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R Thalmann

Publications and source records attributed to R Thalmann.

At least 37 records · Page 2Linked to original sources
PubMed1994

Protein profile of human perilymph: in search of markers for the diagnosis of perilymph fistula and other inner ear disease.

Recent developments in high-resolution two-dimensional polyacrylamide gel electrophoresis, combined with amino acid sequencing and computer-assisted image analysis, have allowed separation of approximately 100 proteins and identification and quantitation of some 30 proteins in human perilymph. The majority of proteins were found to be present in perilymph at levels in basic agreement with the total protein gradient between perilymph and plasma (1:35). However, several striking differences were observed: (1) beta 2-transferrin, known to be absent from normal plasma but present in cerebrospinal fluid, was detected in perilymph at a concentration roughly equal to that in cerebrospinal fluid; and (2) two high-density lipoprotein-associated apolipoproteins--apo D (formerly PLS:33) and apo J or NA1 and NA2 (formerly PSL:29/30), the latter showing identity with SP40/40, or cytolysis inhibitor--were found to be present at concentrations 1 to 2 orders of magnitude higher when examined in terms of total protein and to be comparable with or higher than plasma levels when examined in terms of absolute concentrations. The functional significance of the extremely high levels of the two apolipoproteins is not known at this time. An attempt was made to use beta 2-transferrin, as well as apo D and apo J (NA1/NA2), as markers for the diagnosis of perilymph fistula, one of the most controversial and challenging problems for the otologist today. It was determined that the technique is indeed applicable when relatively pure fistula samples are analyzed. Limitations and potential improvements of the technique are discussed. In addition, the potential usefulness of two-dimensional polyacrylamide gel electrophoresis in other pathologic conditions of the inner ear is discussed briefly.

Apolipoproteins↗
PubMed1993

Uronic acid-containing glycosaminoglycans and keratan sulfate are present in the tectorial membrane of the inner ear: functional implications.

The tectorial membrane is a gel-like, acellular connective tissue overlying the microscopic organ of Corti--the auditory sensory structure. It is instrumental in the sound-synchronous deflection of the stereocilia of the hair cells, a central event in auditory transduction. It is well established that collagen, primarily type II, constitutes the major protein of the tectorial membrane, with smaller amounts of types IX and XI also present. However, conclusive information on the proteoglycans in this structure is lacking. Tectorial membranes were extracted with a 4 M guanidine--HCl solvent, and proteoglycans isolated after ethanol precipitation and collagenase treatment. A colorimetric assay based on the binding of the cationic dye safranin O to glycosaminoglycans, in combination with enzymatic techniques, detected significant amounts of chondroitin sulfate and keratan sulfate (0.29 and 0.17% on a wet weight basis, respectively). Agarose-polyacrylamide electrophoresis of chondroitinase-digested samples revealed a core protein with a similar molecular mass to that of the large cartilage proteoglycan aggrecan. This proteoglycan reacted with the antibody 3-B-3 (recognizing modified chondroitin 6-sulfate linkage region oligosaccharides). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed several low molecular mass proteins which reacted with 5-D-4, specific for keratan sulfate, one of which showed characteristics of fibromodulin. Comparison of the quantitative aspects of various connective tissue components of tectorial membrane with other type II collagen-containing structures revealed that this tissue resembles highly hydrated cartilage.

Animals↗
PubMed1993

Partial amino acid sequences of organ of Corti proteins OCP1 and OCP2: a progress report.

Progress in amino acid sequencing of two low-molecular weight acidic proteins of unknown function which are present at high concentrations in the organ of Corti is reported. These two proteins, provisionally termed OCP1 and OCP2, were originally demonstrated by two-dimensional polyacrylamide gel electrophoresis; their presence at high concentrations in the inner ear sensory epithelia strongly suggests that OCP1 and OCP2 serve some important function in the ear. In the present paper, extension of the amino-terminal sequence of OCP2 to 71 residues is described. In addition, the first results on sequencing of OCP1 are presented. Computer algorithms are used to predict important structural features, and the sequences are analyzed for probable phosphorylation, glycosylation, and Ca-binding sites. Projected further studies in immunochemistry and molecular biology of OCP1 and OCP2 based on the amino acid sequencing data are discussed.

Amino Acid Sequence↗
PubMed1993

Differential physiologic effects of perfusion of scala tympani versus scala vestibuli in the ischemic cochlea.

The effectiveness of perilymphatic perfusion with oxygenated artificial media upon the endocochlear potential (EP) was measured during systemic ischemia in the guinea pig. Differences in the effects of perfusion of the two perilymphatic scalae were determined. Perfusion of scala vestibuli with oxygenated artificial perilymph at a high flow rate resulted in complete recovery of the EP to the pre-ischemic level, whereas perfusion of scala tympani with the same medium was unable to effect complete recovery. The recovery obtained by perfusion of scala tympani was about half that obtained of scala vestibuli. The pO2 in scala media was measured during perfusion by means of oxygen-sensitive microelectrodes. perfusion of scala vestibuli led to an approximately two-fold higher pO2 in scala media than perfusion of scala tympani. During perfusion, the pO2 in scala media varied dependent upon depth of electrode insertion, with a gradient decreasing toward the stria vascularis, a direction opposite to that seen under normal metabolic conditions. These findings suggest that, in the ischemic cochlea, oxygen enters scala media more easily from scala vestibuli across Reissner's membrane than from scala tympani via the basilar membrane/organ of Corti complex.

Animals↗
PubMed1993

[Biochemical profile of proteoglycans and glucosaminoglycans of the mammalian tectorial membrane].

The tectorial membrane is an acellular connective tissue which plays an essential role in cochlear function. While a comparatively large amount of information is available on the collagen network of the tectorial membrane, studies on the biochemical nature of this highly hydrated matrix, which is composed of proteoglycans (PGs) and glycosaminoglycans (GAGs), have been quite limited. Previous reports on the biochemical analysis of the tectorial membrane have failed to detect uronic acid, which is present in large amounts in all mammalian GAGs except keratan sulfate. Applying a colorimetric assay based on the binding of GAGs to cationic dye Safranin-0 in combination with enzymatic techniques, we were able to measure GAGs in the murine tectorial membrane. Approximately 0.3% uronic acid-containing GAGs (mainly in the form of chondroitin/dermatan sulfate) and 0.17% keratan sulfate were detected in the tectorial membrane (both on a wet weight basis). In addition, various types of electrophoresis revealed one large PG with a molecular mass similar to that of the large type cartilage PGs and three small PGs, containing chondroitin sulfate and keratan sulfate side chains, respectively. Judging by coelution of standards, one of the small PGs seemed to correspond to fibromodulin, which has at least one keratan sulfate side chain, and binds to type I and type II collagen to regulate collagen organization in tissues. Our results suggest: (1) Donnan equilibrium is established in the tectorial membrane because sulfated GAGs are highly negatively charged and consequently bring about an influx of large amounts of water and cations into the matrix.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Protein profiles of perilymph and endolymph of the guinea pig.

Results of protein separation of guinea pig plasma, perilymph, and endolymph by means of high-resolution two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis are presented. Several proteins are present in perilymph at levels in basic accord with the total protein gradient with respect to plasma; however, others are present in perilymph at levels comparable to plasma levels, and one protein low molecular weight protein, PLS:33, is eight times higher. In addition, a high molecular weight protein is shown to be present at similar levels in the two compartments. These findings indicate that ultrafiltration cannot be the sole mechanism of perilymph production. Endolymph proteins are uniformly five to eight times lower than perilymph levels, essentially following the total protein concentration gradient between the two compartments. This supports the view that endolymph is derived from perilymph rather than directly from blood.

Animals↗
PubMed1991

Radial communication between the perilymphatic scalae of the cochlea. I: Estimation by tracer perfusion.

The degree to which radial exchange between scala tympani (ST) and scala vestibuli (SV) can occur has not previously been quantified. We have measured the amount of cross-communication in the third turn of the guinea pig cochlea using an ionic tracer, trimethylphenylammonium (TMPA). TMPA was perfused through one scala while TMPA concentrations were measured simultaneously in the perfused and non-perfused scalae. On the basis of the time course of TMPA increase recorded in the non-perfused scala, we were able to calculate rate constants for cross-leak and clearance. Cross-leak in the third turn occurred remarkably rapidly with a rate constant from ST to SV of 0.049 min-1 and from SV to ST of 0.031 min-1. These correspond to transfer half times of 14.2 and 22.4 min respectively. This result demonstrates that ST and SV in the third turn cannot be regarded as independent compartments.

Animals↗
PubMed1991

Radial communication between the perilymphatic scalae of the cochlea. II: Estimation by bolus injection of tracer into the sealed cochlea.

Radial communication between ST and SV was measured in the sealed cochlea by monitoring the dispersal of an ionic tracer, trimethylphenylammonium (TMPA) injected in the form of a minute bolus. Tracer movements were recorded by a pair of ion-selective electrodes sealed into the injected and non-injected scalae close to the injection site. Measurements were made in the basal or third turn of the guinea pig cochlea. In the third turn, radial communication occurred rapidly with a ST half time from ST to SV of 25 min and from SV to ST of 26 min. In the basal turn the communication was markedly slower, with a ST half time from ST to SV of 170 min and from SV to ST of 240 min. However, the difference between the basal and third turns can be shown to arise almost totally from differences in cross-sectional area of the perilymphatic scalae. When normalized with respect to scala cross-section, the process of tracer movement across the spiral ligament is similar in the basal and third turns. These results demonstrate that radial communication between scala tympani and scala vestibuli is an important route which must be considered in studies involving perilymph.

Animals↗
PubMed1991

Evaluation of procedures to reduce fluid flow in the fistulized guinea-pig cochlea.

The rate of longitudinal flow of fluid in scala tympani (ST) has been quantified under a number of experimental conditions. The method used to measure flow involved using a tracer ion (trimethylphenylammonium: TMPA) as a volume flow marker. Movement of marked perilymph was monitored by ion-selective microelectrodes which were capable of detecting exceedingly low concentrations of TMPA. Our results show that when the cochlea is perforated at the apex, flow rates of 400-500 nl/min are induced in ST, compared to the normal very slow rate of 2 nl/min when the cochlea is sealed. This artifactual flow of CSF through the perforated cochlea can be reduced to 6.9 nl/min by releasing the hydrostatic pressure of cerebrospinal fluid (CSF) or further reduced to 1.8 nl/min by surgically obstructing the cochlear aqueduct. In addition, we observed no basally-directed flow in ST when the round window (RW) was perforated, demonstrating that perilymph is not produced in volume as previously assumed. This study demonstrates the importance of separating artifactual flows, induced by the experimental procedures required to access the cochlear fluids, from the low flow rates which occur in normal, physiologic conditions.

Action Potentials↗
PubMed1991

[Coronary endoprosthesis. Short- and mid-term results].

Mechanical stenting of the coronary wall by an endoprosthesis is a method of treating and preventing the 2 major complications of balloon angioplasty: acute occlusion by dissection and secondary restenosis. Forty-four self expanding stents were implanted in 42 patients presenting with post-angioplasty restenosis (35 cases), stenosis of an aortocoronary bypass graft (4 cases) or symptomatic post-angioplasty coronary dissection (3 cases). The primary results featured a high early occlusion rate of the endoprosthesis (28.5%, 12 cases). Better patient selection to include patients with stable angina and coronary arteries with a diameter greater than 3 millimeters, and the administration of an association of platelet antiaggregant therapy with heparin and oral anticoagulants has led to an improvement in these results. The early occlusion rate in the first 23 patients implanted without these selection criteria was 43% (10 cases) whereas it was only 10.5% (2 cases) in the last 19 selected patients who were prescribed effective medical therapy. The medium term results of the 27 patients implanted without complications at 6 months showed restenosis in 5 cases and reocclusion (asymptomatic) in 1 case (22.2% at control angiography at the 6th month). Restenosis was only observed in cases in which the endoprosthesis did not completely cover the coronary lesion (3 out of 3 cases) or when the endoprosthesis was implanted in a saphenous vein graft (2 out of 4 cases) and the only case of reocclusion occurred on a stent implanted in a coronary artery responsible for previous myocardial infarction. In the 21 patients in whom the stenosis had been totally covered by the stent there were no cases of restenosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1990

Partial amino acid sequence of organ of Corti protein OCP-II.

We have previously described two low-molecular-weight, highly acidic proteins which are present in the organ of Corti in extremely high concentrations. Since the function of these proteins is not known, they have been assigned the tentative names OCP-I and OCP-II. In the hope of obtaining information about their function through homology studies, we have initiated amino acid sequencing of these proteins. We have recently succeeded in obtaining a brief amino-terminal sequence of OCP-II. We now report on a significant extension of the amino-terminal sequence of OCP-II and our first results on sequences of peptide fragments obtained by limited digestion with V8 protease. Together, the sequenced segments account for about one-third of the total sequence. Comparisons with the sequences of known proteins suggest that OCP-II is not a structural protein, but that it may exhibit biologic activity.

Amino Acid Sequence↗
PubMed1990

Biochemical features of major organ of Corti proteins (OCP-I and OCP-II) including partial amino acid sequence.

Further biochemical and biophysical characterization of two low-molecular-weight, strongly acidic proteins that are present at extremely high levels in the organ of Corti, tentatively named OCP-I and OCP-II, is presented. The two proteins are also present, although at much lower levels, in the vestibular end-organs and a variety of other inner ear tissues; they have not been observed in other systems. OCP-I and II are highly soluble and do not contain appreciable amounts of carbohydrate. The two proteins, originally described in the guinea pig, are compared electrophoretically with the corresponding proteins in several other mammalian species. Preliminary data on the amino acid composition of the two proteins are presented. Moreover, the amino-terminal sequence of a 22-residue segment of OCP-II is shown and compared to the sequences of known proteins.

Amino Acid Sequence↗
PubMed1989

Perilymph composition in scala tympani of the cochlea: influence of cerebrospinal fluid.

A commonly used technique to obtain cochlear perilymph for analysis has been the aspiration of samples through the round window membrane. The present study has investigated the influence of the volume withdrawn on sample composition in the guinea pig. Samples of less than 200 nl in volume taken through the round window showed relatively high glycine content, comparable to the level found in samples taken from scala vestibuli. If larger volumes are withdrawn, lower glycine levels are observed. This is consistent with cerebrospinal fluid (having a low glycine content) being drawn into scala tympani through the cochlear aqueduct and contaminating the sample. The existence of a concentration difference for glycine between scala tympani perilymph and cerebrospinal fluid suggests the physiologic communication across the cochlear aqueduct is relatively small in this species. The observation of considerable exchange between cerebrospinal fluid and perilymph, as reported in some studies, is more likely to be an artifact of the experimental procedures, rather than of physiologic significance. Alternative sampling procedures have been evaluated which allow larger volumes of uncontaminated scala tympani perilymph to be collected.

Animals↗
PubMed1989

[Vascular endoprosthesis of femoro-popliteal occlusive disease].

A self-expanding vascular endoprosthesis (wall stent) was implanted in 26 patients with femoropopliteal occlusive disease following recurrent stenosis or occlusion after percutaneous angioplasty. Implantation was successful in all cases and there were no complications. Five of the 26 patients developed a thrombosis in the first nine days; in four of these, thrombolysis was successful. Patency rate after one month was 96%, after three months 95%, after six months 85% and after nine months 87%. In order to prevent thrombosis after implantation, anticoagulant therapy is indicated. Poor distal flow encourages intimal hyperplasia and therefore recurrences.

Adult↗
PubMed1988

Volume flow rate of perilymph in the guinea-pig cochlea.

The rate of longitudinal flow of perilymph has been measured using an ionic tracer technique. Spread of the tracer trimethylphenylammonium (TMPA) along the perilymphatic scalae was monitored with ion-selective microelectrodes following injection of a minute bolus (approximately 50 nl) of 150 mM TMPAC1 one turn away. This amount of TMPA had virtually no toxic effect on cochlear function. The spread of tracer by longitudinal volume flow and passive diffusion were separated by comparing tracer movements in both apical and basal directions along the scalae in two groups of animals. Experimental findings were compared with a mathematical model which combined diffusion and volume flow. Our results demonstrated that when electrodes were completely sealed into the cochlea, the rate of longitudinal volume flow in scala tympani was extremely slow, approximately 1.6 nl/min in the apical direction. Longitudinal flow was not detectable in scala vestibuli. When the otic capsule was perforated, flow rates of over 1 microliter/min were recorded in scala tympani, probably as a result of cerebrospinal fluid entry through the cochlear aqueduct. When the cochlea was sealed (with recording electrodes in place) and cerebrospinal fluid pressure was released, there was no significant basally-directed flow of perilymph in scala tympani. These findings support the concept that perilymph composition is maintained by local, cochlear mechanisms which do not involve longitudinal volume flow. They provide strong evidence that perilymph is not secreted in one region and resorbed at a spatially distant site.

Action Potentials↗
PubMed1988

Melatonin and other serotonin derivatives in the guinea pig membranous cochlea.

In mammals, the indolealkylamine melatonin is synthesized mainly in the pineal gland and to a lesser degree in a number of extrapineal sites. To obtain more information on its distribution, melatonin was measured in the membranous cochlea of the guinea pig. Moreover, melatonin and other serotonin derivatives were determined in organ cultures after incubation with 14C-labelled serotonin. The results show that melatonin is detectable in the organ of Corti and the basilar membrane, and to a lesser degree also in the cochlear nerve and stria vascularis, including the spiral ligament. In vitro studies reveal the occurrence of a number of radiolabelled serotonin derivatives indicating that two of the melatonin-forming enzymes, serotonin N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT) are present in membranous cochlea. It is concluded that melatonin is synthesized in the guinea pig cochlea. Its role as modulator of hair cell function is dissolved.

Acetylserotonin O-Methyltransferase↗