobacillus acidophilus UFV-H2b20, a lactic acid bacterium isolated from a human newborn from Viçosa, Minas Gerais, Brazil, as a probiotic. A suspension containing 10(8) cells of Lactobacillus acidophilus UFV-H2b20 was inoculated into groups of at least five conventional and germfree Swiss mice to determine its capacity to stimulate the host mononuclear phagocytic activity. We demonstrate that this strain can survive the stressing conditions of the intestinal tract in vivo. Moreover, the monoassociation of germfree mice with this strain for seven days improved the host's macrophage phagocytic capacity, as demonstrated by the clearance of a Gram-negative bacterium inoculated intravenously. Monoassociated mice showed an undetectable number of circulating E. coli, while 0.1% of the original inoculum was still present in germfree animals. Mice treated with viable or heat-killed Lactobacillus acidophilus UFV-H2b20 presented similarly improved clearance capacity when compared with germfree controls. In addition, monoassociated mice had twice the amount of Kupffer cells, which are responsible for the clearance of circulating bacteria, compared to germfree controls. These results suggest that the L. acidophilus strain used here stimulates a nonspecific immune response and is a strong candidate to be used as a probiotic.

Animals↗

[Fluticasone propionate, corticosteroid for topical treatment].

The aim of this study was to evaluate the usefulness of fluticasone propionate in various dermatoses and to present the data to non-dermatologists. The study was performed at the Department of Dermatology, University School of Medicine in Poznań. The studied group consisted of 40 cases of various types of eczema, psoriasis vulgaris, DLE, photodermatoses and morphea. Fluticasone propionate cream or ointment was applied to lesional skin twice daily for 7-19 days. Acute eczematous skin lesions significantly improved after the first seven days of treatment, whereas in patients with chronic eczema the good therapeutic results were observed after 14-19 days. Fluticasone propionate was also effective in the treatment of DLE and photodermatoses.

Administration, Topical↗

Sphingosine-mediated electroporative DNA transfer through lipid bilayers.

When the cationic sphingosine is present in planar lipid bilayers, the adsorption of pDNA is enhanced and the electroporatability of the bilayer is facilitated. Furthermore, pDNA, adsorbed to lipid bilayers composed of diphytanoyl lecithin and D-sphingosine causes electroporative channel-like events of conductance 1 to 7 pS, provided the voltage polarity is correct and the voltage is high enough, Vm > or = +30 mV. The data suggest electrotransport of pDNA through the bilayer, mediated by transient complexes between DNA and the lipids in the pore edges of elongated, electropercolated hydrophilic pore zones. The results are the basis for the optimisation of electroporative transfer of DNA or oligonucleotides to cells and tissue.

DNA↗

Annexin V and vesicle membrane electroporation.

The method of membrane electroporation (ME) has been used as an analytical tool to quantify the effect of membrane curvature on transient electric pore formation, and on the adsorption of the protein annexin V (M(r)= 35,800) to the outer surface of unilamellar lipid vesicles (of radii 25 < or = a/nm < or = 200). Relaxation kinetic studies using optical membrane probes of the diphenylhexatriene type suggest that electric pore formation is induced by ionic interfacial polarization causing entrance of the (more polarizable) water into the lipid bilayer membrane yielding (hydrophobic and hydrophilic) pore states with a mean stationary pore radius rp = 0.35 (+/-0.05) nm. Extent and rate of ME, compared at the same induced transmembrane voltage, were found to decrease both with increasing vesicle radius and with increasing protein concentration. This 'inhibitory' effect of annexin V is apparently allosteric and saturates at about [ANT]sat = 4 microM annexin V for vesicles of a = 100 nm at 1 mM total lipid concentration, 0.13 mM total Ca2+ concentration and at T = 293 K. Data analysis in terms of Gibbs area-difference-elasticity energy suggests that the bound annexin V reduces the gradient of the lateral pressure across the membrane. At [ANT]sat, about 20% of the vesicle surface is covered by the bound protein, but it is only 0.01% of the surface of the outer lipid leaflet in which a part of the protein, perhaps the aromatic residue of the tryptophan (W 187), is inserted. Insertion leads to a denser packing of the lipid molecules in the outer membrane leaflet. As a consequence, the radius of the electropores in the remaining membrane part, not covered by annexin V decreases (rp/nm = 0.37, 0.36 and 0.27) with increasing adsorption of the protein ([ANT] = 0, 2 and 4 microM, respectively).

Adsorption↗

A time course analysis of Stroop interference and facilitation: comparing normal individuals and individuals with schizophrenia.

Using randomized stimulus onset asynchrony (SOAs), the authors traced the time course of Stroop interference and facilitation in normal participants and participants with schizophrenia. Unlike earlier findings using blocked SOAs, singular peaks in interference, facilitation, or both occurred at particular SOAs. The peaks of normal participants and participants with schizophrenia differed. Findings are congruent with a model of Stroop performance that posits individual differences in processing speeds of target and nontarget stimulus dimensions, coupled with critical points in response selection. Participants with schizophrenia also showed more overall interference than normal control participants. A second experiment that added a temporal gap between the distractor word and target color obliterated Stroop effects only for individuals with schizophrenia. These findings provide a new empirical basis for models of Stroop effects. They are also consistent with hypotheses about the importance of the prefrontal cortex for working memory and prefrontal dysfunction in schizophrenia.

Adolescent↗

Stroop theory, memory, and prefrontal cortical functioning: reply to Cohen et al. (1997)

In this article, the authors respond to J. D. Cohen, K. O. Dunbar, D. M. Barch and T. S. Braver's (1997) comment on their target article. The present article (a) takes issue with the characterization given by Cohen et al. of the authors' approach as a classical speed-of-processing account of Stroop effects, (b) discusses the value and relevance of other theoretical concepts, such as traditional accounts of working memory and parallel distributed processing (PDP) approaches to Stroop effects, (c) further examines the possibility that the differences the authors observed in Experiment 2 between normal participants' performance and that of participants with schizophrenia may have been due to distractor stimulus degradation, particularly for participants with schizophrenia, and (d) argues for the relevance of the prefrontal cortex, as well as other brain areas, in accounting for their results. The authors conclude with a final theoretical question.

Attention↗