Pepsinogen identification in the middle ear fluid of children with otitis media with effusion.

OBJECTIVES: This study aims to investigate the presence and concentration of pepsin/pepsinogen in middle ear fluid and to discuss the potential mechanisms involved in the pathogenesis of this condition. PATIENTS AND METHODS: A total of 33 children (21 boys, 12 girls; mean age 5.7±2.4 years; range 3 to 13 years) diagnosed with otitis media with effusion and scheduled for operation were enrolled into the study. Fluids aspirated from the middle ear were assessed for the presence of pepsinogen and albumin and blood samples were drawn simultaneously for comparison. RESULTS: Mean pepsinogen concentration was statistically significantly higher in middle ear fluids compared with serum samples (262.4 ng/mL [range: 211.7 ng/mL - 301.1 ng/mL] versus 102.6 ng/mL [range: 80.7 ng/mL - 134.5 ng/mL], respectively) (p<0.001). On the other hand, mean albumin concentration was significantly lower (1.1 g/dL [range: 0.01 g/dL - 9.5 g/dL] versus 5.8 g/dL [range: 0.9 - 9.5 g/dL], respectively) (p<0.001). The highest pepsinogen concentration was detected in patients with purulent effusion (275.3 ng/mL). CONCLUSION: Our findings support the theory of gastro-esophageal reflux related pepsinogen transition to the middle ear and indicate that pepsinogen may a reliable biochemical marker for the assessment of gastro-esophageal reflux.

[Microbial view and optimal antibiotic therapy in acute purulent ENT diseases]. / Mikrobnyi peizazh i puti ratsional'noi antibiotikoterapii pri ostrykh gnoinykh zabolevaniiakh LOR-organov.

The aim of the study was elaboration of optimal antibiotic treatment in acute pyoinflammatory ENT pathology basing on the results of updated microbial tests and basic rules of microflora suppression by antibiotics. A total of 200 patients were examined. Of them, acute purulent sinusitis was diagnosed in 15, acute purulent otitis media in 25, paratonsillar abscess in 12 cases. Their microflora was studied for antibiotic sensitivity, the ability to form beta-lactamase was evaluated, microbiological examination of the content from purulent foci was made. It was found that the majority of acute purulent ENT diseases were caused by streptococci, gram-negative microorganisms (H. influenzae, M. catarrhalis, Klebsiella spp. et al), staphylococci and obligate anaerobes most of which produce beta-lactamase (up to 79%). From 88 to 100% pathogens were sensitive to amoxicillin/clavulanate, 18-59% -- to cephasoline, lincomycin or ampicillin. The treatment with antibiotics of different groups was assessed by dynamics of clinical symptoms and results of repeated microbiological analyses. The optimal antibiotic to start therapy of patients with acute purulent ENT diseases was amoxicillin/clavulanate in a new form -- 1000 mg (twice a day for 7 days).

Alpha 1-antitrypsin and ilomastat inhibit inflammatory proteases present in human middle ear effusions.

OBJECTIVES: Proteases of both the serine and metalloproteinase families have been shown to play a role in the pathogenesis of otitis media (OM). Inhibitors of proteases from each of these families have been shown to beneficially impact disease progression in a number of related chronic inflammatory conditions, but their use has not been studied in OM. The purpose of this study was to assess the activity of the protease inhibitors recombinant alpha 1-antitrypsin (rAAT) and ilomastat on inflammatory proteases present in human middle ear effusions (MEEs), with a view to their potential utility in the treatment of OM. STUDY DESIGN: Prospective and ex vivo. METHODS: MEEs were collected from 100 patients presenting for middle ear surgery, most commonly tympanostomy tube placement or treatment of acute posttympanostomy otorrhea (APTO). MEEs were analyzed for the presence of matrix metalloproteinases (MMPs) and human neutrophil elastase (HNE) and the inhibitory activity of rAAT and ilomastat on these proteases, respectively. RESULTS: MMP levels were highest in APTO, and HNE was highest in chronic suppurative OM and APTO. High levels of MMP and HNE (>3 mAU/min) were found in 52% and 37% of MEEs, respectively. Ilomastat and rAAT demonstrated significant inhibition of MMP and HNE activity (>30% reduction), respectively, in 80% and 82% of MEEs with high levels of activity. CONCLUSIONS: Proteases are commonly found in OM. Ilomastat and rAAT are potent inhibitors of proteases in MEEs across a wide range of OM in humans. Investigation into the potential therapeutic benefits of these protease inhibitors is warranted.

[Oxidative profile of tonsil infection. Study of antioxidant enzymes in tonsil and blood]. / Perfil oxidativo de la infección amigdalar. Estudio de las enzimas antioxidantes en amígdala y sangre.

In order to investigate the effect of oxidative damage due to free radicals on ENT infectious diseases, levels of superoxide dismutase (SOD), glutathione-peroxidase (GPx) and reductase (GRt) and the total antioxidant status (TAS) were measured by spectrophotometry on tonsillar tissue obtained from tonsillectomy in 538 patients, who were divided in three groups according to their surgical indication: tonsillar hypertrophy (n = 235), recurrent tonsillitis (n = 280) or peritonsillar abscess (n = 23). SOD concentration were also measured on adenoid tissue and middle ear exudate in 75 patients from the first two groups. Erythrocyte and tonsillar SOD levels were significantly greater in the abscess group, and lower in the hypertrophic one. These differences were similar for GPx and TAS. For GRt, its level in abscess were lower than in the other two groups in a statistically significant way. There were strong correlations between erythrocyte and tonsillar SOD, tonsillar SOD and GPx, tonsillar SOD and TAS, and tonsillar GPx and TAS. SOD concentrations from adenoid tissue and middle ear exudate did not affect its blood level. So, we can conclude that tonsillar oxidative damage is determined by the frequency or the severity of local infections, and it can be evaluated by measuring the SOD concentration in the tonsillar tissue or in the peripheral blood. So, it can be considered a good marker of tonsillar damage.

Neutrophil elastase and its complex with alpha 1-antitrypsin in the pathogenesis of chronic suppurative otitis media.

Neutrophil elastase (NE) and its complex with alpha 1-antitrypsin were quantified in ear discharges from 15 patients with chronic suppurative otitis media (CSOM), and their levels were compared to those in middle ear effusions from 10 pediatric patients with chronic otitis media with effusion (OME). The localization of immunoreactive NE was also examined by immunocytologic study. The mean value of total NE was 161.8 +/- 29.5 micrograms/mL in CSOM, which was significantly higher than that in OME (16.5 +/- 6.7 micrograms/mL). The mean value of NE-alpha 1-antitrypsin complex was 13.7 +/- 8.8 micrograms/mL in CSOM and 7.8 +/- 4.0 micrograms/mL in OME. There was no significant difference between culture-positive and culture-negative samples in CSOM. Immunoreactive NE could be observed in the neutrophils of ear discharges, and extracellular release of NE was also observed. Ear discharges in CSOM contain a considerable amount of uncomplexed NE, of which the level does not depend on active bacterial infections. Uncomplexed NE seems to play a role in the chronicity of CSOM.

Quantitative bacterial cultures and beta-lactamase activity in chronic suppurative otitis media.

Aspiration of the exudate through open perforation was performed in 54 children with chronic suppurative otitis media. Eighty aerobic and 81 anaerobic isolates were recovered. Aerobic bacteria only were involved in 20 patients (37%), and anaerobic organisms only in seven (13%). Mixed aerobic and anaerobic isolates were recovered from 27 patients (50%). The most common bacteria isolated were anaerobic gram-positive cocci, Bacteroides melaninogenicus group, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Staphylococcus aureus. There were 45 beta-lactamase-producing bacteria (BLPB), 30 aerobes, and 15 anaerobes recovered from 38 patients (70%). beta-Lactamase activity was detected in 30 of the 38 ear aspirates (79%) that contained BLPB. All but one of these organisms were in excess of 10(4) colony-forming units/mL. The detection of beta-lactamase activity in the ear aspirates provides evidence of the role of BLPB in the failure of penicillin therapy to eradicate chronic ear infection.

Hydrolase activity in acute otitis media with effusion.

Biochemical studies of middle ear effusions (MEE) from patients with chronic or recurrent otitis media with effusion (OME) have demonstrated the presence of significant levels of certain hydrolytic and oxidative enzymes. We have examined MEE from patients with acute OME for the content of a number of lysosomal hydrolases and find no significant differences in the mean values for acid phosphatase, alpha-mannosidase, beta-galactosidase, beta-glucuronidase, hexosaminidase, and neuraminidase between purulent and serous effusions. In every case, the mean activities of these enzymes were greater in culture-positive than in culture-negative effusions although this difference was significant only in the case of neuraminidase. Neuraminidase activity was detected in 78% of those MEEs from which Streptococcus pneumoniae could be cultured and in only 32% to 64% of all other effusions. No correlation was observed between the level of neuraminidase released into the extracellular growth medium and the infectivity of various strains of S pneumoniae.

Biochemical pathology of otitis media with effusion.

The sequential cytologic and biochemical events of middle ear effusion (MEE) were studied in experimental models of serous otitis media (SOM) and purulent otitis media (POM) in chinchilla. In the SOM model, the initial appearance of neutrophils was followed by macrophages. In the POM model, neutrophils were the predominant cells in MEE and the number of neutrophils was about 100-fold higher than in the SOM model. The activity of lysozyme in MEE was higher in POM than in SOM and correlated with the number of neutrophils and mononuclear phagocytes. The results of the present study suggest that neutrophils and mononuclear phagocytes are one of the main sources for lysozyme levels in MEE during otitis media.

The ceruminous glands in chronic suppurative otitis media. A histological and histochemical study.

The ceruminous glands have been studied histologically and histochemically in twenty patients suffering from chronic suppurative otitis media with discharging ears, and in ten patients with dry central perforations, in comparison with normal ears. The present study revealed hyperactivity of the glands initially; later on, atrophy and fibrosis took place. Thus the diminution of wax may be due to: (1) The washing effect of the discharge. (2) Atrophy of the glandular structures. (3) Periglandular fibrosis. As the discharge stops, regeneration of the glands occurs.

Lysozyme activity and immunoglobulins in middle ear effusion fluid in acute purulent otitis media and in otitis media with effusion.

147 samples of punctured middle ear effusion fluid from cases of otitis media with effusion and 150 samples from patients with acute purulent otitis media were tested for lysozyme activity. In otitis media with effusion the concentration was 182.0 U/ml, in acute otitis 433.8 U/ml. The lysozyme concentration in otitis media with effusion depended upon the nature of the effusion. Serous fluid showed an activity of 124.8 U/ml and mucoid 311.6 U/ml, respectively. In culture-positive cases of acute otitis media the lysozyme level was 423.4 U/ml. Culture-negative cases showed about the same concentration, 438.3 U/ml. The possible role of lysozyme in defence systems of the middle ear is discussed.