Surgical management of an epithelioid hemangioendothelioma of the superior vena cava protruding into the right atrium.

Epithelioid hemangioendothelioma (EH) is a rare malignant tumor of vascular origin that often involves soft tissues and visceral organs, and less commonly, large veins. We report a case of EH of the superior vena cava protruding into the right atrium and its surgical management.

Calculation and analysis of the harmonic vibrational frequencies in molecules at extreme pressure: methodology and diborane as a test case.

We present a new quantum chemical method for the calculation of the equilibrium geometry and the harmonic vibrational frequencies of molecular systems in dense medium at high pressures (of the order of GPa). The new computational method, named PCM-XP, is based on the polarizable continuum model (PCM), amply used for the study of the solvent effects at standard condition of pressure, and it is accompanied by a new method of analysis for the interpretation of the mechanisms underpinning the effects of pressure on the molecular geometries and the harmonic vibrational frequencies. The PCM-XP has been applied at the density functional theory level to diborane as a molecular system under high pressure. The computed harmonic vibrational frequencies as a function of the pressure have shown a satisfactory agreement with the corresponding experimental results, and the parallel application of the method of analysis has reveled that the effects of the pressure on the equilibrium geometry can be interpreted in terms of direct effects on the electronic charge distribution of the molecular solutes, and that the effects on the harmonic vibrational frequencies can be described in terms of two physically distinct effects of the pressure (curvature and relaxation) on the potential energy for the motion of the nuclei.

Patient-specific and real-time model of numerical simulation of the hemodynamics of type B aortic dissections.

INTRODUCTION: Regular monitoring of uncomplicated type B aortic dissection is essential because 25-30% will progress to aneurysmal form. The predictive factors of this evolution are not clearly defined, but they seem to be correlated with hemodynamic data. HYPOTHESIS: Our goal is to create a patient-specific and real-time model of numerical simulation of the hemodynamics of uncomplicated type B aortic dissections in order to predict the evolution of these pathologies for earlier treatment. METHOD: This model consists in a coupling 0D (hydraulic-electric analogy) - 3D (CT angiography segmentation) of the aortic arch with optimization by comparison to the 2D Phase Contrast MRI data and using Reduced Order Models to drastically reduce computing times. We tested our model on a healthy and a dissected patient. Then we realized different systolic blood pressure scenarios for each case, which we compared. RESULTS: In the dissected patient, the blood pressure at the false lumen wall was less important than the true lumen. Furthermore, the aortic wall shear stress and the velocity fields in aorta increase at the entry and re-entry tears between the two lumens. The simulation of different blood pressures scenarios shows a decrease in all these three parameters related to the decrease of the systolic blood pressure. CONCLUSION: Our model provides reliable patient-specific and real-time 3D rendering. It has also allowed us to realize different flow variation scenarios to simulate different clinical conditions and to compare them. However, the model still needs improvement in view of a daily clinical application.

Expression level-dependent contribution of glucocorticoid receptor domains for functional interaction with STAT5.

The action of the glucocorticoid receptor (GR) on beta-casein gene transcription serves as a well-studied example of a case where the action of the GR is dependent on the activity of another transcription factor, STAT5. We have investigated the domain-requirement of the GR for this synergistic response in transfection experiments employing GR mutants and CV-1 or COS-7 cells. The results were influenced by the expression levels of the GR constructs. At low expression, STAT5-dependent transactivation by mutants of the GR DNA binding domain or N-terminal transactivation domain was impaired and the antiglucocorticoid RU486 exhibited a weak agonistic activity. When the N-terminal region of the GR was exchanged with the respective domain of the progesterone receptor, STAT5-dependent transactivation was reduced at low and high expression levels. Only at high expression levels did the GR exhibit the properties of a coactivator and enhanced STAT5 activity in the absence of a functional DNA binding domain and of GR binding sites in the proximal region of the beta-casein gene promoter. Furthermore, at high GR expression levels RU486 was nearly as efficient as dexamethasone in activating transcription via the STAT5 dependent beta-casein gene promoter. The results reconcile the controversial issue regarding the DNA binding-independent action of the GR together with STAT5 and provide evidence that the mode of action of the GR depends not only on the type of the particular promoter at which it acts but also on the concentration of the GR. GR DNA binding function appears to be mandatory for beta-casein gene expression in mammary epithelial cells, since the promoter function is completely dependent on the integrity of GR binding sites in the promoter.

[Pathophysiology of cholesteatoma]. / La physiopathogénie des cholestéatomes.

OBJECTIVE: To describe the development of cholesteatoma using current knowledge. METHOD: Review of the literature. RESULTS: Cholesteatoma describes a mass of keratin (skin) in the middle ear which consists of a perimatrix and matrix. There are at least three kinds of cholesteatoma in the middle ear one resulting from invagination (retraction's pocket), another from migration and the last one from congenital inclusion. Cholesteatoma needs three successive inflammatory phases, the first leading to a retraction pocket, the second leading to pathology of the epidermis and of the floor of the external auditory canal and the third is the actual phase of cholesteatoma with invasion and middle ear auto-destruction with bone resorption. In this last phase, many factors play a role, collagenasis, osteoclats, cytokines, NO, bacteria and their biofilm and rupture of the retraction pocket. CONCLUSION: Cholesteatoma is an inflammatory disease of the ear caracterised by bone resorption. Current research is starting to appreciate the important role the immune system plays in the pathophysiology of cholesteatoma.

Amplitude regulation of episodic release, in vitro biological to immunological ratio, and median charge of human chorionic gonadotropin in pregnancy.

In the present study, we examined the regulation of 24-h serum immunoreactive levels, in vitro biological to immunological (B/I) ratio, and median charge of circulating CG at the end of the first, second, and third trimesters of human gestation. Seven pregnant women were prospectively studied at 12-15, 23-26, and 35-38 weeks of gestation. Blood was sampled every 20 min over a 24-h period, and serum CG concentrations were determined by RIA. Pulse detection and analysis of the 24-h rhythm of serum immunoreactive CG concentrations were carried out by the program Cluster and cosine curve fitting, respectively. The in vitro biological activity of circulating CG was determined by the mouse Leydig cell-testosterone production bioassay, and the median charge of its isoforms was determined by zone electrophoresis in agarose suspension. The immunoreactive levels of CG present at the end of each trimester of gestation fluctuated over a 24-h period; such variability exceeded that of the within-assay coefficient of variation of the CG RIA and could be resolved into a series of CG peaks and valleys. Although no trend in the number of peaks or valleys was systematically found in relation to gestational age, comparisons between the amplitude and area of the CG peaks revealed that these pulse parameters were significantly higher at 12-15 weeks than at 23-26 and 35-38 weeks of gestation. Cosine fits for 24-h rhythms revealed the existence of significant nyctohemeral profiles of serum CG levels in all women studied at 12-15 weeks, in four subjects at 23-26 weeks, and in six women at 35-38 weeks gestation. The time of acrophase was highly homogeneous only between 12-15 weeks of gestation, occurring between 1057-1452 h in six of the women. The in vitro B/I ratio of CG contained in serum pools from 12-15 weeks was significantly (P < 0.05) higher than that exhibited by CG during later gestational periods (B/I ratio at the end of first trimester, 1.14 +/- 0.14; second trimester, 0.87 +/- 0.22; third trimester, 0.79 +/- 0.12). hCG isoforms at 12-15 weeks were more negatively charged than those circulating at 23-26 and 35-38 weeks of gestation. There were no significant differences between the B/I ratio and the median charge of CG molecules from the second and third trimesters. We conclude that serial serum concentrations of CG throughout pregnancy show significant amplitude-modulated pulsatile release and nyctohemeral variations.(ABSTRACT TRUNCATED AT 400 WORDS)

Use of whole Streptococcus pneumoniae cells as a solid phase sorbent for C-reactive protein measurement by ELISA.

Monolayers of pneumococcus (serotype 27) on flat bottom polystyrene microtiter plates were used as a solid phase sorbent for the determination of C-reactive protein (CRP) by ELISA. After binding to the monolayer, CRP was quantified with peroxidase conjugated rabbit anti-human CRP immunoglobulin. The method is sensitive (5 micrograms/ml), rapid (less than 2 h) and correlates well with a laser nephelometric assay (r = 0.95, P less than 0.001), and with a classical sandwich ELISA (r = 0.95, P less than 0.001).

ELISA using whole Legionella pneumophila cell as antigen. Comparison between monovalent and polyvalent antigens for the serodiagnosis of human legionellosis.

An indirect enzyme-linked immunosorbent assay (ELISA) using the six serogroups of whole L. pneumophila bound to microtitre plate wells is described for the serodiagnosis of legionellosis. Comparative studies using monovalent antigen indicated a high correlation between ELISA and indirect immunofluorescence antibody (IFA) tests (r = 0.90, P less than 0.001). Testing 196 human sera by ELISA using both monovalent and polyvalent antigens have established the efficiency of the polyvalent antigen for screening purposes. The ELISA test system exhibited rapidity, sensitivity and reproducibility and should be considered as an alternative to the IFA test for routine serodiagnosis of legionellosis.

Differential effects of the charge variants of human follicle-stimulating hormone.

FSH is synthesized and secreted by the anterior pituitary gland in multiple molecular forms; the release of these isoforms depends on the endocrine status of the donor at the time of sample collection. In the present study, we analysed the possibility that the FSH charge isoforms may exert differential effects at the target cell. Seven FSH isoform mixes were isolated from pooled anterior pituitary glycoprotein extracts by high resolution chromatofocusing, followed by affinity chromatography, which removed nearly 90% of the LH that co-eluted with the FSH isoforms during chromatofocusing. The isoforms (isoform I, pH >7.10; II, pH range 6.60-6.20; III, pH 5. 47-5.10; IV, pH 5.03-4.60; V, pH 4.76-4.12; VI, pH 4.05-3.82 and VII, pH <3.80) were then tested for their capacity to stimulate cAMP release, androgen aromatization and tissue-type plasminogen activator (tPA) enzyme activity and cytochrome P450 aromatase, tPA and inhibin alpha-subunit mRNA production by rat granulosa cells in culture. cAMP and oestradiol production were determined by RIA, tPA enzyme activity by SDS-PAGE and zymography and all mRNAs by northern blot hybridization analysis and semiquantitative RT-PCR. All isoforms, with the exception of isoform I, stimulated synthesis and release of cAMP, oestrogen and tPA enzyme activity in a dose-dependent manner; the potency of the less acidic isoforms (pH 6. 60-4.60) was greater than that exhibited by the more acidic/sialylated analogs (pH 4.76 to <3.80; potencies II>III>IV>V>VII>VI). A similar trend was observed in terms of cytochrome P450 aromatase and tPA mRNA production. In contrast, when FSH-stimulated production of alpha-inhibin mRNA was analysed, isoforms V-VII were significantly more potent (two- to threefold) than the less acidic/sialylated counterparts (II-IV). In contrast to isoforms II-VII (which behaved as FSH agonists), isoform I (elution pH >7.10) completely blocked P450 aromatase and tPA mRNA expression, without altering that of a constitutively expressed gene (glyceraldehyde-3-phosphate dehydrogenase). These results show for the first time that the naturally occurring human FSH isoforms may exhibit differential or even unique effects at the target cell level.

Assessment of the in vitro and in vivo biological activities of the human follicle-stimulating isohormones.

Gonadotropins are synthesized and released in different molecular forms. In this article, we present evidence that the glycosylation variants of human pituitary FSH exhibit differential and divergent effects at the target cell level and that less sialylated, short-lived variants may exert significant effects in in vivo conditions. Less acidic/sialylated glycoforms (elution pH value 6.60-4.60 as disclosed by high resolution chromatofocusing of anterior glycoprotein extracts), induced higher cAMP release, estrogen production and tissue-type plasminogen activator (tPA) enzyme activity as well as cytochrome P450 aromatase and tPA mRNA expression in cultured rat granulosa cells than the more acidic analogs (pH<4.76). By contrast, the more acidic/sialylated glycoforms induced higher alpha-inhibin subunit mRNA expression than their less acidic counterparts. In cumulus enclosed oocytes isolated from mice ovaries, addition of less acidic isoforms induced resumption of meiosis more efficiently than the more acidic analogs. Interestingly, the least acidic isoform (pH>7.10) behave as a strong antagonist of several FSH-mediated effects. Assessment of the in vivo effects of the isoforms on granulosa cell proliferation in follicles from immature rats, revealed that short-lived isoforms were equally or even more efficient than their more acidic counterparts in maintaining granulosa cell proliferation when administered immediately after hypophysectomy. These results show that the naturally occurring human FSH isoforms may exhibit differential or even unique effects at the target cell level and that factors other than the metabolic clearance rate of the molecule (including receptor-binding affinity and capability of the ligand to activate its receptor and trigger intracellular signaling) also play an important role in determining the net in vivo effects of a particular FSH variant.

Preformed antibody and complement rebound after plasma exchange: analysis of immunoglobulin isotypes and effect of splenectomy.

UNLABELLED: Splenectomy (Sx) has been proposed to attenuate post-PE (plasma exchange) rebound of isoagglutinins and xenogenic (XG) antibody (Ab) in both ABO-incompatible allografts and discordant xenografts. This study analyses the qualitative nature and kinetics of serum immunoglobulins as well as complement resynthesis after PE in sham-operated (PE) and splenectomized (PE+Sx) syngeneic LOU/C rats; non-PE sham-operated or splenectomized animals were used as controls. PE was performed in unanesthetized, unheparinized rats. Immunoglobulin isotypes and subclasses (IgM, IgG1, IgG2 alpha, IgG2b) of total circulating Ab were measured pre-PE and up to 21 days post-PE, using ELISA (enzyme-linked immunosorbent assay) and specific mouse antirat monoclonal Ab. Antiguinea-pig (GP) XG Ab (IgM, IgG2a) serum levels were measured using cellular ELISA with cultured GP endothelial cells as targets. Sx alone significantly reduced XG IgM serum levels (p < 0.0001). Maximal rebound of total and XG IgM was observed on day 3 post-PE, reaching 674% and 187% of the pre-PE levels, respectively; these overshoots were entirely suppressed by Sx (p < 0.005 for total IgM; p < 0.0001 for XG IgM). Total IgG2a, IgG2b and IgG1 as well as XG IgG2a serum levels did not show significant overshoot post-PE. The activity of the complement classical pathway (mean +/- SD), assessed by CH50, was decreased at 51 +/- 19% of basal value 15 minutes after PE, and had returned to baseline level by day 2 post-PE with or without Sx. IN CONCLUSION: (1) Six alone significantly reduced XG IgM serum levels; (2) early post-PE Ab rebound was mainly observed for IgM; (3) both total and XG IgM rebound was inhibited by Sx. This suggests that Sx probably removes a significant proportion of IgM producing cells undergoing post-PE stimulation. These data provide a rationale for combining PE with Sx in ABO-incompatible and discordant XG transplantation.

Clinical and serological studies in a series of 45 patients with Guillain-Barré syndrome.

We retrospectively reviewed the clinical files of 45 Guillain-Barré syndrome (GBS) patients admitted to our Department between 1979 and 1989. The age distribution was bimodal with a first peak in young adults (20-40 years), and a second one between 60 to 70 years. Seasonal distribution showed a late fall and a hivernal predominance. Three patients experienced a second attack of GBS 2-9 years after the first one. Thirty-one (69%) presented antecedent events, most often a respiratory tract infection (n = 20) or enteritis (n = 6). Serological studies were systematically performed, including antibody titers against herpes simplex virus, Epstein-Barr virus, cytomegalovirus (CMV), respiratory syncytial virus, human immunodeficiency virus, Mycoplasma pneumoniae, Campylobacter jejuni/coli and cardiolipin. These studies showed the presence of antibodies indicative of a CMV primary infection in 22% cases and of a Campylobacter jejuni/coli infection in 13%. Co-infection was observed in 3 cases. Serology remained negative in 12 patients with a preceding respiratory infection. There was no correlation between serology and the severity of the disease. Absence of antecedent events and of positive anti-infectious serology was observed in only 10 patients.

No evidence of auditory dysfunction in guinea pigs immunized with myelin P0 protein.

Recent data have focused on the peripheral nerve myelin glycoprotein P0 as a putative autoantigen involved in the autoimmune etiology of some cases of Meniere's disease, idiopathic sensorineural hearing loss and sudden deafness. To determine whether antibodies to myelin P0 can alter cochlear function, 13 healthy guinea pigs were immunized with purified porcine myelin P0 while 10 controls were injected with saline water. The animals were then evaluated for evidence of evolving inner ear disease using immunological, electrophysiological and morphological methods. Twenty-six experimental ears were tested weekly with a brainstem auditory evoked potential technique for a period of 4 months and were compared to 20 control ears. Uniformly, all P0-sensitized guinea pigs showed antibodies to myelin protein P0 as evidenced by ELISA. Clinical signs of inflammatory demyelination were not discernible in P0-sensitized guinea pigs and all the animals were qualitatively normal. No significant increase of evoked potential thresholds was found in the P0-sensitized animals when compared to controls (P>0.05). Peak latencies of waves I, II, III, IV and V and inter-peak latencies in P0-sensitized guinea pigs did not significantly differ from those of controls (P>0.05). Histological sections of inner ear and peripheral nerves were free of disease in both groups. These findings indicate that the sole presence of antibodies to myelin P0 in the sera of guinea pigs or patients suspected of having autoimmune inner ear diseases is unlikely to elicit auditory abnormalities and that additional factors are necessary for the pathogenic development of these disorders.

Autoimmune sensorineural hearing loss in young patients: an exploratory study.

OBJECTIVES: P0 protein is expressed exclusively in myelinating Schwann cells of the peripheral nervous system. In a previous study from our laboratory, 27% of patients with sensorineural hearing loss (SNHL) had antibodies to P0 protein in their serum. The purpose of the present exploratory study was to examine the relationship between the clinical presentation of SNHL among children and young adults (age range, 5-30 y) and the presence of serum anti-P0 antibodies. STUDY DESIGN: The data were collected by retrospective questionnaires from Belgian otolaryngologists. METHODS: Patients were divided for comparison into two groups according to the presence or absence of anti-P0 antibodies. RESULTS: Analyses of clinical data and audiometric results indicated that a progressive hearing loss was more frequently recorded in the patients in the anti-P0 antibody-positive group (82% [14 of 17]) than in those in the anti-P0 antibody-negative group (35% [6 of 17]) (P <.005). CONCLUSIONS: Thus, in the age group in the present study, autoimmune SNHL (as measured in the present study by the presence of anti-P0 antibodies) is more frequently associated with progressive than with sudden hearing loss. The implications of this finding for preventive screening of hearing loss in children and young adults are discussed.

Guinea pig inner ear antigens: extraction and application to the study of human autoimmune inner ear disease.

In this study, the authors attempted to develop a method of extracting guinea pig inner ear antigens for otoimmunological research, and to investigate the distribution of the antigens in the various structures of the inner ear. The antigens were extracted either from the entire or from various parts of the guinea pig inner ear. These antigens were separated on sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. Western blot techniques were used to test sera from patients with inner ear disease against guinea pig inner ear protein extracts. It found that the various molecular weight antigens in the inner ear were associated with the different structures of the inner ear. The sera of 37.5% (N = 80) of patients reacted with two bands (30 and 58 kd) of the guinea pig inner ear immunoblots. The 58 kd band was not specific to the inner ear, but instead was also found in the immunoblots of other guinea pig tissues (brain, lung, and liver). This study suggests that the various antigens of interest could be further extracted and purified from the corresponding locations of the inner ear.

Clinical significance of antiproteinase 3 antibody positivity in cANCA-positive patients.

We addressed the clinical significance of antiproteinase 3 (anti-PR3) antibody (Ab) positivity by reviewing the files of 79 patients whose serum contained antineutrophil cytoplasmic antibodies with a cytoplasmic staining pattern (cANCA) and had been tested for anti-PR3 reactivity. Vasculitis was present in most (22/35) cANCA+ PR3+ patients but in only a few (5/44) cANCA+ PR3- patients, thereby suggesting that anti-PR3 Ab positivity in cANCA+ patients is more indicative of vasculitis than cANCA positivity alone. Noteworthy, one-third of cANCA+ PR3+ patients -- those with anti-PR3 Ab titres lower than 100 U/ml -- did not suffer from vasculitis. Anti-PR3 reactivity in vasculitis patients was only weakly associated with Wegener's granulomatosis (WG), as nine out of 22 cANCA+ PR3+ vasculitis patients (41%) did not fulfil the ACR classification criteria for WG. There was no correlation between anti-PR3 Ab titres and disease activity at diagnosis. However, titres measured when patients were in remission were much lower than initial values. Taken together, our results indicate that anti-PR3 Ab positivity should be interpreted in its clinical context.

COCH5B2 is a target antigen of anti-inner ear antibodies in autoimmune inner ear diseases.

OBJECTIVE: This study was designed to identify the 58-kDa inner ear protein against which the sera of some patients with idiopathic, progressive sensorineural hearing loss or Ménière's disease strongly react. BACKGROUND: We and other groups have previously demonstrated that a 58-kDa protein extracted from guinea pig or bovine inner ear tissue is a target of antibodies in serum samples from some patients with autoimmune inner ear diseases. METHODS: After separation of inner ear proteins by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, the bands corresponding to 58 kDa were localized and excised from the gel. The concentrated protein was then digested with trypsin, and the peptide fragments were separated by high-pressure liquid chromatography. Three fractions were subjected to amino acid sequencing by the classic Edman degradation. RESULTS: The sequence of a stretch of 14 amino acids of the first fragment was identical to that of amino acids 526 to 539 of the COCH5B2 protein. The sequences of 11 and 10 amino acids of the second and third fragments, respectively, also were identical to residues 417 to 427 and 396 to 405 of the COCH5B2 protein. These data, together with two-dimensional gel electrophoresis followed by Western blot experiments, confirmed that the 58-kDa inner ear protein is the COCH5B2 protein. DISCUSSION: These findings indicate that the 58-kDa target protein of antibodies in serum samples of patients with autoimmune inner ear diseases is the COCH5B2 protein, a molecule that is highly and specifically expressed in the cochlea and vestibule.

Modelling of the 3-D structure of IFN-alpha-k and characterization of its surface molecular properties.

The 3-D structure of IFN-alpha-k (one of the alpha-interferon family) was constructed and optimized by molecular modelling starting from the X-ray structure of IFN-beta. The molecular surface of the optimized 3-D structure of IFN-alpha-k was then evaluated and characterized for its hydrophobicity/hydrophilicity. The structure of IFN-alpha-k was completed with its first segment (23 amino acid residues) called signal peptide. The 3-D structure of this segment was predicted to be in helical form bonded to the core by one loop. It was found that the complete structure of IFN-alpha-k can exist in at least two main conformations as far as the orientation of the signal peptide is concerned, i.e. in the open form (in which the signal peptide is directed outward of the 'body' of the molecule) and the closed form (where the signal peptide is aligned with the body). The relative stability of these forms strongly depends on the stabilization by the environment (e.g. by solvation) due to the prevailing hydrophilicity of the body and hydrophobic character of the signal peptide.

HLA class II-associated genetic susceptibility in idiopathic progressive sensorineural hearing loss.

To investigate the association between genes in the major histocompatibility complex and inner ear disease susceptibility at the DNA level, high-resolution genotyping for HLA class II (HLA-DR, -DQ, -DP) was performed by polymerase chain reaction-sequence-specific oligonucleotide reverse dot blot and polymerase chain reaction-restriction fragment length polymorphism analysis in 34 patients with idiopathic progressive sensorineural hearing loss (PSHL) and in 214 controls. The frequencies of DRB1*0301, DRB3*0101, DQB1*0201, and DPB1*0401 were significantly increased in patients with idiopathic PSHL compared with controls. The DQB1*0301 allele was significantly decreased in the patients. A linkage disequilibrium was probably responsible for the concomitant increase of both DRB1*0301 and DRB3*0101 alleles in patients. The increase of DQB1*0201 in patients was associated with the DRB1*0301 allele. In addition, the telomeric DPB1*0401 allele may act as an independent risk factor. The DQB1*0301 allele may have a protective role in the pathogenesis of idiopathic PSHL. These results suggest that the specific HLA class II gene products may confer susceptibility or resistance to idiopathic PSHL.

Inner ear autoantibodies and their targets in patients with autoimmune inner ear diseases.

Immunological mechanisms are thought to play an important role in the pathogenesis of some cochleo-vestibular diseases. This study attempts to present further evidence of autoantibodies reactive against guinea pig inner ear proteins found in patients with autoimmune inner ear diseases (AIED) and specifically identifies the main target antigens of these antibodies. Sera from 110 patients with a clinical diagnosis of either rapidly progressive sensorineural hearing loss (n = 32). Ménière's disease (n = 41), sudden deafness (n = 6) or other aetiologies of hearing loss (n = 11) were screened by the Western blot technique. Forty-four percent of the patients' sera had antibodies to several inner ear proteins, of which the 30, 42 and 68 kDa proteins were found to be the most reactive. These highly reactive proteins were identified by gas-phase micro sequencing after digestion with trypsin and separation of peptide fragments by high-performance liquid chromatography. A partial sequence of each protein was determined. These data, together with those obtained from 2-dimensional gel electrophoresis followed by Western blotting, demonstrated that the 30 and 42 kDa inner ear proteins are the major peripheral myelin protein P0 and the beta-actin protein, respectively, while sequence analysis indicated that the 68 kDa protein is novel. These findings further support the hypothesis that several populations of antibodies may contribute to the enhanced immunological activity of AIED patients. They also add a new dimension to our knowledge of AIED and may open new avenues in the development of simple serological assays, which are easier to perform and more rapid than Western blotting.