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Biomedical subjects

J Wouters

Publications and source records attributed to J Wouters.

At least 55 records · Page 3Linked to original sources

Host specificity in the Richelia-diatom symbiosis revealed by hetR gene sequence analysis.

The filamentous heterocyst-forming cyanobacterium Richelia intracellularis forms associations with diatoms and is very abundant in tropical and subtropical seas. The genus Richelia contains only one species, R. intracellularis Schmidt, although it forms associations with several diatom genera and has considerable variation in size and morphology. The genetic diversity, and possible host specificity, within the genus Richelia is unknown. Using primers against hetR, a gene unique for filamentous cyanobacteria, specific polymerase chain reaction (PCR) products were obtained from natural populations of R. intracellularis filaments associated with three diatom genera. Phylogenetic analyses of these sequences showed that they were all in the same clade. This clade contained only the R. intracellularis sequences. The genetic affiliation of hetR sequences of R. intracellularis to those of other heterocystous cyanobacteria strongly suggests that it was not closely related to endosymbiotic Nostoc spp. hetR sequences. Sequences from R. intracellularis-Hemiaulus membranaceus sampled in the Atlantic and Pacific Oceans were almost identical, demonstrating that the genetic relatedness was not dependent on geographical location. All sequences displayed a deep divergence between symbionts from different genera and a high degree of host specificity.

Bacterial Proteins↗

Structure and function prediction of the Brucella abortus P39 protein by comparative modeling with marginal sequence similarities.

A methodology is proposed to solve a difficult modeling problem related to the recently sequenced P39 protein. This sequence shares no similarity with any known 3D structure, but a fold is proposed by several threading tools. The difficulty in aligning the target sequence on one of the proposed template structures is overcome by combining the results of several available prediction methods and by refining a rational consensus between them. In silico validation of the obtained model and a preliminary cross-check with experimental features allow us to state that this borderline prediction is at least reasonable. This model raises relevant hypotheses on the main structural features of the protein and allows the design of site-directed mutations. Knowing the genetic context of the P39 reading frame, we are now able to suggest a function for the P39 protein: it would act as a periplasmic substrate-binding protein.

Amino Acid Sequence↗

Natural vowel and consonant recognition by Laura cochlear implantees.

OBJECTIVE: This study was performed to examine whether spectral and temporal properties of speech sounds are transferred effectively by the Laura cochlear implant device, and to determine whether listeners with different levels of performance use the same type of information to understand speech. DESIGN: Twenty-five adult Laura cochlear implantees identified 12 repetitions of 10 vowels in /hVt/ utterances and 16 consonants placed in intervocalic /a/ context. The stimulus-response confusion matrices were analyzed in terms of relative information transmission scores (Miller & Nicely, 1955), and they were subjected to multidimensional INDSCAL analyses to examine whether a similar pattern of results would emerge when no prior assumptions were made about the division of categories. RESULTS: To examine perception for different levels of performance, the vowel and consonant confusion data were divided into three groups of subjects: the better, intermediate, and poorer performers. In general, the INDSCAL analyses confirmed the results obtained from the information transmission analyses. However, they also supplemented it with other perceptually relevant cues for cochlear implantees, as well as with the weightings over different perceptual dimensions for different types of performers. These analyses suggest that although all subjects use the same type of information, the better performers are more capable of using these different cues than poor performers.

Adolescent↗

Enhancing the speech envelope of continuous interleaved sampling processors for cochlear implants.

A new method to code the speech envelope in continuous interleaved sampling (CIS) processors for cochlear implants is proposed. In this enhanced envelope, the rapid adaptation seen in the response of auditory nerves to sound stimuli is incorporated. Two strategies, one using the standard envelope (CIS) and one using the enhanced envelope (EECIS), were tested perceptually with six postlingually deafened users of the LAURA cochlear implant. The tests included identification of stop consonants in three different vowel contexts and monosyllabic consonant-vowel-consonant (CVC) words. Significant improvements in correct identification scores were observed for stop consonants in intervocalic /a/ context (p = 0.026): average results varied from 46% correct for CIS to 55% for EECIS. This improvement was mainly due to the better transmission of place of articulation. The differences in identification scores for stop consonants in /i/ and /u/ context were not significant. The identification scores for the medial vowels of the CVC words were significantly higher when the EECIS strategy was used: average results increased from 39% correct to 46% correct (p = 0.018). No significant differences were observed between the results for initial and final consonants of the CVC words. The present results demonstrate that the inclusion of the rapid adaptation in the speech processing for cochlear implants can improve speech intelligibility.

Adolescent↗

Gap detection in single- and multiple-channel stimuli by LAURA cochlear implantees.

Gap-detection thresholds were determined for different complex patterns of electrical stimulation in four postlingually deafened LAURA cochlear implantees, to examine the nature of within- and across-channel auditory processes in more detail. Gap detectability was examined as a function of stimulus complexity (one, two, or three channels), channel distance within and across multichannel pre- and post-gap markers, stimulus asymmetry, and pulse rate. All markers roved in duration from 200 to 500 ms to ensure that subjects were not using overall stimulus duration as a cue. Gap-detection thresholds for all subjects were short (< 5 ms) when the pre- and post-gap markers stimulated the same single or multiple channels, even when the distance between simultaneously stimulated channels was large (exp. 1). For some subjects, gap detectability was more difficult in the across-channel condition, when the pre- and post-gap markers each stimulated different channels, although performance improved substantially in most subjects after extensive training (exp. 2). Additional tests with random maskers also suggest that neural interaction only affects performance mildly, and that the magnitude of the gap-detection threshold probably depends more on the subject's cognitive (in)ability to attend to the temporal gap than on the temporal acuity of their auditory system. Other stimulus conditions showed a difference in performance related to the order of the markers: gap thresholds were longer when the pre-gap marker stimulated one channel and the post-gap marker stimulated two or more channels, than vice versa (exp. 3). In addition, gap thresholds of three of the subjects increased with decreasing pulse rate from 1250 to 400 pps, a finding which may be related to the rate of the speech processing strategies used by each individual implantee (exp. 4).

Acoustic Stimulation↗

Design, synthesis, and anticonvulsant activity of 1-(pyrid-3-ylsulfonamido)-2-nitroethylenes.

The lipophilic 1-cycloalkylamino-1-(pyrid-3-yl-sulfonamido)-2-nitr oethylenes were synthesized as bioisosteres of BM-34, an anticonvulsant sulfonylthiourea. Compound 17 (ip) emerged from the maximal electroshock seizure (MES) test with a 50% effective dose (ED50) of 8.25 mg/kg. Its anticonvulsant profile was similar to that of phenytoin (ED50 = 9.51 mg/kg) and of BM-34 (ED50 = 1.19 mg/kg): active in the MES test and inactive in seizures induced by subcutaneous injection of pentetrazole, strychnine, bicuculline, picrotoxin, or N-methyl-D,L-aspartate. The neurotoxicity of 17 (TD50 = 113.8 mg/kg) was lower than that of phenytoin (TD50 = 65.5 mg/kg) but higher than that of BM-34 (TD50 = 147.2 mg/kg). Crystallographic study revealed that BM-401 (17) was a zwitterionic structure. Its sulfonamido nitroethylene side chain adopted a conformation which placed the two cycloalkyl rings face to face to form a single hydrophobic area.

Animals↗

First partial three-dimensional model of human monoamine oxidase A.

A survey of the major known structural aspects of monoamine oxidase (MAO) is given and a first partial model of human MAO A is presented. This 3D model has been established using secondary structure predictions and fold recognition methods. It shows two alpha/beta domains (the FAD-binding N-terminal and central domains) and an alpha+beta domain. The C-terminal region is predicted to be responsible for anchoring the protein into the mitochondrial membrane and was not modeled. The covalent binding of the flavin cofactor to a cysteine residue is well predicted. The model is validated with experimental data from the literature and should be useful in designing new experimental studies (site-directed mutagenesis, chemical modification, specific antibodies). This first step towards the 3D structure of monoamine oxidase should contribute to a better understanding of the mechanisms of action and inhibition of this drug target in the treatment of clinical depression.

Amino Acid Sequence↗

Mutagenesis of the fructose-6-phosphate-binding site in the 2-kinase domain of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase.

Multiple alignment of several isozyme sequences of the bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase revealed conserved residues in the 2-kinase domain. Among these residues, three asparagine residues (Asn76, Asn97 and Asn133; numbering refers to the liver isozyme sequence) and three threonine residues (Thr132, Thr134 and Thr135) are located near the fructose 6-phosphate-binding site in the crystal structure of the bifunctional enzyme. The role of these residues in substrate binding and catalysis in the 6-phosphofructo-2-kinase domain has been studied by mutagenesis to alanine. Since the crystal structure of 6-phosphofructo-2-kinase does not contain fructose 6-phosphate, this substrate was docked into the putative binding site by computer modelling, and its interactions with the protein were predicted. Analysis of the mutagenesis-induced changes in kinetic properties and of the substrate-docking model revealed that all these residues are directly or indirectly involved in fructose-6-phosphate binding. All the mutants displayed an increased Km for fructose 6-phosphate (10-200-fold). We propose that Asn133 stabilises Arg138, which itself makes a direct electrostatic bond with the 6-phosphate group of fructose 6-phosphate, that Asn76 interacts with the C3 hydroxyl group of fructose 6-phosphate, that Thr132 makes a hydrogen bond with the C6 oxygen of this substrate, and that Thr134 interacts with two residues involved in fructose-6-phosphate binding, Thr132 and Tyr199. On the other hand, Asn97 and Thr135 play structural roles, by maintaining the structure of the fructose-6-phosphate-binding pocket.

Base Sequence↗

Molecular lipophilicity potential by CLIP, a reliable tool for the description of the 3D distribution of lipophilicity: application to 3-phenyloxazolidin-2-one, a prototype series of reversible MAOA inhibitors.

The capacity factor of eleven derivatives belonging to a prototype series of 3-phenyloxazolidin-2-one, reversible MAO inhibitors, was measured and compared to the calculated log Pcalc using the CLIP package. We demonstrate that this Molecular Lipophilicity Potential (MLP) approach is a valuable tool to estimate log Pcalc of such compounds.

Chromatography, High Pressure Liquid↗

Cation-pi (Na+-Trp) interactions in the crystal structure of tetragonal lysozyme.

Experimental evidence of a cation-pi interaction between a sodium cation (Na+) and the indole ring of residue Trp123 in a structure (2.0 A) of hen egg-white lysozyme is presented. The geometry of the metal ion-pi interaction observed in the protein structure (distance between the aromatic plane and the cation approximately 4 A) is consistent with geometries observed among small molecules crystal structures and quantum chemistry ab initio calculations. The present crystal structure of lysozyme provides unique structural information about the geometry of binding of cations to pi systems in proteins. It shows that the metal ion-pi interaction within proteins is not significantly different from similar bindings found in small molecules and that it can be modeled by theoretical methods.

Binding Sites↗

An adaptive noise canceller for hearing aids using two nearby microphones.

An adaptive noise cancellation scheme based on two-stage adaptive filtering as proposed by Van Compernolle [Proceedings of the IEEE International Conference on Acoustics, Speech and Signal Processing (IEEE, New York, 1990)] was tested for two identical directional microphones mounted in an endfire configuration within a single behind-the-ear (BTE) hearing aid. Speech intelligibility tests were carried out with two normal hearing subjects and three hearing aid users with moderate hearing losses, in a realistic test environment using open set words and sentences. A significant improvement, on average more than 5 dB, of speech reception threshold (SRT) in background noise was obtained.

Amplifiers, Electronic↗

Structural aspects of monoamine oxidase and its reversible inhibition.

Among the various types of amine oxidases, monoamine oxidase (MAO) is of special interest for neuropsychiatry. It appears to play a central role in several psychiatric and neurological disorders, including clinical depression and Parkinson's Disease. The purpose of this paper is to review the major structural aspects of MAO, in relation with its primary sequence (site-directed mutagenesis, chimeric constructions), secondary and three-dimensional structures. The consequences of the inhibition of monoamine oxidase are considered, mainly focusing on the new generations of reversible and selective MAO inhibitors. Recent efforts to design and develop those inhibitors are emphasized and discussed.

Amino Acid Sequence↗

Secondary structure of monoamine oxidase by FTIR spectroscopy.

The secondary structure of human monoamine oxidase A and bovine monoamine oxidase B has been investigated by Fourier Transform Attenuated Total Reflection Spectroscopy (FTIR ATR). The experimental results are compared for both isoenzymes and the data are incorporated in a statistical attribution of secondary structure of the enzyme describing the distinct folding and molecular specificity of the two types of monoamine oxidase.

Amino Acid Sequence↗

Experimental and theoretical study of reversible monoamine oxidase inhibitors: structural approach of the active site of the enzyme.

Experimental and theoretical physico-chemical methods were used to investigate the interaction between aryl-oxazolidinones and monoamine oxidase (MAO). Several arguments suggest that these compounds interact with the flavin adenine dinucleotide (FAD) cofactor of MAO. The calculation using ab initio molecular orbital methods of the electronic properties of flavin and befloxatone, a reversible inhibitor of MAO A, led to a description of the interaction between aryl-oxazolidinones and the cofactor of the enzyme. Structure activity relationship results revealed additional sites of interaction with the protein core of MAO A. As a result of this work, a model is proposed for the reversible inhibition of MAO by oxazolidinones via long distance, reversible interactions with the FAD cofactor of the enzyme.

Chemical Phenomena↗

Acoustic analysis of tracheo-oesophageal versus oesophageal speech.

In order to evaluate the vocal quality of tracheo-oesophageal and oesophageal speech, several objective acoustic parameters were measured in the acoustic waveform (fundamental frequency, waveform perturbation) and in the frequency spectrum (harmonic prominence, spectral slope). Twelve patients using tracheo-oesophageal speech (with the Provox valve) and 12 patients using oesophageal speech for at least two months, participated. The main results were that tracheo-oesophageal voices more often showed a detectable fundamental frequency, and that this fundamental frequency was fairly stable; there was also a tendency to more clearly defined harmonics in tracheo-oesophageal speech. This suggests a more regular vibratory pattern in the pharyngo-oesophageal segment, due to the more efficient respiratory drive in tracheo-oesophageal speech. So, a better quality of the voice can be expected, in addition to the longer phonation time and higher maximal intensity.

Evaluation Studies as Topic↗

Kinetic characterization of tyramine oxidase of Arthrobacter species.

Amine oxidases (EC 1.4.3) represent a rather nebulous group of proteins with relative narrow substrate and inhibitor specificities. Several different enzymes fall into these amine oxidase classes and the classification of some of them still remains ambiguous. Kinetic and physico-chemical properties of tyramine oxidase of Arthrobacter sp. were investigated to decide if this enzyme belongs to the flavin containing tyramine oxidase class (EC 1.4.3.9) or if it is more related to another amine oxidase class. On the basis of its spectral characteristics, molecular weight and inhibition profile, the enzyme was identified as a semicarbazide sensitive copper-containing amine oxidase.

Arthrobacter↗