Influence of the chemical composition of leachates on the results of ecotoxicity tests for different slag types.

In this study, four ecotoxicological tests on Vibrio fischeri bacteria, Sinapis alba L. (white mustard), Daphnia magna S. (daphnia's) and earthworms were performed for three types of aqueous slag (ladle, blast furnace and converter) leachates with two-grain sizes (<4 mm, <10 mm). Concentrations of toxic elements and concentrations of Cr(VI), Ca, Na, Al, and other ions were determined. The raw slags were analyzed using X-ray fluorescence spectroscopy (XRFS), and major substances were determined by X-ray powder diffraction (XRD). The aqueous slag leachates passed ecotoxicological tests and met the required criteria, showing no toxicity to Vibrio fischeri and complying with white mustard test criteria. According to the results of the ecotoxicity tests with daphnia, the blast furnace slag samples were not ecotoxic, while two other slag samples were found to be entirely compliant. Characterization of the slags showed that the effect of element/ion leachability and slag grain size is essential. Biplot principal component analysis (PCA) showed that grain size does not significantly affect the separation of individuals on the plane. A positive correlation on toxicity was found with pH, conductivity, calcium content, dissolved content, salinity and fluoride concentration, whereas a negative correlation was found with magnesium concentration, dissolved organic carbon and potassium concentration. The effective concentration at 50% inhibition (EC50) value for Vibrio fischeri correlated with the first dimension of bivariate assessment. In summary, it was found that the investigated slags can be effectively reused as they comply with regulations and do not endanger the environment.

HPV and RAD51 as Prognostic Factors for Survival in Inoperable Oral and Oropharyngeal Cancer in Patients Unfit for Chemotherapy Treated with Hyperfractionated Radiotherapy.

INTRODUCTION: The incidence of advanced oral cavity and oropharyngeal cancers is generally high. Treatment outcomes for patients, especially those unfit for comprehensive cancer treatment, are unsatisfactory. Therefore, the search for factors to predict response to treatment and increase overall survival is underway. OBJECTIVE: This study aimed to analyze the presence of 32 HPV genotypes in tumor samples of 34 patients and the effect of HPV status and RAD51 on overall survival. METHOD: Tumor samples of 34 patients with locally advanced oropharyngeal or oral cavity cancer treated with accelerated radiotherapy in monotherapy were analyzed using reverse hybridization and immunohistochemistry for the presence of HPV and RAD51. Its effect on overall survival was examined. RESULTS: Only two types of HPV were identified-HPV 16 (dominant) and HPV 66 (two samples). The HPV positivity was associated with a borderline insignificant improvement in 2-year (p = 0.083), 5-year (p = 0.159), and overall survival (p = 0.083). Similarly, the RAD51 overexpression was associated with borderline insignificant improvement in 2-year (p = 0.083) and 5-year (p = 0.159) survival. CONCLUSION: We found no statistically significant differences but detected trends toward improvement in the survival of HPV-positive and RAD51 overexpressing patients unfit for surgical treatment or chemotherapy treated with hyperfractionated radiotherapy. The trends, however, indicate that in a larger group of patients, the effects of these two parameters would likely be statistically significant.

Two Bonebridge bone conduction hearing implant generations: audiological benefit and quality of hearing in children.

PURPOSE: The study aimed to evaluate audiological benefits, quality of hearing and safety of two Bonebridge generation: BCI601 and BCI602 (MED-EL, Innsbruck, Austria) in children. METHODS: Twelve children were implanted: five BCI601 and seven BCI602 comprising of ten conductive hearing loss, and two single sided deaf SSD subjects. Audiological outcomes tested were sound field audiometry, functional gain, speech recognition threshold (SRT50), speech recognition in noise (SPRINT) and localisation abilities. Subjective measures were Speech, Spatial and Qualities of Hearing Scale (SSQ12). RESULTS: The mean FG with the BCI601 was 25.0 dB and with the BCI602 28.0 dB. The benefit in SRT50 was 23.2 dB and 33.8 dB, respectively. The mean benefit in SPRINT was 15% and 6.7% and the localisation ability improved from 33.3° to 16° and from 26.2° to 17.6°, respectively. The two SSD subjects reported a FG of 17 dB, a benefit in SRT50 of 22.5 and a benefit in SPRINT of 20%. Subjective outcomes improved significantly and even exceeded the values of their age-and sex matched normal hearing peers. One revision was reported: a retroauricular emphysema above the implant occurred 12 months post-OP, it was resolved operatively with the implant still being functional. CONCLUSION: The pediatric cohort reports significant audiological benefit, even exceeding that of the age- and sex matched control. The combination of the high safety and audiological benefit makes the Bonebridge a comfortable and effective option in hearing rehabilitation in children.

Chlorophyll Content in Two Medicinal Plant Species Following Nano-TiO2 Exposure.

Chlorophyll content in leaves is a convenient indicator of the physiological state of the plant following exposure to a stressor. In the present study, a pot experiment was carried out using two medicinal plant species - peppermint (Mentha X piperita L.) and common sage (Salvia officinalis L.) to determine the link between the chlorophyll content in the plant leaves and the exposure of the plants to nano-TiO2 either through the leaves (in suspension) or through the root system (in soil). Following the exposure, the shoots were analysed for the contents of Ti, Al, Ca, K, Mg, Mn, Na and chlorophyll. Significant decrease in chlorophyll content was observed in all but one of the nano-TiO2 treatments, the differences in the determined element content were mostly species-dependent: Ti exposure was found to be related to the decrease in the chlorophyll and Mn content in both species as well as to the decrease in Mg content in common sage.

The role of metabolic syndrome in the induction of chronic pancreatitis after a first attack of acute pancreatitis - multicenter trial.

BACKGROUND: Metabolic syndrome is a serious societal problem worldwide. In the Czech Republic more than 30% of the adult population are sufferers. The role of recurrent acute pancreatitis in the induction of chronic pancreatitis, following the socalled „mechanistic definition“ of chronic pancreatitis, has been unequivocally confirmed. However, there are a number of factors that may contribute to the development of chronic pancreatitis. The first aim of the study was to determine whether the metabolic syndrome may affect the development of chronic pancreatitis. The second question we asked ourselves was whether even one acute attack of pancreatitis could be an inductive factor in chronic pancreatitis. METHODS: Based on data obtained retrospectively from a total of 264 people diagnosed with chronic pancreatitis in 4 centers, a total of 59 people (22.3%) diagnosed within 36 months of a first attack of acute pancreatitis was obtained. Etiologies of either genetically induced pancreatitis or autoimmune pancreatitis were excluded. Diagnostics to identify the presence of metabolic syndrome were run on the 59 persons so obtained using the criteria from the socalled „harmonized“ definition of 2009 (obesity, arterial hypertension, hypertriglyceridemia, type 2 diabetes mellitus and a decreased level of HDL cholesterol). RESULTS: Comparing the findings of the individual components of metabolic syndrome in persons with chronic pancreatitis after a 1st attack of acute pancreatitis with the metabolic syndrome and in persons with chronic pancreatitis after the 1st attack of acute pancreatitis but without metabolic syndrome, a statistically significant difference in obesity was found (82.5% vs. 28.5%), hypertriglyceridemia (82.3% vs 17.8%) and arterial hypertension (70.5% vs 21.4%). The interval during which chronic pancreatitis occurred after acute pancreatitis averaged 12 months (10-14 months) in subjects with metabolic syndrome, whereas in the group without metabolic syndrome the interval was longer, 20 months (16-29 months). CONCLUSION: Our results show that even one attack of acute pancreatitis (regardless of etiology) can be an inductive factor in chronic pancreatitis. The presence of metabolic syndrome can accelerate the development of chronic pancreatitis after one has had acute pancreatitis.

Ti and Zn Content in Moss Shoots After Exposure to TiO2 and ZnO Nanoparticles: Biomonitoring Possibilities.

To assess the uptake of nanoparticles by moss shoots and the possibility of biomonitoring the moss of nanoparticle pollution, two moss species frequently used in biomonitoring surveys [Hylocomium splendens (Hedw.) Schimp. and Pleurozium schreberi (Brid.) Mitt.] were repeatedly exposed to known concentrations of either nano-TiO2 or nano-ZnO suspensions. The interspecies differences were assessed by exposing both the species to 1 g L-1 nano-ZnO suspension, H. splendens samples were also exposed to either 0.1 g L-1 or 1 g L-1 suspension of nano TiO2. The exposed samples were analysed for Zn or Ti content using Inductively Coupled Plasma-Atomic Emission Spectroscopy. Both species showed a similar accumulation pattern, H. splendens being a slightly better accumulator. The washing suggests that Ti successfully penetrated the interior of the gametophyte. Since the relationship between the exposure and accumulation is linear, moss biomonitoring is, hereby, considered to be a viable, novel technique in nanoparticle pollution assessment.

The first evidence of microplastic occurrence in mine water: The largest black coal mining area in the Czech Republic.

Climate change is creating new challenges for water supply worldwide, making the search for new sources of water vital. As mine water could serve as a potential source, this study investigated the presence of microplastics in water from terminated deep mines in the largest coal basin in the Czech Republic, as well as in water from nearby shallow wells. The particles found were analyzed for size, polymer composition, color and morphology using the ImageJ tool, infrared spectroscopy with Fourier transform (FTIR) and an optical stereomicroscope with a digital camera. Microplastics were detected in all tested sites. Their range accounted for 2.5-17.5 items/L for mine water samples and 2.5-20 items/L for well samples, with fibers being the dominant type. The average width of particles from mine water and wells amounted to 58 µm; 71 µm, length to 655 µm; 501 µm and area to 22,067 µm2; 28,613 µm2, respectively. Blue color was prevalent, among materials, in both cases, plastic coated paper was found dominant to Polyethylene terephthalate (PET), Polyester (PES), Tetrafluoroethylene-perfluoro (Propyl Vinyl Ether) - Copolymer (TFE-PPVE), and polypropylene (PP). The research provides the first evidence of microplastics' presence in underground waters from deep mines and shallow wells in the same area. The data suggest that it is almost impossible to find underground water sources free of microplastic contamination. In this context, atmospheric contamination from mine ventilation and infiltration through terminated mines were identified as potential sources, while infiltration through soil and rock formations is unlikely given the geological composition. The results of this study can serve as a relevant basis for further research on microplastics in mine waters. Additionally, the conclusions can advance the development in remediation technologies of microplastics from deep underground waters and their implementation in practice, particularly in light of upcoming legislation.

Alteration of Hordeum vulgare and Sinapis alba germination and early growth in response to airborne low-metallic automotive brake wear debris.

Road transportation significantly contributes to environmental pollution, both in terms of exhaust and non-exhaust (brake wear) emissions. As was proven, brake wear debris is released in a wide variety of sizes, shapes, and compositions. Although studies confirming the possible adverse health and environmental impact of brake wear debris were published, there is no standardized methodology for their toxicity testing, and most studies focus only on one type of brake pad and/or one test. The lack of methodology is also related to the very small amount of material released during the laboratory testing. For these reasons, this study deals with the mixture of airborne brake wear debris from several commonly used low-metallic brake pads collected following the dynamometer testing. The mixture was chosen for better simulation of the actual state in the environment and to collect a sufficient amount of particles for thorough characterization (SEM, XRPD, XRF, chromatography, and particle size distribution) and phytotoxicity testing. The particle size distribution measurement revealed a wide range of particle sizes from nanometers to hundreds of nanometers, elemental and phase analysis determined the standard elements and compounds used in the brake pad formulation. The Hordeum vulgare and Sinapis alba were chosen as representatives of monocotyledonous and dicotyledonous plants. The germination was not significantly affected by the suspension of brake wear debris; however, the root elongation was negatively influenced in both cases. Sinapis alba (IC50 = 23.13 g L-1) was more affected than Hordeum vulgare (IC50 was not found in the studied concentration range) the growth of which was even slightly stimulated in the lowest concentrations of brake wear debris. The plant biomass was also negatively affected in the case of Sinapis alba, where the IC50 values of wet and dry roots were determined to be 44.83 g L-1 and 86.86 g L-1, respectively.

Simple method for quantification of metal-based particles in biopsy samples of patients with long bone implants - Pilot study.

The presence of particles fixed in tissue samples due to implant degradation or disintegration plays an important role in post-operative complications. The ability to determine the size, shape, chemical composition and, above all, the number of these particles can be used in many areas of medicine. This study presents a novel, simple metal-based particle detection method using scanning electron microscopy with energy dispersive spectrometer (SEM-EDS). The presence of metal particles in biopsy specimens from long bone nail-fixated implants (10 patients with titanium steel nails and 10 patients with stainless steel nails) was studied. The samples were analysed using automated area analysis based on image binarization and brightness to 255 grayscale. The results were supplemented with histological data and statistically analysed. The method based on the software used was found to be accurate and easy to use and, thus, appears to be very suitable for particle detection in similar samples.

Microplastics and additives in patients with preterm birth: The first evidence of their presence in both human amniotic fluid and placenta.

Considering the well documented, almost ubiquitous nature of microplastics in different environments, the exposure of pregnant women to microplastics may pose risks to the unborn children. The study focused on investigating the presence of microplastics in amniotic fluid and placenta, and brings the first evidence of the simultaneous presence of microplastics and additives in both human amniotic fluid and placentas. In total, 20 samples of amniotic fluid and placenta from 10 patients were analyzed for the presence of microplastics and plastic additives by Fourier transform infrared spectroscopy - attenuated total reflectance (FTIR-ATR) after alkaline digestion with KOH. In 9 out of 10 patients, microplastics or additives were found in amniotic fluid, placenta, or both. Specifically, 44 particles of microplastics and polymer additives were identified in all samples. Chlorinated Polyethylene (CPE) and Calcium zinc PVC Stabilizer with particle sizes between 10 and 50 µm prevailed. Although all women involved in this study, who provided placenta and amniotic fluid samples, experienced physiological, singleton pregnancies complicated with preterm prelabour rupture of membranes (PPROM), it is too early to draw any conclusions and more research is needed.

Microbiota Diversity in Non-Small Cell Lung Cancer Gut and Mouth Cavity Microbiota Diversity in Non-Small Cell Lung Cancer Patients.

Lung malignancies have a substantial impact on cancer incidence and mortality worldwide. Even though many factors involved in the development of the disease are known, many questions remain unanswered. Previous studies suggest that the intestinal microbiota may have a role in developing malignant diseases. According to some findings, the microbiota has proven to be a key modulator of carcinogenic processes and the immune response against cancer cells, potentially influencing the effectiveness of immunotherapy. In our study, we characterized culturable microorganisms associated with non-small cell lung cancer (NSCLC) that can be recovered from rectal swabs and mouthwash. In addition, we also explored differences in the culturable microbiota with two main types of NSCLC - adenocarcinoma (ADC) and squamous cell carcinoma (SCC). With 141 patients included in the study (86 ADC and 55 SCC cases), a significant difference was observed between the two types in seven bacterial species (Collinsella, Corynebacterium, Klebsiella, Lactobacillus, Neisseria, Rothia, and Streptococcus), including the site of origin. The relationship between microbial dysbiosis and lung cancer is poorly understood; future research could shed light on the links between gut microbiota and lung cancer development.

Enhancing oral squamous cell carcinoma prediction: the prognostic power of the worst pattern of invasion and the limited impact of molecular resection margins.

Objective: Oral squamous cell carcinoma (OSCC) originates from the mucosal lining of the oral cavity. Almost half of newly diagnosed cases are classified as advanced stage IV disease, which makes resection difficult. In this study, we investigated the pathological features and mutation profiles of tumor margins in OSCC. Methods: We performed hierarchical clustering of principal components to identify distinct patterns of tumor growth and their association with patient prognosis. We also used next-generation sequencing to analyze somatic mutations in tumor and marginal tissue samples. Results: Our analyses uncovered that the grade of worst pattern of invasion (WPOI) is strongly associated with depth of invasion and patient survival in multivariable analysis. Mutations were primarily detected in the DNA isolated from tumors, but several mutations were also identified in marginal tissue. In total, we uncovered 29 mutated genes, mainly tumor suppressor genes involved in DNA repair including BRCA genes; however none of these mutations significantly correlated with a higher chance of relapse in our medium-size cohort. Some resection margins that appeared histologically normal harbored tumorigenic mutations in TP53 and CDKN2A genes. Conclusion: Even histologically normal margins may contain molecular alterations that are not detectable by conventional histopathological methods, but NCCN classification system still outperforms other methods in the prediction of the probability of disease relapse.

Traffic pollution tracers in the lymphatic system tissue of children-possible link to chronic tonsillitis development: pilot study.

The origin of metal elements in tonsillar tissues of children patients was investigated in order to establish the link between environmental pollution and tonsilitis diagnosis. The children lived either in a polluted, industrial region (Moravian-Silesian Region, North-East Czechia) or a clean, predominantly agricultural region (South Bohemian Region, South-East Czechia). Simultaneously, the distance of the address of the patients to the closest main road was assessed. The elemental content of the present particulate matter in the tonsil samples, coming from indicated tonsillectomies, was analysed using scanning electron microscopy equipped with an X-ray microprobe. No significant association of the elemental composition and the region was found using both bivariate and multivariate methods; however, an association with the distance to the closest main road was found. The study presents a novel assessment of the tonsillar tissues and supports the link between environmental factors (traffic-related particulate pollution) and the onset of chronic, idiopathic diseases in children, which has been debated recently.

Magnetically Modified Biosorbent for Rapid Beryllium Elimination from the Aqueous Environment.

Although both beryllium and its compounds display high toxicity, little attention has been focused on the removal of beryllium from wastewaters. In this research, magnetically modified biochar obtained from poor-quality wheat with two distinct FexOy contents was studied as a sorbent for the elimination of beryllium from an aqueous solution. The determined elimination efficiency was higher than 80% in both prepared composites, and the presence of FexOy did not affect the sorption properties. The experimental qmax values were determined to be 1.44 mg/g for original biochar and biochar with lower content of iron and 1.45 mg/g for the biochar with higher iron content. The optimum pH values favorable for sorption were determined to be 6. After the sorption procedure, the sorbent was still magnetically active enough to be removed from the solution by a magnet. Using magnetically modified sorbents proved to be an easy to apply, low-cost, and effective technique.

Identification of the phase composition of solid microparticles in the nasal mucosa of patients with chronic hypertrophic rhinitis using Raman microspectroscopy.

Solid particles, predominantly in micron and submicron sizes, have repeatedly been observed as a threat to a human health unique compared to the other textures of the same materials. In this work, the hypothesis the solid metal-based particles play a role in the pathogenesis of chronic hypertrophic rhinitis was investigated in patients who had not responded positively to medication. In the group of 40 randomly selected patients indicated for surgical mucotomy, the presence of solid micro- and submicron particles present in their nasal mucosa was assessed. For comparison, a set of 13 reference samples from patients without diagnosed chronic hypertrophic rhinitis was evaluated. The analysis was performed using Raman microspectroscopy. The advantage of this method is the direct identification of compounds. The main detected compounds in the mucosa samples of patients with chronic hypertrophic rhinitis were TiO2, carbon-based compounds, CaCO3, Ca(Fe, Mg, Mn)(CO3)2 MgCO3, Fe2O3, BaSO4, FeCO3 and compounds of Al and Si, all of which may pose a health risk to a living organism. In the reference samples, only TiO2 and amorphous carbon were found. In the control group mucosa, a significantly lower presence of most of the assessed compounds was found despite the longer time they had to accumulate them due to their higher mean age. Identification and characterisation of such chemicals compounds in a living organism could contribute to the overall picture of the health of the individual and lead to a better understanding of the possible causes not only in the chronic hypertrophic rhinitis, but also in other mucosal and idiopathic diseases.

Cardiac papillary fibroelastomas: A 10-year single-center surgical experience and long-term echocardiographic follow-up study.

AIMS: Limited contemporary data are available on the clinical and echocardiographic outcomes after surgery for cardiac papillary fibroelastoma (CPF). The aim of this study was to review the clinical manifestations, pathological characteristics, surgical management, and prognoses of patients with histologically verified CPF, who underwent surgery at our cardiac surgery center from 2008 to 2018. METHODS AND RESULTS: Twelve patients of median age 62 years (28 to 77 years) were treated. Embolic stroke or transient ischemic attack (five patients, 42%) were the only CPF clinical manifestations. Eleven (92%) tumors were localized on the valves, with the aortic valve being the most common tumor site (seven patients; 58%). Multiple factor analysis revealed no independent predictor of CPF-related embolization. Simple shave tumor excision was sufficient in most patients (10 patients, 83%). No operative or tumor-related late mortality during the median follow-up period of 4.7 years (1.1 to 10.2 years) was recorded. Asymptomatic metachronous valve tumor recurrence (in a location different from that of the original tumor) was revealed in two patients (17%) by transesophageal echocardiography (TEE), not detected by transthoracic echocardiography (TTE). One of these two patients underwent repeated surgery for CPF but later suffered a recurrent embolic stroke, due to another tumor recurrence. CONCLUSION: CPF can be safely and effectively treated surgically. TEE is superior to the TTE option in CPF post-operative recurrence detection. There is a clear need for a prospective study to determine criteria for embolization risk stratification and optimum management in patients with CPF.

Moss biomonitoring and air pollution modelling on a regional scale: delayed reflection of industrial pollution in moss in a heavily polluted region?

A passive biomonitoring survey using terrestrial mosses was performed in a heavily polluted industrial region on the border between Czechia and Poland in a regular grid of 41 sampling points. The concentrations of 38 elements were determined in the moss samples, using Neutron Activation Analysis (NAA). Simultaneously, air pollution modelling was performed using the Czech reference methodology Symos'97 for the year of the sampling (2015) and 3 years prior (2012) in order to compare the results of both the approaches and evaluate the credibility of the moss biomonitoring method. The NAA results were transformed according to the principles of compositional data analysis and assessed using hierarchical clustering on principal components. The resulting clusters were compared with the results of air pollution modelling using one-way analysis of variance. The association of determined clusters with the pollution from industrial sources was confirmed only for the results of the 2012 modelling. This validates the complementarity of the air pollution modelling and the moss biomonitoring, ascertains the moss biomonitoring as a valid method for long-term pollution assessment and confirms one of the fundamentals of moss biomonitoring, the reflection of the atmospheric conditions prevailing in the period before the sampling.

Multivariate assessment of atmospheric deposition studies in Bulgaria based on moss biomonitors: trends between the 2005/2006 and 2015/2016 surveys.

This study aims to investigate the changes in atmospheric deposition trends in Bulgaria, studied using the moss biomonitoring technique since 1995. For the first time, a paired (site-wise) comparison was performed after a critical review of the sampling networks and adjusting for location, the distance between the sampling points, and moss species. Data from the 2005/2006 and 2015/2016 moss surveys were chosen as instrumental neutron activation analysis was employed in both to determine the content of 34 elements (Al, As, Ba, Br, Са, Ce, Cl, Со, Cr, Cs, Fe, Hf, I, K, La, Mn, Na, Nd, Ni, Rb, Sb, Sc, Se, Sr, Ta, Tb, Th, Ti, Tm, U, V, W, Yb, Zn). In addition, Cd, Cu, and Pb were determined using complementary analytical methods: inductively coupled plasma atomic emission spectroscopy in 2015/2016 and atomic absorption spectroscopy in 2005/2006. For the subset of 57 routinely sampled locations in Bulgaria, hierarchical clustering on principal components and multiple factor analysis (MFA) were applied to assess the spatial trends in the 10 years elapsed between the surveys, as well as to characterise the origin of the determined elements. Elevation and distance between the sampling points were used as additional variables in the multiple factor analysis plane to ascertain their effect on the overall variance in the datasets. Distribution maps were constructed to illustrate the deposition patterns for the pollutant Pb. The results were consistent with decreased industrial output in the country, increased coal combustion and transport pollution, and construction of roads.

Metal particles in mucus and hypertrophic tissue of the inferior nasal turbinates from the human upper respiratory tract.

Mucosal surfaces are the first mechanical barrier preventing the entry of foreign particles into the organism. The study addresses the detection and analysis of metal-based solid particles in cytological mucus samples from the surface of human hypertrophic tissue in the inferior nasal turbinates in patients diagnosed with chronic rhinitis. Solid particles were characterized by scanning electron microscopy and Raman microspectroscopy; all the biological samples were also subjected to vibration magnetometry. Since the upper airways are the first part of the respiratory tract, which is exposed to inhaled particles, it can be assumed that inhaled particles may be partially deposited in this region. Scanning electron microscopy revealed the presence of metal-based solid particles/clusters in the majority of the analysed cytological mucus samples and also in hypertrophic tissues; in all groups, the particles were of submicron size. Raman microspectroscopy detected the presence of particles/clusters based on amorphous carbon, graphite, calcium carbonate, anatase and barite only in the hypertrophic tissue. The obtained results show that the composition of some of the solid particles (i.e. Ba, Zn, Fe and Ti) detected in the mucus from the surface of the hypertrophic tissues resembled the particles found in the hypertrophic tissue itself. It can be assumed that after the capture of the inhaled particles by the mucus, they penetrate into the deeper layers of tissue.

Immunohistochemical analysis of retraction pocket pars tensa of tympanic membrane in children.

AIMS: Immunohistochemical analysis of retraction pocket pars tensa of tympanic membrane in children. Identification of signs typical for cholesteatoma and support of retraction theory of cholesteatoma. STUDY DESIGN: a prospective study analysing 31 surgically removed retraction pockets. DEPARTMENT: University Hospital, Children's Medical Centre Methods: Retraction pockets processed by a standard process for immunohistochemical analysis. The observed findings were specified using antibodies CD45 LCA (leukocyte common antigen), CD31 (platelet endothelial cell adhesion molecule), D2-40 (marker of lymphatic endothelium), MMP9 (marker of degradation of connective tissue extracellular matrix) and Ki67 (cellular marker of proliferation). RESULTS: All observed parameters except for MMP9 had a significantly higher incidence in retraction pocket stage III compared to stage II according to Charachon. CONCLUSION: We described immunohistochemical signs of retraction pocket pars tensa of tympanic membrane in children resulting in cholesteatoma. All the observed signs occur in the structure of matrix and perimatrix of cholesteatoma. A significantly higher incidence of all observed parameters except from MMP9 was proved in retraction pocket stage III, unlike in stage II. This observation proves the fact that retraction pocket is a progressive disease and is a procholesteatoma stage.