Enhanced Expression of Plasminogen Activators and Inhibitor in the Healing of Tympanic Membrane Perforation in Rats.

The significance of plasminogen activation during the tympanic membrane (TM) healing is known mainly from studies performed on knock-out mice. In the previous study, we reported activation of genes coding proteins of plasminogen activation and inhibition system in rat's TM perforation healing. The aim of the present study was the evaluation of protein products expressed by these genes and their tissue distribution using Western blotting and immunofluorescent method, respectively, during 10-day observation period after injury. Otomicroscopical and histological evaluation were employed to assess the healing process. The expression of urokinase plasminogen activator (uPA) and its receptor (uPAR) were significantly upregulated in the proliferation phase, with subsequent gradual attenuation during remodeling phase of healing process, when keratinocyte migration was weakening. The expression of plasminogen activator inhibitor type 1 (PAI-1) also showed the highest levels during the proliferation phase. The increase of tissue plasminogen activator (tPA) expression was observed during the whole observation period, with the highest activity during the remodeling phase. Immunofluorescence of these proteins was present mainly in migrating epithelium. Our study found that plasminogen activation (uPA, uPAR, tPA) and inhibitory (PAI-1) molecules form a well-structured regulatory system of the epithelial migration that is critical to the healing of TM after its perforation.

The effects of adenoidectomy of serum insulin-like growth factor-1 (IGF-1) and ghrelin in hypertrophied adenoids in children with otitis media with effusion.

<b>Aim:</b> The aim of the current study was to assess the serum levels of insulin-like growth factor-1 (IGF-1) and ghrelin in hypertrophied adenoids in children suffering with or without otitis media with effusion before and after adenoidectomy. <br><b>Material and methods:</b> Serum IGF-1 and ghrelin concentrations were measured with specific enzyme-linked immunoassay (ELISA) methods. The study was carried out in 20 children with otitis media with effusion. The reference group comprised 24 children with hypertrophied adenoid, while control group included 19 children. <br><b>Results:</b> This mean values of IGF-1 in children with otitis media with effusion and children with hypertrophied adenoid before adenoidectomy were significantly lower than those found in healthy children. Serum levels of IGF-1 were higher after adenoidectomy. There was a significant difference of serum ghrelin levels between both examined groups and the control group. <br><b>Conclusion:</b> Our results suggest that adenoidectomy in children with hypertrophied adenoids and in children with otitis media with effusion significantly increases the level of IGF-1 in serum compared to before surgery through the effect of the GH-IGF-1 axis, which could contribute to children's growth.

Vascular endothelial growth factor and transforming growth factor ß in hypertrophic adenoids in children suffering from otitis media with effusion.

OBJECTIVES: The study objective was to assess the levels of VEGF-A and TGF-ß cytokines in the children with adenoid hypertrophy concomitant with exudative otitis media (OME) and in children with adenoid hypertrophy (HA) alone. METHODS: The study material consisted of hypertrophic adenoids removed during adenoidectomy from 39 children (20 girls and 19 boys), aged 2-7 years suffering from OME. The reference group included 41 children (19 girls and 22 boys), aged from 3 to 9 years with adenoid hypertrophy. The levels of VEGF-A and TGF-ß were determined in supernatants obtained from phytohemagglutinin-stimulated cell cultures of the adenoids using a commercial enzyme-linked immunosorbent assay kit. RESULTS: The median VEGF-A and mean TGF-ß concentrations in the study group were significantly higher than those in the reference group (503 pg/mL versus 201 pg/mL, P < 0.001 and 224 pg/mL versus 132 pg/mL, P < 0.001, respectively). ROC analysis revealed that the area under the curve (AUC) for VEGF-A was 0.952 with diagnostic sensitivity and specificity of 95%, whereas for TGF-ß it was 0.902 with 60% sensitivity and the same specificity as for VEGF-A. There was no significant difference between the AUC for VEGF-A and TGF-ß (P = 0.573). CONCLUSIONS: The changes in the levels of VEGF-A and TGF-ß may indicate bacterial pathogen as one of the causes of exudative otitis media in children. Determination of VEGF-A and TGF-ß could be used as additional and objective tests to confirm the clinical diagnosis.

Two unrelated families with variable expression of Fraser syndrome due to the same pathogenic variant in the FRAS1 gene.

We report on two unrelated families of Polish origin with variable expression of Fraser syndrome (FS; MIM#219000) due to homozygosity for the same pathogenic variant, c.6963_6964dup, of FRAS1. In one family, the disorder presented with perinatal and prenatal lethality. One affected female from family 2 who was followed-up for 32 years, represented a relatively favorable long-term outcome. She displayed the typical craniofacial dysmorphism, including right cryptophthalmos, cutaneous syndactyly, abnormalities of the stomathognatic system, bilateral atresia of the external ear canals resulting in conductive hearing loss, and malformations of the larynx, spleen, kidney, and genitourinary tract. Her intellectual capacities were normal. Our observations illustrate that expression and severity of FS, even when caused by the same pathogenic variant, may be quite different ranging from a lethal disorder to a condition with multiple physical malformations but normal psychomotor development. In addition, we propose that the FRAS1 c.6963_6964dup variant may be a founder mutation in the Polish population. Therefore, it would be reasonable to test specifically for this variant first in any FS1 patient of Polish ancestry.

Selected cytokines in hypertrophic adenoids in children suffering from otitis media with effusion.

OBJECTIVES: The aim of the current study was to assess the levels of MMP-8, MMP-9 and TIMP-1 in the group of children with adenoids who suffered from exudative otitis media. METHODS: The study included 20 patients (10 females and 10 males) with adenoid hypertrophy coexisting with otitis media with effusion. The reference group included 24 patients (10 females and 14 males) with adenoid hypertrophy without otitis media. The levels of MMP-8, MMP-9 and TIMP-1 were determined in supernatants obtained from phytohemagglutinin (PHA)-stimulated cell cultures of the tonsils, using commercial enzyme-linked immunosorbent assay kits (R@D Systems, USA). RESULTS: The median MMP-8, MMP-9 and TIMP-1 concentrations (220.8 ng/mL, 311.1 ng/mL, 53.5 ng/mL, respectively) in the study group were significantly higher (p = 0.000, p = 0.000, p = 0.048, respectively) than those in the reference group (93.5 ng/mL, 112.5 ng/mL, 36.95 ng/mL, respectively). ROC analysis revealed that the area under a curve (AUC) for both metalloproteinases MMP-8 and MMP-9 was 1 with a diagnostic sensitivity of 100% and diagnostic specificity of 95.8%, as compared to 0.690 for TIMP-1. Significant differences were found between the AUC for MMP-8 and TIMP-1 and MMP-9 and TIMP-1 (p < 0.001 for both comparisons). CONCLUSIONS: The changes in the concentrations of MMP-8, MMP-9 and TIMP-1 may indicate an increased remodeling of the extracellular matrix in children with adenoid hypertrophy and otitis media with effusion. The findings can have clinical as well as diagnostic utility. Determination of MMP-8 and MMP-9 may help qualify a child for adenoidectomy and differentiate pediatric patients affected by adenoid hypertrophy with and without otitis media.

Expression of collagens type I and V in healing rat's tympanic membrane.

OBJECTIVES: Our preliminary study performed on perforated rat's tympanic membrane (TM) using Rat Wound Healing RT2 Profiler PCR Array showed significantly increased levels of mRNA for collagens type I and V. Enhanced expression of those genes does not assure that their protein products are indeed present, and in what quantity. Therefore, this study was undertaken to analyze the collagen type I and V content in the healing TM. METHODS: Sixty rats were used, of which 10 served as controls and the others had their TM perforated. The experimental animals were divided into five subgroups on the basis of time points (03, 06, 09, 14, 20 day after injury). Videootoscopy and histology were employed to assess the morphology of the healing process. The expression of collagen type I and V was evaluated using Western blot analysis. Tissue localization of collagens was determined by the immunofluorescence method. RESULTS: The collagen type I expression was three times higher on the third day after injury and remained on that level for whole period of observation, up to day 20. The increase of the collagen type V expression was gradual, reaching the highest level on day 14 following injury. In comparison to the control TM statistically significant increase in the level of expression was observed starting from day 09 to the end of observation period. In healing TM immunofluorescent labeling of collagen type I and V was seen on the surface of remnants of previous lamina propria and in the loose proliferating fibrous tissue. On day 20 immunofluorescence was present mainly on the surface of thin connective tissue layers forming the scar in the place of previous perforation. CONCLUSION: Although the collagens type I and V are present only in subepithelial layer in the normal rat's TM they play significant role in TM healing process.

A 23-year follow-up of a male with Hajdu-Cheney syndrome due to NOTCH2 mutation.

We present a natural history of a 32-year-old man with Hajdu-Cheney syndrome (HJCYS), because of the de novo truncating mutation in the exon 34 of NOTCH2 (c.6424-6427delTCTG, p.Ser2142ArgfsX4), who has been followed up for a period of 23 years (between 9 and 32 years). During follow-up, we observed abnormalities of vision, hearing, voice, and progression of craniofacial features in the form of skeletal dysplasia with affected skull, dentition, spine, limbs, fingers, and toes. Low bone mineral density and history of fragility fractures also suggested primary osteoporosis being a clinical manifestation. According to Stengel-Rutkowski, Schimanek, and Wernheimer (1984; Human Genetics, 6, 272-295), systematic data acquisition has been used for quantitative analysis of anthropological, radiographic, and clinical features at childhood, adolescence, and young adulthood separately. A detailed phenotype description together with the results of reanalysis of 14 reports so far published on patients with HJCYS and NOTCH2 mutation showed similar phenotype evolution with age. The spectrum of observed features may improve diagnostic tools for HJCYS at different periods of the lifespan.

Presentation, diagnosis and management of neck abscesses in children.

UNLABELLED: The aim of the study was to analyze the presenting signs and symptoms, diagnostic procedures, clinical course, pathogenic organisms and management of neck abscesses in children. MATERIALS AND METHODS: This retrospective medical records review comprised of 51 pediatric cases referred because of neck abscess from 2001 to 2014. Medical records of the patients were reviewed for demographic data, clinical presentation, treatment before referral, localization of the abscess, imaging evaluation, medical and surgical treatment, bacteriological data and complications. RESULTS: Average age of the patients was 4.9 years. 18 (35%) of the children were below one year of age. The submandibular was the most common area involved (41,2%). Contrast-enhanced computed tomography (CECT) was performed in 45% of patients mainly with deep neck abscesses. All CECT scans showed the fluid collections. In all but one of the patients treated surgically and diagnosed with ultrasound and/or CECT surgical intervention revealed presence of pus. The most common pathogen was Staphylococcus aureus- SA (78% of all isolates) of which 24% were methicillin resistant Staphylococcus aureus - MRSA. Clindamycin resistance was detected in 28% of all SA isolates and in 67% (4/6) of all MRSA isolates. All MRSA isolates were sensitive to vancomycin. CONCLUSIONS: CECT is reliable imaging technique for diagnosing deep neck abscesses in children. Infants with neck abscesses are at higher risk of having MRSA as offending pathogen which should be taken into consideration when considering empiric treatment. Vancomycin is recommended as empiric antibiotic therapy in newborns with neck abscess.

Enhanced expression of hepatocyte growth factor in the healing of experimental acute tympanic membrane perforation.

OBJECTIVES: The present study was performed to investigate the expression of hepatocyte (HGF), epidermal (EGF) and vascular endothelial (VEGF) growth factors in the course of healing of experimental tympanic membrane (TM) perforations in rats. The goal was to explain the role of these growth factors in the healing process of TM and to assess the possibility of their future application as healing promoters. METHODS: Seventy rats were used, of which 10 served as controls and the others had their TM perforated. The experimental animals were divided into six subgroups on the basis of time points (01, 03, 05, 07, 09, 15 day after injury). Videootoscopy and histology were employed to assess the morphology of the healing process. The expressions of HGF, EGF and VEGF were evaluated using Western blot analysis. Tissue localization of HGF was determined by the immunofluorescence method. RESULTS: HGF was hardly detectable in normal TM; however, a significant increase was noted in its expression starting from the third day after injury throughout the follow-up period, with the highest level on day 05. The analysis of HGF tissue localization with immunofluorescence revealed diffuse staining in the cytoplasm of proliferating epithelial cells. The expression of EGF was elevated on the first day after injury, not reaching statistical significance, and then returned to the level observed in the control TM. No significant differences were noted in the expression of VEGF. CONCLUSION: High expression of HGF during the healing process of acute TM perforations makes it a promising candidate for further studies oriented towards its possible use in augmentation of TM healing.

[The healing process of tympanic membrane perforations in rats]. / Proces gojenia perforacji blon bebenkowych u szczurów.

INTRODUCTION: Tympanic membrane (TM) perforations are commonly seen in clinical practice as a result of trauma or in the course of otitis media. The TM is a unique structure suspended in air which makes its healing processes different than in the skin wounds. The aim of the study was otoscopical and histological evaluation of the rat's TM healing process. MATERIAL AND METHODS: 56 male Wistar rats were used for the study. Fifty of them had TMs perforated bilaterally using CO2 laser, additional 6 served as a controls. The animals were sacrificed on either day 1, 2, 3, 6 and 10 post injury. Process of healing was assessed otoscopicaly, subsequently TM were dissected and processed for histological evaluation. RESULTS: At day 6 after perforation half and on day 10 all of TM were healed. On the first day, in histological evaluation, focal thickening of the epithelial layer was observed at some distance from the edge of perforation, on the side of annulus. On the following day proliferation of epithelium covering outer surface of TM on the side of the malleus handle and annulus was clearly visible. An eosinophilic mass containing macrophages and granulocytes was seen in front of the migrating epithelium. On day 3-6 migrating epithelium reached the edge of perforation. Proliferation of the connective tissue layer followed the epithelium. CONCLUSIONS: The present results indicate that the squamous epithelium covering the outer surface of TM constitutes the first layer which restores continuity of TM. The proliferation of the connective tissue occurs in the direct vicinity of the proliferating and migrating epithelium.

Expression of Toll-like receptors on peripheral blood white cells in acute otitis media.

OBJECTIVE: From 10 to 15% of children suffer from recurrent acute otitis media (AOM). An association between polymorphism in TLRs and their co-receptor CD14 with otitis media proneness has been described in children. Moreover, the experiments on animal models have shown that TLRs and their signaling molecules are critical for timely resolution of bacterial otitis. AIM: The aim of this study was to determine the expression of TLR1, TLR2 and TLR4 on lymphocytes, monocytes and granulocytes in peripheral blood in children with recurrent or persistent AOM. METHODS: The study was performed on a group of 25 children hospitalized for recurrent AOM, failures of previous treatments and/or acute mastoiditis. The results were compared to the control group of healthy children at the same age. The expression of TLRs on peripheral blood white cells was measured by flow cytometric analysis. The results were expressed as mean fluorescence intensity (MFI). The statistical analysis was performed using the Mann-Whitney U test. RESULTS: The highest expression of TLR was found on monocytes, the lowest on lymphocytes in both groups of children (AOM and the control one). The expression of TLR1 was the lowest and expression of TLR4 was the highest on all examined cells. The expression of all examined TLRs on monocytes was significantly higher in the AOM group. CONCLUSIONS: Peripheral blood monocytes are characterized by increased expression of TLRs in the course of recurrent AOM.

Production of cytokines by mononuclear cells of hypertrophic adenoids in children with otitis media with effusion.

Hypertrophic adenoids with otitis media with effusion is a common infectious disease and present a serious otological problem in children. Cytokines, potent inflammatory mediators, play important role in the initiation of immunological response in otitis media. Adenoids excised due to hypertrophy with or without chronic otitis media with effusion were used to isolate mononuclear cells. Secretion of cytokines by non-stimulated and PHA-stimulated cells was determined by specific ELISAs. We found a significant increase in the production of IL-5 and TNF-α secreted by adenoidal cells of children with otitis media with effusion compared to group with hypertrophic adenoids. No differences were found in the secretion of IL-8, IL-6, and IL-10 between these two groups of patients. Our results suggest a difference between the immunological responses in the course of hypertrophic adenoids with otitis media as compared to hypertrophic adenoids.

Increased percentage of T cells with the expression of CD127 and CD132 in hypertrophic adenoid in children with otitis media with effusion.

The hypertrophic adenoid may promote chronic suppurative otitis media in children as it fulfills its immune function. The number of lymphocytes in the adenoid and their cooperation in the immune response depend of on their proliferation and migration to the effector sites. Interleukin 7 (IL-7) is essential for the normal development and function lymphocytes. IL-7 plays pivotal role for activation and proliferation of T and B cells. The heterodimeric interleukin-7 receptor (IL-7R) is composed of the IL-7Rα (127) and the common cytokine receptor γc (CD132). The aim of this study was to evaluate the percentage of lymphocytes T (CD4(+) and CD8(+)) with IL-7R (CD127 and CD132) expression in hypertrophic adenoid in children suffering with otitis media with effusion for a duration of 3 months. Adenoid excised due to hypertrophy with or without chronic otitis media with effusion was used as study material. CD4(+) CD127(+), CD4(+)132(+), CD8(+)CD127(+) and CD8(+)CD132(+) cell subpopulations were identified using monoclonal antibodies and flow cytometry. The percentage of CD4(+) and CD8(+) T cells with CD127 receptor expression in hypertrophic adenoid of children with otitis media with effusion was statistically significantly higher than in hypertrophic adenoid group. The percentage of CD4(+) T cells with CD132 expression in the study group was statistically significantly higher than in the reference group. The percentage of CD8(+) T cells with CD132(+) expression was not statistically different in both groups. The increased percentage of T lymphocytes with IL-7R expression (CD127 and CD132) in hypertrophic adenoid seems to influence the quantity of lymphocytes and upset the immunological function of tonsils which can influence the course of otitis media with effusion.

[Influence of tympanic membrane changes on immittance and extended frequency audiometric findings]. / Wplyw zmian strukturalnych blony bebenkowej na wyniki audiometrii impedancyjnej i tonalnej w rozszerzonym zakresie czestotliwosci.

INTRODUCTION: The aim of the study is to analyze the relationship between otoscopic tympanic membrane abnormalities, results of impedanace and extended-high-frequency audiometry in subjects with history of treatment of secretory otitis media (SOM) and ventilation tube insertion. MATERIAL AND METHODS: 97 subjects treated because of SOM were examined in years 1999-2000. Videootoscopic examinations, extended-high-frequency audiometry and tympanometry with ispilateral acoustic reflex were performed in each patient. The results were analysed in three groups: no otoscopic abnormalities (BZO), retraction pockets (KR) and atrophy and myringosclerosis (AM). The results were compared to otologicaly healthy control group in the same age. RESULTS: The most common tympanic membrane abnormality were focal atrophy (64.7%) of ears and myringosclerosis (37.2%). Mean pure-tone audiometric threshold were significantly higher in groups KR and AM than in control and BZO groups. Low degree of positive correlation was found between the presence of myringosclerosis and atrophy and audiometric thresholds above 1 kHz. No such correlation was observed with the presence of retraction pockets. In the BZO group middle ear admittance was observed significantly higher than in control group. The absent ipsilateral stapedial reflex was observed in 10.8% ears in BZO group, 16.9% in AM and 33.3% in KR. No correlation was found between the parameters of tympanometric evaluation and results of extended-high-frequency audiometry. CONCLUSIONS: In ears treated because of SOM with ventilation tube insertion the middle ear compliance is higher. In the presence of myringosclerosis and atrophy higher audiometric thresholds are observed. Tympanic membrane abnormalities have been more clearly indicated by the absent ipsilateral stapedial reflex than tympanometry.

[Long term sequelae of otitis media with effusion during childhood]. / Trwale nastepstwa wysiekowego zapalenia ucha w wieku dzieciecym.

INTRODUCTION: Otitis media with effusion is the most common cause of hearing loss in children and myringotomy with tympanostomy tube insertion is recommended procedure to deal with the problem. The objective of the present study was to determine the results of treatment, incidence and prevalence of middle ear sequelae and hearing results among children with chronic otitis media with effusion who received standard treatment with tympanostomy tubes. MATERIAL AND METHODS: The group of 97 patients treated by tympanostomy tubes insertion in the years 1999-2001 was reevaluated after mean period of follow up 7.3 years. At the control examination videootoscopy and audiologic examinations were performed. Audiological assessment consisted of tympanometry and pure-tone thresholds of air and bone conduction. RESULTS: Recurrent otitis media with effusion requiring tube insertion occurred during follow up period in 23.7% of patients. At the control examination 16.5% of children had an ongoing otitis media or ventilation tube in place or tympanic membrane perforation. Most common tympanic membrane abnormality were focal atrophy (67.2% of ears) and myringosclerosis (39.5%) followed by retraction pockets of pars flaccida (29.9%) and tensa (9.6%). Mean pure-tone audiometric threshold were significantly higher in ears with tympanic membrane abnormality by the difference did not excide 5dB HL in ears with focal atrophy and myringosclerosis and 6.5 dB HL in ears with retraction pockets. CONCLUSIONS: As the percentage of recurrences after tympanostomy tubes insertion are not uncommon prolonged period of follow up of those children is recommended. Although ventilation tubes have proven very effective in improving hearing in short term, they have not proven effective in preventing long-term sequelae of tympanic membrane and some degree of hearing loss. The decision about surgical treatment should be taken cautiously taking into account the chance of spontaneous resolution.

[CD80 and CD86 coreceptors expression on lymphocytes B in hypertrophied adenoids in children with otitis media with effusion]. / Ekspresja koreceptorów CD80 i CD86 na limfocytach B w przeroslych migdalkach gardlowych u dzieci chorych na wysiekowe zapalenie ucha srodkowego.

AIM: The aim of this study was evaluation of the percentage and mean fluorescence intensity of lymphocytes B with expression CD80 and CD86 in hypertrophied adenoids in children with otitis media with effusion. MATERIAL AND METHODS: Expression coreceptors CD80 and CD86 on lymphocytes B in adenoid tissue were estimated by flow cytometry method. 15 children with otitis media with effusion and 15 children with hypertrophied adenoid were tested. RESULTS: This study showed significantly higher percentage of lymphocytes CD19+ CD80+ in children with otitis media with effusion (OME 1.42% +/- 0.91) than in comparative group with hypertrophied adenoids (AH 0.63% +/- 0.21). The receptor CD86 didn't show differences in expression on lymphocytes B between both examinated groups. CONCLUSION: Low expression of CD80 and CD86 indicates that local immunity response in hypertrophied adenoids in children with otitis media with effusion is weakened. It can contribute to the otitis media with effusion.

Chosen factors of T and B cell apoptosis in hypertrophic adenoid in children with otitis media with effusion.

Evaluation of lymphocyte homeostasis within the chronically inflamed adenoid, closely related to the functioning of the immune system, may have a role in qualifying children for adenoidectomy. Apoptosis is a major process maintaining balance between tonsillar lymphocytes. The Fas receptor and Bcl-2 protein family which show pro-apoptotic and anti-apoptotic actions are of particular significance in apoptosis induction. Adenoid excised due to hypertrophy with or without chronic otitis media with effusion was used as study material. Flow cytometry was used to assess the percentages of apoptotic lymphocytes and CD4(+), CD8(+), CD19(+) cells with CD95(+) antigen and Bcl-2 protein in the group of children who underwent adenoidectomy due to adenoid hypertrophy and accompanying otitis media with effusion. The percentages of CD4(+)Bcl-2(+), CD8(+)Bcl-2(+) and CD19(+)Bcl-2(+) lymphocytes in the group of children with adenoid hypertrophy and acute otitis media were lower as compared to the reference group. However, the percentages of CD4(+), CD8(+) and CD19(+) cells with CD95(+) antigen were higher in the study group comparing to the reference group. The tendency of reduced percentages of T and B lymphocytes with Bcl-2 expression and elevated percentages of these cells with CD95(+) expression within the adenoid may reflect local immunity disorders.

[Clinical and pathological features of cholesteatoma in young children]. / Cechy kliniczne i patologiczne przewleklego zapalenia ucha z perlakiem u malych dzieci.

INTRODUCTION: Children cholesteatoma have been shown to demonstrate aggressive growth with greater extension and higher rates residual and recurrent disease compared with adults, due to anatomic and physiologic differences. AIM: This study aimed to determine the clinical and pathological features of cholesteatoma in young children less than 7 years of age. MATERIAL AND METHODS: Retrospective review of a group of 23 children (24 ears) who had cholesteatoma surgery between 1996 and 2004. Patients ages ranged from 1.8 to 7 years, follow up from 3.5 to 12 years (mean 7.4). Medical history (previous otorrhea, hearing loss) and contralateral ear status was analyzed. The extent of cholesteatoma and ossicular destruction for attic, pars tensa I and pars tensa II type and congenital of cholesteatoma was graded using of Saleh and Mills classification. Residual and recurrent disease depending on type of cholesteatoma, extent of the disease, type of surgery and contalateral ear status was assessed. RESULTS: Otorrhea and hearing loos are the most common clinical symptoms observed in 90% and 59% children. 70% of treated children had otitis media with effusion and/or its sequel in contralateral ear. 60% of acquired cholesteatomas were classified as S3 and S4 extension during surgery with ossicular destruction in 46% of attic cholestatoma. 75% operated ears had a signs of mucosa inflammation during surgery. Residual cholesteatoma was observed in 38.5% ears with attic cholesteatoma and in 25% with pars tensa I cholesteatoma. CONCLUSION: Acquired cholesteatoma in young children are diagnosed in the advanced stages with ossicular chain destruction. High rate of recidivism after surgery, particularly in attic cholesteatomas is observed. Most of operated ears have symptoms of inflammation of middle era mucosa during surgery and otitis media with effusion in contralateral ear and because of that need long-term follow up and constant observation.

[Expression CD27 on T and B lymphocytes in hypertrophied adenoids at children with otitis media with effusion]. / Ekspresja receptora CD27 na limfocytach T i B w tkance przeroslych migdalków gardlowych u dzieci chorych na wysiekowe zapalenie ucha srodkowego.

INTRODUCTION: Adenoid has particular meaning to develop of immunological response to inflammations in upper respiratory inclusive middle ear. The mining of antigen CD27 on lymphocytes T and B in creation of memory cells is still unclear. AIM: CD27 on lymphocytes T and B has a crucial role in development of immune response against inflammatory state. Aim of this study was evaluation functions of lymhocytes with expression CD27 in hypertrophied adenoid in children with otitis media with effusion. MATERIAL AND METHODS: [corrected] We tested 24 children in examine group with hypertrophied adenoid and otitis media with effusion (OME), and 29 children in comparative group only with hypertrophied adenoid (HA). We also divided both groups into two groups, above 5 and over 5 years old. We made the research by flow cytometry method. We used anty-CD19, anty-CD3, anty-CD5 and anty-CD27 monoclonal antibodies to examinations. RESULTS: We showed the lowest percentage lymphocytes B CD5+ with expression of CD27 in subgroup older children with otitis media with effusion (O.M.E. 22.43 +/- 2.66%) in compartative to younger children in the same group (15.96 +/- 5.31%; p < 0.005) and to older comparative subgroup (H.A. 15.50 +/- 8.03; p < 0.001). Percentages of B CD5 cells with expression of CD27 was statistical lower in examinated group (7.25 +/- 3.81) than in comparative group (H.A. 16.26 +/- 5.82; p < 0.04). In group with hypertrophied adenoid showed higher percentages of CD5-CD27+CD19+ cells both in younger and older supgroup than in equivalent supgroups with O.M.E. CONCLUSIONS: Iteraction of CD27-CD70 regulates many functions of T and B lymphocytes. It seems that disorders humoral immunologucal answer in hypertrophied adenoids are supported to develop of otitis media with effusion.

[Expression of chosen chemokine receptors on the Th lymphocytes in children with hypertrophied adenoids with otitis media with effusion]. / Ekspresja wybranych receptorów chemokin na limfocytach Th przeroslych migdalków gardlowych w wysiekowym zapaleniu ucha srodkowego.

INTRODUCTION: Chemokine receptors play a crucial role in the recruitment of leucocytes into inflamed tissue from secondary lymphoid organs. AIM OF THIS STUDY: was evaluation of the percentage Th (CD4+) lymphocytes with expression of chemokine receptors: CCR3, CCR4, and CCR5 in hypertrophied adenois tissue in children with otitis media with effusion. MATERIAL AND METHODS: 36 children with otitis media with effusion and 25 children with hypertrophied adenoids were tested. Expression of chemokine receptors CCR3, CCR4 and CCR5 on T CD4 lymphocytes of hypertrophied adenoid's tissue was estimated by flow cytometry method. RESULTS: Average percentage of T CD4+ lymphocytes with expression CCR4 in hypertrophied adenoid in children group with otitis media with effusion (OME = 64.11%) was significantly higher than in comparative group with hypertrophied adenoid (HA = 75.05%, p < 0.04). We showed statistically lower percentage of CD3+CD4+ cells with expression CCR3 in examinated group (OME = 47.19%) than HA group (62.66%; p < 0.003). CONCLUSION: Results suggests that intensify of humoral immunological answer in hypertrophied adenoid depends on Th2 lymphocytes with expression CCR3 and CCR4 chemokine receptors in course of inflammatory state in middle ear.