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W Richter

Publications and source records attributed to W Richter.

At least 37 records · Page 2Linked to original sources
PubMed2001

Functional magnetic resonance imaging of visual object construction and shape discrimination : relations among task, hemispheric lateralization, and gender.

We studied the brain activation patterns in two visual image processing tasks requiring judgements on object construction (FIT task) or object sameness (SAME task). Eight right-handed healthy human subjects (four women and four men) performed the two tasks in a randomized block design while 5-mm, multislice functional images of the whole brain were acquired using a 4-tesla system using blood oxygenation dependent (BOLD) activation. Pairs of objects were picked randomly from a set of 25 oriented fragments of a square and presented to the subjects approximately every 5 sec. In the FIT task, subjects had to indicate, by pushing one of two buttons, whether the two fragments could match to form a perfect square, whereas in the SAME task they had to decide whether they were the same or not. In a control task, preceding and following each of the two tasks above, a single square was presented at the same rate and subjects pushed any of the two keys at random. Functional activation maps were constructed based on a combination of conservative criteria. The areas with activated pixels were identified using Talairach coordinates and anatomical landmarks, and the number of activated pixels was determined for each area. Altogether, 379 pixels were activated. The counts of activated pixels did not differ significantly between the two tasks or between the two genders. However, there were significantly more activated pixels in the left (n = 218) than the right side of the brain (n = 161). Of the 379 activated pixels, 371 were located in the cerebral cortex. The Talairach coordinates of these pixels were analyzed with respect to their overall distribution in the two tasks. These distributions differed significantly between the two tasks. With respect to individual dimensions, the two tasks differed significantly in the anterior--posterior and superior--inferior distributions but not in the left--right (including mediolateral, within the left or right side) distribution. Specifically, the FIT distribution was, overall, more anterior and inferior than that of the SAME task. A detailed analysis of the counts and spatial distributions of activated pixels was carried out for 15 brain areas (all in the cerebral cortex) in which a consistent activation (in > or = 3 subjects) was observed (n = 323 activated pixels). We found the following. Except for the inferior temporal gyrus, which was activated exclusively in the FIT task, all other areas showed activation in both tasks but to different extents. Based on the extent of activation, areas fell within two distinct groups (FIT or SAME) depending on which pixel count (i.e., FIT or SAME) was greater. The FIT group consisted of the following areas, in decreasing FIT/SAME order (brackets indicate ties): GTi, GTs, GC, GFi, GFd, [GTm, GF], GO. The SAME group consisted of the following areas, in decreasing SAME/FIT order : GOi, LPs, Sca, GPrC, GPoC, [GFs, GFm]. These results indicate that there are distributed, graded, and partially overlapping patterns of activation during performance of the two tasks. We attribute these overlapping patterns of activation to the engagement of partially shared processes. Activated pixels clustered to three types of clusters : FIT-only (111 pixels), SAME-only (97 pixels), and FIT + SAME (115 pixels). Pixels contained in FIT-only and SAME-only clusters were distributed approximately equally between the left and right hemispheres, whereas pixels in the SAME + FIT clusters were located mostly in the left hemisphere. With respect to gender, the left-right distribution of activated pixels was very similar in women and men for the SAME-only and FIT + SAME clusters but differed for the FIT-only case in which there was a prominent left side preponderance for women, in contrast to a right side preponderance for men. We conclude that (a) cortical mechanisms common for processing visual object construction and discrimination involve mostly the left hemisphere, (b) cortical mechanisms specific for these tasks engage both hemispheres, and (c) in object construction only, men engage predominantly the right hemisphere whereas women show a left-hemisphere preponderance.

Adult↗
PubMed2001

Ranking fMRI time courses by minimum spanning trees: assessing coactivation in fMRI.

In fMRI, time courses with similar temporal "activation" patterns may belong to different brain regions (i.e., these regions are functionally connected, coactivated). A group of time courses (TCs) corresponding to a particular type of temporal activation pattern should be maximally self-consistent (homogeneous). We demonstrate that ordering a group of multidimensional fMRI time courses by a minimum spanning tree (MST) may be used to investigate the temporal homogeneity of a group of TCs. We show the utility of MST ranking for data-driven analysis methods in investigating coactivation in fMRI. MST ranking is equally useful for hypothesis-led methods. Furthermore, MST ranking enables pairwise comparisons of groups/clusters (i.e., any collection of TCs, no matter how derived) of fMRI time courses.

Brain↗
PubMed2001

Distraction modulates anterior cingulate gyrus activations during the cold pressor test.

The anterior cingulate gyrus (ACG) is part of a neural network implicated in attention-demanding tasks, such as the experience of pain. However, the regions within the ACG responding to cognitive demands and to painful stimulation are not identical. Since directing attention away from a painful stimulus is known to reduce the perceived pain intensity, we hypothesized that distraction from pain would result both in decreased activation of ACG subregions responsive to painful stimulation and increased activation of ACG subregions responsive to the distraction task. BOLD fMRI has comparatively high spatial resolution and allows for better identification of ACG subregional responses than other neuroimaging techniques. Twelve subjects were tested using the cold pressor test (CPT), a verbal attention task (VAT), and a distraction task (DT) (a combination of the CPT and VAT). Analysis was performed on a voxel-by-voxel basis using a general linear model as implemented in SPM99. In addition to ACG activations common to both the CPT and VAT, we identified one CPT-specific cluster in an area corresponding to BA24'. The modulation effect of distraction on pain was assessed by contrasting (CPT-DT) and (DT-CPT). In support of our hypothesis, contrast (CPT-DT) revealed a decrease in BA24' during the DT and contrast (DT-CPT) showed increased activation in BA32/32'. These data suggest that distraction from pain and concomitant low pain ratings are reflected in distinct ACG subregional responses.

Adult↗
PubMed2001

[A structured pain interview for geriatric patients].

BACKGROUND: In old age, assessment of pain often is hampered by sensory and cognitive deficits that do not allow the patients to fill in standardized questionnaires without help from significant others. Therefore, as an alternative, we developed a structured pain interview, and examined its properties and acceptance in a sample of geriatric patients with pain. METHODS: The interview covers site of pain, intensity of pain, its duration and persistency, pain related disability and, finally, emotional and cognitive variables. In addition, the interviewer addresses significant others to get information about medication, previous treatment, and residence, and administers the Mini-Mental-State-Examination (MMSE). The analysis includes 128 patients of pain centers older than 74 years, of whom 80% are female. RESULTS: Forty percent of the subjects score below the critical MMSE value < or =23 indicative of cognitive impairment. These patients are responsible for 36 out of a total of 39 missing values. A significant increase of missing values is observed in patients with a MMSE score below 10. Cognitive impairment goes along with greater functional and social disability. On the other hand, cognitive impairment is unrelated to localization, intensity, and duration of pain. The memory item of the MMSE can be used as a screening tool for cognitive impairment. Patients, who are unable to recall any of the three objects, comprise 80% of the total of missing values and demonstrate a low MMSE score. CONCLUSION: As long as geriatric patients are able to communicate verbally, they are most likely to profit from the structured pain interview in spite of existing cognitive impairment. A MMSE score <10 indicates that the interpretation of the data obtained may be difficult, especially due to a high frequency of missing values.

Aged↗
PubMed2001

Identification of substrate specificity determinants in human cAMP-specific phosphodiesterase 4A by single-point mutagenesis.

To identify amino acids that might be involved in discriminating guanosine-3',5'-cyclic phosphate (cGMP) towards adenosine-3',5'-cyclic phosphate (cAMP) binding in the cAMP-specific phosphodiesterases, alignments of different human cyclic nucleotide phosphodiesterases (PDEs) were performed. Eight amino acid residues that are highly conserved in the cAMP-hydrolysing phosphodiesterases (PDE1, PDE3, PDE4, PDE7, PDE8) and that did not show any homologies to the cGMP-specific phosphodiesterases (PDE5, PDE6, PDE9) were selected from these alignments. Using the technique of site-directed mutagenesis, derivatives of PDE4A carrying single mutations at these conserved residues (amino acid positions are given according to the human PDE4A isoform HSPDE4A4B; accession number L20965) were generated and expressed in COS1 cells. The expression products were characterised with regard to cAMP and cGMP hydrolysis and sensitivity towards type-specific inhibitors. The mutation of Phe484 toward Tyr, Ala590 toward Cys, Leu391 and Val501 towards Ala had no significant influence on substrate affinity or specificity. However, the exchange of Trp375 and Trp605 for aliphatic residues abolished catalytic activity and the exchange of Pro595 for Ile led to sevenfold decrease of substrate affinity and an 14-fold decrease of the affinity towards the PDE4-specific inhibitor 4-[3-(cyclopentoxyl)-4-methoxyphenyl]-2-pyrrolidone (rolipram). Both effects may provide evidence for a structural importance of Trp375, Trp605 and Pro595 for PDE function. By exchanging the aspartate residue for asparagine or alanine at position 440 of the human PDE4A4B isoform, the substrate specificity was altered from the highly specific cAMP hydrolysis to an equally efficient cAMP and cGMP binding and hydrolysis. In addition, the IC(50) values for common PDE4-specific inhibitors like rolipram, N-(3,5-dichlorpyrid-4-yl)-3-cyclopentyl-oxy-4-methoxy-benzamide (RPR-73401) and 8-methoxy-5-N-propyl-3-methyl-1-ethyl-imidazo[1,5-a]-pyrido[3,2-e]-pyrazinone (D-22888) were dramatically increased. These results demonstrate an important role of the aspartate at position 440 in determining substrate specificity and inhibitor susceptibility of PDE4A. The strong conservation of this residue suggests that Asp440 may play a similar role in other cAMP-PDEs.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed2001

Identification of inhibitor binding sites of the cAMP-specific phosphodiesterase 4.

Using the technique of site-directed mutagenesis, point mutants of human PDE4A have been developed in order to identify amino acids involved in inhibitor binding. Relevant amino acids were selected according to a peptidic binding site model for PDE4 inhibitors, which suggests interaction with two tryptophan residues, one histidine and one tyrosine residue, as well as one Zn(2+) ion. Mutations were directed at those tryptophan, histidine, and tyrosine residues, which are conserved among the PDE4 subtypes (PDE4A-D) and lie within the high-affinity 4-[3-(cyclopentoxyl)-4-methoxyphenyl]-2-pyrrolidone (rolipram) binding domain of human PDE4A (amino acids 276-681 according to the PDE4A sequence L20965). Truncations to this region do not alter enzyme activity or inhibitor sensitivity. The mutants were expressed in COS1 cells, and the recombinant cyclic nucleotide phosphodiesterase (PDE) forms have been characterized in terms of their catalytic activity and inhibitor sensitivities. Tyrosine residues 432 and 602, as well as histidine 588, were found to be involved in inhibitor binding, but no interaction was detected between tryptophan and PDE inhibitors tested. To test the possibility that other amino acids are of importance for hydrophobic interactions, selected phenylalanine residues were also mutated. We found phenylalanine 613 and 645 to influence inhibitor binding to PDE4. The significant differences in the inhibitor sensitivities of the mutants show that the various inhibitors have different enzyme binding sites. Based on the assumption that the known side effects of PDE4 inhibitors (like emesis and nausea) are caused directly by selective inhibition of different conformation states of PDE4, our results may be a hint to differ between PDE4 inhibitors, which have emetic side effects (like rolipram), and those that do not have side effects (like N-(3,5-dichlorpyrid-4-yl)-[1-(4-fluorbenzyl)-5-hydroxy-indol-3-yl]-glyoxylateamide [AWD12-281]) by the differences of their binding sites and in that context contribute to the development of novel drugs. Furthermore, the identification of amino acid interactions proposed by the peptidic binding site model, which was used for the mutant selection, verifies the PrGen modeling as a useful method for the prediction of inhibitor binding sites in cases where detailed knowledge of the protein structure is not available.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed2001

Freeze-fracture studies on lipids and membranes.

Freeze-fracture electron microscopy is especially useful for investigation of lipid structures by the advantageous fracture course within hydrophobic zones. Freezing is, on the other hand, a restriction because the structures of lamellar and non-lamellar phase states with disordered acyl chains (L(alpha), H(II,) cubic) are difficult to preserve. An important aspect of this method is therefore the lipid structure of phase states with ordered acyl chains (crystal, gel), and with a different degree of hydration. Freeze-fracture of pure lipid systems creates a valid representation of the structure of non-lamellar phases and of the general structure of the "lamellar" lipid bilayer, and lamellar phases with characteristic deformations (ripples, curvatures, plane sectors) can be identified. Fracture through the hydrophobic bilayer centre of biological membranes reveals characteristic protein components, the intramembraneous particles (IMPs). The lateral distribution of the IMPs is a helpful marker for fluid and rigid phase states, also without deformation of the lamella. The overall history and the present state of knowledge concerning the different structures revealed by the freeze-fracture and freeze-etch techniques in lipid systems, and to a limited extent in biological membranes, is reviewed, taking into account studies from our own laboratory.

Animals↗
PubMed2001

Uptake and transfer of PCDD/Fs by cattle fed naturally contaminated feedstuffs and feed contaminated as a result of sewage sludge application. 2. Nonlactating cows.

The dietary absorption and tissue distribution of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) was investigated in 4 nonlactating Simmental cows. During Phase 1 the dietary uptake and fecal excretion of these chemicals were measured over 10 days using feed containing background levels of PCDD/Fs that were primarily of atmospheric origin. Following this, two of the animals were sacrificed and samples of different fat, muscle, and organ tissues were collected. In Phase 2 the remaining two animals were fed grass silage from a field which had a history of repeated sewage sludge applications. During the last 10 days of the 27-day feeding period, the dietary uptake and fecal excretion of PCDD/Fs were again quantified, after which these two animals were also sacrificed and sampled. The dietary absorption of the PCDD/Fs in the nonlactating cows agreed well with values reported in Part I of this series for lactating cows. In the two animals sacrificed at the end of Phase 1 that were close to a contaminant steady state, the lipid-normalized concentrations were similar in almost all tissues. The exceptions were the liver, and to a lesser extent the lungs and the spleen, which had higher levels; and the degree of elevation increased with the degree of chlorination of the PCDD/Fs. During Phase 2, the animals' body burden of several of the PCDD/F congeners increased markedly. The tissue analyses indicated that the chemicals were initially sequestered primarily in the liver, from where they were redistributed to the other tissues and organs. The rate of redistribution was related to the perfusion of the organ/tissue and decreased in the order lung>spleen>kidney>muscle>fat tissue. The rate of redistribution also decreased with increasing degree of chlorination of the PCDD/F congeners. Whereas virtually all of the 1,2,3,7,8-Cl(5)DD taken up during Phase 2 had been deposited in fat tissue by the end of the 27-day feeding period, three-quarters of the Cl(8)DD was still in the liver.

Animal Feed↗
PubMed2001

The structure of Si nanocrystals on SiC.

Si nanocrystals grown on cubic SiC have been characterized using high-resolution transmission electron microscopy. At lower temperatures nanocrystals grow in two different orientations, whereas at higher temperatures they grow in a single preferred orientation. The nanocrystals are shown to be unstrained; in some cases possibly due to the presence of a thin amorphous wetting layer.

Letter↗
PubMed2001

Isolation and functional characterization of recombinant GAD65 autoantibodies derived by IgG repertoire cloning from patients with type 1 diabetes.

The generation of human monoclonal autoantibodies is critical for understanding humoral immune responses in autoimmunity. In this study, we isolated the first human recombinant antibodies to glutamate decarboxylase (rGAD65ab) by IgG repertoire cloning, phage display of Fab fragments, and biopanning from two patients at onset of type 1 diabetes. We demonstrate that natural Ig heavy- and light-chain pairings of autoantibodies can be isolated by the recombinant approach and have a major selection advantage over other rGAD65ab. Among eight rGAD65ab, three (rGAD65ab A-C) displayed all functional and structural properties of known disease-related GAD65ab, including reactivity in the enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), islet cell antibody (ICA) test, and variable gene usage. Dominant epitope recognition was directed to the previously defined epitope EP-1 in the middle of GAD65, corroborating its immunodominance in the molecule. New features, such as assay-dependent GAD65 reactivity and new epitope recognition, were observed in two rGAD65ab (D and E). These antibodies were positive in the GAD65 ELISA and ICA test but not in the GAD65 RIA, providing the first examples for ICA with incongruent results in solid-phase and fluid-phase assays. In conclusion, phage display-derived antibodies reflected well the natural autoantibody response in type 1 diabetes and may allow further characterization of assay-dependent features of GAD65ab and the recognition of epitopes in solid- but not fluid-phase assays.

Adult↗
PubMed2001

A growth and differentiation factor-5 (GDF-5)-coated suture stimulates tendon healing in an Achilles tendon model in rats.

Growth and differentiation factor-5 (GDF-5) is essential for normal skeletal development and induces tendon-and ligament-like structures at ectopic sites. Therefore, we investigated the influence of a GDF-5-coated suture on the healing Achilles tendon in rats. The right Achilles tendon in 80 rats was transected and sutured with an absorbable polyglactin suture. Animals were randomized to an uncoated-suture control and a GDF-5-coated suture group. At 1, 2, 4 and 8 weeks after surgery the repair tissue was evaluated biomechanically and histologically. Biomechanical testing revealed significantly thicker tendons, which were stiffer at 1, 2, and 4 weeks, in the experimental group than in the control group. The maximum tensile strength was significantly increased at 2 weeks after surgery. Histologically we found cartilage-like cell nests 4 weeks after tendon repair, which were positive for type II collagen. In conclusion, local growth factor delivery by a coated suture material showed a promising beneficial effect on tendon repair. The appearance of cartilage-like structures may demonstrate the chondroinductive capacity of GDF-5, which in these circumstances, however, might be overcome by modifications of the GDF-5 dose and/or the suture material.

Achilles Tendon↗
PubMed2001

Bone morphogenetic protein-2 and growth and differentiation factor-5 enhance the healing of necrotic bone in a sheep model.

INTRODUCTION: Osteotropic growth factors enhance bone repair, but their efficacy in an area of necrotic bone is not known. The purpose of this study was to investigate the effects and potential side effects of an intraosseous application of absorbable bone morphogenetic protein-2 (BMP-2) and growth and differentiation factor-5 (GDF-5) composites in a sheep model for partial necrosis of the femoral head. MATERIALS AND METHODS: The direct injection of ethanol under fluoroscopy into the superior centre of the right femoral head produced histologically documentable necrosis of the central region of the head in a previous study of ten sheep. Another 27 sheep constituted the sample to study the effects of BMP-2 and GDF-5. Necrosis was produced in the same fashion in these animals. Four weeks later nine sheep received 300 microg recombinant BMP-2 and nine sheep 300 microg recombinant GDF-5 on an absorbable carrier by surgical implantation. Nine sheep received the carrier alone (control group). The animals were sacrificed at 3, 6, and 12 weeks after implantation and both femora were harvested. RESULTS: Bone density analysis and microscopic examination indicated that bone formation was noticeably induced as early as 3 weeks postoperatively in the growth factor treated animals. Bone regeneration was enhanced by growth factor composites. This was documented by histological scoring and histomorphometric analysis. No severe local side effects secondary to the growth factors, such as heterotopic ossification or inflammation, were observed in either group. DISCUSSION: The application of an absorbable growth factor composite in combination with established surgical techniques is a promising approach, that may enhance the healing of devitalised bone defects. Based on these results, further studies regarding biodegradation, dosage of the protein and surgical technique are required.

Animals↗
PubMed2001

Fusion proteins for combined analysis of autoantibodies to the 65-kDa isoform of glutamic acid decarboxylase and islet antigen-2 in insulin-dependent diabetes mellitus.

BACKGROUND: Prediction, risk assessment, and diagnosis of autoimmune diseases often rely on detection of autoantibodies directed to multiple target antigens, such as the 65-kDa isoform of glutamic acid decarboxylase (GAD65-abs) and the tyrosine phosphatase-like protein islet antigen-2 (IA2-abs), the two major subspecificities of islet cell antibodies (ICAs) associated with insulin-dependent diabetes mellitus. We hypothesized that a combination of autoantigens in a fusion protein unifying the important immunodominant epitopes could provide an efficient target for cost-effective, one-step screening of sera. METHODS: Chimeric proteins composed of GAD65 and IA2 residues were constructed, analyzed for their immune reactivity with monoclonal antibodies and sera, and used in a diagnostic assay with (35)S-labeled protein as antigen. RESULTS: Length and order of GAD65 and IA2 sequences were critical for conservation of the conformational epitopes in the fusion protein. Among four chimera tested, only IA2((606-979))/GAD65((1-585)) retained wild-type-like folding of GAD65 and IA2 domains and yielded a stable protein after baculovirus expression. Reactivity of GAD65 antibody- and IA2 antibody-positive sera from patients newly diagnosed with insulin-dependent diabetes mellitus, from ICA-positive prediabetics, and from ICA-positive first-degree relatives demonstrated conservation of the relevant autoreactive epitopes. The assay based on the in vitro translated fusion antigen had a sensitivity and specificity identical to those for detection of GAD65- and IA2-abs based on the two separate GAD65 and IA2 proteins. CONCLUSIONS: Autoantigens such as GAD65 and IA2 can be combined successfully in a fusion protein of similar immune reactivity. This allows simultaneous detection of GAD65- and IA2-abs in a one-step screening assay and cost-effective identification of positive individuals at risk of diabetes or at onset of disease.

Animals↗
PubMed2000

Ripple phases induced by alpha-tocopherol in saturated diacylphosphatidylcholines.

The effect of alpha-tocopherol on the structure and phase behavior of dilauroyl-, dimyristoyl-, dipalmitoyl-, and distearoyl-phosphatidylcholines was examined using X-ray diffraction and freeze-fracture electron microscopic methods. A ripple phase was observed in all of the mixtures at temperatures well below the pretransition temperature of the corresponding pure phospholipid. Freeze-fracture studies indicated that with proportion of alpha-tocopherol less than 5 mol% a ripple phase with large periodicity (50-150 nm) predominated and with about 10 mol% alpha-tocopherol a ripple phase of periodicity about 16 nm was formed. With more than 10 mol% alpha-tocopherol planar bilayers tended to be formed. Partial phase diagrams of mixed aqueous dispersions of saturated phosphatidylcholines and alpha-tocopherol over temperature ranges about the gel to liquid-crystal phase boundary have been constructed. Alpha-tocopherol-enriched domains form ripple phases that coexist with regions of lamellar gel phase of the pure phospholipid in mixtures containing less than 10 mol% alpha-tocopherol. The presence of increasing amounts of alpha-tocopherol in the phospholipid causes an increase in the proportion of ripple phase at the expense of pure phospholipid bilayer indicating that the alpha-tocopherol-enriched domains might possess a defined stoichiometry of the two constituents.

Dimyristoylphosphatidylcholine↗
PubMed2000

Resampling as a cluster validation technique in fMRI.

Exploratory, data-driven analysis approaches such as cluster analysis, principal component analysis, independent component analysis, or neural network-based techniques are complementary to hypothesis-led methods. They may be considered as hypothesis generating methods. The representative time courses they produce may be viewed as alternative hypotheses to the null hypothesis, i.e., "no activation." We present here a resampling technique to validate the results of exploratory fuzzy clustering analysis. In this case an alternative hypothesis is represented by a cluster centroid. For both simulated and in vivo functional magnetic resonance imaging data, we show that by permutation-based resampling, statistical significance may be computed for each voxel belonging to a cluster of interest without parametric distributional assumptions.

Brain↗