Immunohistochemical Identification and Assessment of the Location of Immunoproteasome Subunits LMP2 and LMP7 in Acquired Cholesteatoma.

Cholesteatoma, accompanied by chronic inflammatory response, is characterized by invasive growth and osteolytic activity. As specific proteasome isoforms, the immunoproteasomes serve as an important modulator of inflammatory responses. The aim of the present study was to determine the biological activity of cholesteatoma through the analysis of the expression and localization of immunoproteasome subunits of low molecule weight protein (LMP) 2 and LMP7. Cholesteatoma specimens were obtained from 15 adults who underwent ear surgery due to acquired attic cholesteatoma. Normal skin specimens were taken from retro-auricular skin incisions from the same patients. The specimens were stained with anti-LMP7 antibody, using immunohistochemistry techniques based on the binding of biotinylated secondary antibody with the enzyme-labeled streptavidin and the Envision FLEX system. In all specimens of cholesteatoma, the immunohistochemical reaction with the antibody against the LMP2 was positive, in both the cytoplasm of the cholesteatoma matrix and the perimatrix. A negative reaction with anti-LMP2 was observed in the cytoplasm and nuclei of control skin cells. A positive nuclear and cytoplasmic immunohistochemical reaction with anti-LMP7 has been demonstrated in numerous cells, in both the matrix and perimatrix of cholesteatoma. We present evidence of the presence of expressions of LMP2 and LMP7 within cholesteatoma tissue. Our results might bring new information concerning immunoproteasome-dependent pathophysiologic mechanisms in cholesteatoma.

Cholesteatoma Definition and Classification: A Literature Review.

Cholesteatoma is a serious otolaryngologic condition that to date remains an important problem and poses a challenge to otolaryngologists around the world. To improve the approach pertaining to the diagnosis and management of middle ear cholesteatoma, clear, clinically applicable, and useful definition and classification of cholesteatoma are required. This review aimed to evaluate the current and most accepted descriptions and opinions concerning cholesteatoma. A review of the literature concerning different definitions and classifications of cholesteatoma was used in the preparation of the Cholesteatoma Guidelines, a project implemented by the European Academy of Otology - Neuro-otology.

Quality of life and hearing after cochlear implant placement in patients over 60 years of age.

OBJECTIVE: Multicenter evaluation of the quality of life and quality of hearing after Nucleus® cochlear implant placement in patients over 60 years of age. REFERENCE AND METHOD: Evaluation was performed in patients receiving cochlear implants after the age of 60 years as a part of the Cochlear-Implanted Recipient Observational Study (Cochlear-IROS). This study is a prospective, international and long-term assessment which enables observation of recipients for up to three years after implantation. Data regarding subjective evaluation of the quality of life and quality of hearing were gathered before the first switch-on of the sound processor and one year afterwards. Standardized questionnaires were used in this evaluation, including Health Utility Index (HUI mk. III) and Speech, Spatial and Qualities of Hearing (SSQ) Scale. Data were also gathered regarding the aetiology of hearing loss, hearing aid usage, tinnitus and vertigo, as well as on the telephone usage and the professional status of recipients. RESULTS: Included in the evaluation, were 20 subjects who were over 60 years old at the moment of the cochlear implant surgery. The study group consisted of 12 men and 8 female patients. The average age of CI recipients at the moment of implantation was 67.8 years (min. 60, max. 80 years). The SSQ questionnaire outcomes regarding self-assessment in the field of ability to hear in everyday situations one year after the surgery indicate that speech understanding increased by 180%, spatial hearing increased by 135 % and quality of hearing increased by 98%. Overall quality of life before the first sound processor switch-on as assessed using the HUI questionnaire was at the level of 0.38 (on 0-1 scale, where 0 equals death, and 1 equals full health). One year after the implantation, this assessment increased by 33% (up to 0.5 on the scale). CONCLUSION: The effectiveness of the cochlear implantation in patients with severe hearing loss after 60 years of age with respect to the quality of life and hearing was confirmed. Statistically significant improvement was demonstrated in the self-assessment of patients in relation to the speech understanding, spatial hearing and quality of hearing, as well as quality of life.

Consensus-Based Recommendations on the Definition and Classification of Cholesteatoma.

Development of a consensus on the definitions and classification of cholesteatoma is essential for scientific community to exchange information on clinical studies and compare their outcomes. The aim of the study is to reach a consensus among members of the European Academy of Otology and Neurotology (EAONO) regarding the definitions and classification related to cholesteatoma. A set of statements was developed by the authors utilizing the literature on the definition and classification of cholesteatoma. A questionnaire was sent to the members of the EAONO, inviting them to state if they are in agreement with each of the statements and if not, then to provide comments or suggestions for revision. Responses were evaluated and modified using online questionnaire and survey software based on the Delphi technique, a cyclical process of gathering information, summarizing, and re-submitting the revised statements to the same target population until a consensus is reached. Target agreement among the responders was set at a minimum of 80%, and the cycle of revision and re-submission of the statements were repeated until a consensus was reached on a majority of the statements. A steering group has been established to evaluate the results of the survey and worked via the process of cognitive debriefing. Out of 364 EAONO members, 123 responded to the first consensus cycle, 77 to the second cycle, and 53 to the third cycle. After three cycles, all statements concerning cholesteatoma definitions reached the target of 80% consensus. However, a consensus on the classification of cholesteatoma could not be achieved. The steering group excluded four statements of cholesteatoma definition and established a consensus on cholesteatoma classification. A consensus on cholesteatoma definitions was reached among the members of the EAONO. The final revision on consensus statements for cholesteatoma definition and classification has been made via the process of cognitive debriefing of the steering group.

The role of p21 and p53 proteins in congenital cholesteatoma.

OBJECTIVE: Immunoexpression analysis of p53 and p21 in congenital cholesteatoma with assessing their exact localization in cholesteatoma layers and the level of expression. P53 and p21 are apoptosis-related molecules that regulate cell cycle. These markers have been not completely evaluated in congenital cholesteatoma, and the role of apoptosis in congenital cholesteatoma is also not completely understood. METHODS: Congenital cholesteatoma samples, a study group (n = 13) and normal auditory meatal skin, a control group (n = 12) from patients who underwent surgery for cholesteatoma were included in the study. Acquired cholesteatoma samples were used as a comparable group (n = 12). Tissue sections were investigated with the immunohistochemistry technique based on binding of biotinylated secondary antibody with the enzyme-labeled streptavidin with using appropriate primary antibodies. Cells with immunoexpression of analyzed antigens: p53 and p21 were defined as antigen positive. In each section, cells were counted, and the percentage of positive cells was determined. The level of significance was set at p < 0.05. RESULTS: The mean percentage of p21-positive was statistically significant higher in congenital cholesteatoma than in the control group (p < 0.05). There was no meaningful difference between congenital and acquired cholesteatoma with respect to p21 expression. There was significant difference between congenital and acquired cholesteatoma regarding p53. CONCLUSION: Up-regulation of p21 protein is expected to play a significant role in CC development. Apoptosis is an important process in the pathogenesis of congenital cholesteatoma. It seems reasonable to perform studies on apoptosis in congenital cholesteatoma in the prospective way taking into consideration more specimens.

Selected surgical managements in snoring and obstructive sleep apnea patients.

BACKGROUND: The diagnostic process and the surgical procedures in patients with snoring and obstructive sleep apnea syndrome (OSAS) are crucial. The aim of this study was to assess the efficacy of surgical treatment in snoring and OSAS patients. MATERIAL/METHODS: A precise laryngological examination and screening polysomnography (Poly-Mesam) were performed in all patients with mild, moderate and severe OSAS before and 6 months after surgery. The patients completed questionnaires concerning their complaints. We included patients qualified to septoplasty, laser-assisted uvulopalatoplasty (LAUP), uvulopalatopharyngoplasty (UPPP) and radiofrequency-induced thermotherapy of the tongue base (RITT). Outcome evaluation of surgery was performed on the basis of data received from follow-up laryngological examinations, selected parameters obtained from the Poly-Mesam test and follow-up questionnaires. RESULTS: In most cases we observed improvement, defined as decreasing some sleep parameters, such as a respiratory disturbance index (RDI), by more than 50%, decreasing the loudness of snoring, decreasing the number of hypopneas, and obtaining better blood saturation values. After UPPP we noticed changes in retropalatal space, soft palate dimensions and uvula-posterior pharyngeal wall distance. In the postoperative period we did not observe severe complications. In some cases we found short-lived palatal deficiency after UPPP. Patients after RITT experienced discomfort and throat pain lasting from 2 to 4 days. In 2 patients we observed swelling of the tongue base, which decreased after few days. CONCLUSIONS: Surgery in OSAS contributes to normalization of some sleep parameters. The majority of patients experienced improvement after surgery.

Cytology of nasal mucosa, olfactometry and rhinomanometry in patients after CO2 laser mucotomy in inferior turbinate hypertrophy.

To evaluate the cytology of nasal mucosa and sense of smell and nasal patency in patients underwent carbon dioxide laser turbinoplasty (CO2 laser mucotomy) due to chronic nasal hypertrophy. 46 patients with inferior turbinate hypertrophy underwent complete laryngological examination, anterior rhinomanometry, olfactory measurements and cytology of nasal mucous which were performed before and 3 months after CO2 laser mucotomy. Laser mucotomy was performed under local anesthesia. Cytograms revealed significant changes in cell proportion before and after the surgery. Goblet cells predominated in nasal smears before the laser mucotomy. An average percentage of eosinophils in evaluated cytograms before the surgery was 2.1%. Three months after laser mucotomy we observed decrease in goblet cells proportion (the mean range of goblet cells was 16.4%) in nasal cytology. We have also observed improvement in olfactory function, however only in 7 patients (20.6%). The mean value of total nasal airway resistance (NAR) before treatment was 0.98+/-0.24 Pa/cm3/s at 75 Pa. Rhinomanometry after 3 months showed a reduction in mean total resistance from the pretreatment level to 0.77 Pa/cm3/s. We believe that CO2 laser mucotomy is an efficacious, minimally invasive and easy to use treatment of inferior turbinate hypertrophy which is performed under local anesthesia with little discomfort for the patient and does not require hospitalization.

[The influence of endoscopic treatment of isolated, small inflammatory lesions of sphenoid sinus on headache sensation]. / Wplyw leczenia endoskopowego niewielkich, izolowanych zmian zapalnych zatoki klinowej na dolegliwosci bólowe glowy.

UNLABELLED: One of the most frequent complaint in patients with isolated sphenoiditis is headache. In large proportion of these patients no pathologic findings are revealed in sphenoethmoidal recess endoscopicaly thus discrimination between sinus originated and primary headache in such cases may be especially difficult. THE AIM OF THIS STUDY: Was the assessment of the influence of endoscopic treatment of isolated, small inflammatory sphenoid sinus lesions on headache sensation. MATERIAL AND METHOD: 13 patients suffering from headache, with CT-diagnosed isolated, small inflammatory lesions of sphenoid sinus such as mucosal thickening, polypoid tissue and cysts were treated endoscopicaly. Headache was assessed on the basis of five-grade scale before surgery, 1 month after surgery and after the observation period that varied between 5 to 40 months. RESULTS: In the first postoperative month the improvement in their headache was declared by 84.6% of patients, but after longer observation the success rate lowered to 61.5%. The improvement was observed both in patient whose the only complaint was headache and in patients with negative endoscopic findings. CONCLUSIONS: Small isolated sphenoid sinus lesions constitute a group of specific pathologies of paranasal sinuses, which is still difficult to diagnose despite of technological advancement. The treatment of these lesions remains controversial. Decision as to operative treatment should be taken cautiously since headache in great proportion of these patient is not sinus originated. Further multicenter studies on the basis of larger group of patients are necessary to delineate the indications for surgical intervention in patients with small isolated inflammatory sphenoid sinus lesions.

[The pirymethamine influence on hexosoaminidase gene expression in synovial cell culture--preliminary report]. / Wplyw pirymetaminy na ekspresje genów heksozoaminidazy w synowiocytach in vitro--doniesienie wstepne.

UNLABELLED: Inflammation process is leading to increasing of synovial fluid and value of its pressure. Moreover, the impairment of vascular flow within synovial membrane and increased permeability of blood vessels were described. The activity of lysosomal enzymes, such as N-acetyl-beta-glucosaminidase (HEX), was increased in patients with rheumatoid arthritis in comparison to health synovial fluid. It is supposed, that HEX takes part in joint destruction. The using of HEX inhibitors in synovial cell culture and evaluation of HEX mRNA expression before and after the adding of inhibitor may contribute in showing the new ways of understanding pathogenetic pathways of motion organ disorders. THE AIM of the study was to evaluate the expression of HEXA and HEXB genes in the synovial cell culture derived from human synovial inflammatory fluid obtained from patients suffered from rheumatoid arthritis. MATERIAL AND METHODS: The inflamed synovial fluid was taken from patients suffered from rheumatoid arthritis. The following solutions of potential inhibitor--pyrimethamine were used: 20 microg/ml, 10 microg/ml, 3 microg/ml and 1.5 microg/ml. Two separate control groups were established: control group 1 where only 0.6% of ethanol was added to the synovial cell culture; control group 2 where only 0.5% DMSO was added to the synovial cell culture. The relative quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR) was carried out. RESULTS. The difference in HEXA and HEXB expression was observed in synoviocytes obtained in synovial cell culture. Five time higher relative HEXA expression was determined after applying 3 microg/ml of pirymethamine compared with the control 1. The highest concentration of pirymethamine (10 and 20 microg/ml) caused the least elevation of HEXA expression. The slight decreased of HEXB expression was observed under the concentration of pirymethamine: 1.3 and 3 microg/ml. CONCLUSIONS. Pyrimethamine contributes to regulating the HEX gene expression from synovial cells. The change in gene expression level is dependent on the concentration of the pirymethamine. Our preliminary data don't let us establish the concentration of pyrimethamine that may significantly inhibit HEXA and HEXB expression. Further study may be conducted to put new insight into the pathogenesis of joint destruction in the course of rheumatoid arthritis.

[The evaluation of olfactory dysfunction after the operation of ruptured anterior communicating artery aneurysm]. / Ocena zaburzen wechu u chorych po operacji peknietego tetniaka tetnicy laczacej przedniej mózgu.

UNLABELLED: Although the frequency of smell disorders after the operations of the anterior communicating artery aneurysm depends to a large degree on the used surgical approach, several independent of surgery factors may contribute to the postoperative smell tests outcome. THE AIM OF OUR STUDIES: The evaluation of the sense of smell in patients who underwent the operation of ruptured anterior communicating artery aneurysm using pterional approach. MATERIAL AND METHODS: In the retrospective studies the results of smell test of the group of 21 patients operated for ruptured anterior communicating artery aneurysm and 21 healthy volunteers of control group were compared. The patients suffering from neurodegenerative, metabolic and endocrynological disorders, as well as those with rhinoscopic symptoms of rhinosinusitis and with decreased nasal potency confirmed by anterior rhinomanometry were excluded from the study. RESULTS: During the operations it was possible to anatomically retain olfactory nerves in all operated patients. Severe smell disorders (severe hyposmia and anosmia) were detected in 1 (4.7%) patient of the control group and in 6 (28.5%) patients of postoperative group. The mean composite olfactory score in Cain test of the postoperative group was 4.53, and in the control group 5.47 points. The difference did not reach statistic significance (Mann-Whitney test, p = 0.068). CONCLUSIONS: Although pterional approach to anterior communicating artery is relatively low traumatic to olfactory tract, the subarachnoid hemorrhage may have an unfavorable effect on postoperative olfactory nerve functioning. Considering the fact, that in spite of thorough examination of the patients, we were not able to exclude from the studied groups all the patients with preoperatively impaired smell, it seems reasonable to perform similar smell test studies in the prospective way.

Hexosaminidase as a new potential marker for larynx cancer.

OBJECTIVES: Larynx squamous cell carcinoma is one of the most common forms of cancer in the area of the neck. The aim of our study was to investigate the activities of N-acetyl-beta-D-hexosaminidase (HEX) in larynx cancer compared with the specimens from the healthy space of the tumor that served as controls. DESIGN AND METHODS: Larynx cancer (n=15) and normal healthy tissue around the tumor (n=15) were collected from the patients during total laryngectomy. Specimens were immediately frozen in -80 degrees C. To assess hexosaminidase activity, release of p-nitrophenol from p-nitrophenol derivatives was used. RESULTS: We observed a significantly higher activity of the investigated enzyme in all laryngeal cancer specimens compared with that in healthy tissue homogenates. The differences were statistically significant. CONCLUSIONS: It could be assumed that HEX may release particular sugars from the ends of oligosaccharide chains of glycocalyx proteins, changing adhesive forces binding together cells, and the communication between cells and elements of extracellular matrix.

[Possible involvement of hexosaminidase in vocal polyps--preliminary report]. / Rola heksozaminidazy w polipach faldów glosowych--doniesienie wstepne.

UNLABELLED: Vocal fold polyp is usually a result of the injury of the vocal cords. It often appears after vocal overuse or misuse causing trauma in vocal fold mid-membranous and wound formation. THE AIM OF THE STUDY was to assess the activity of hexosaminidase in vocal fold polyps in the comparison to the control tissue. MATERIAL AND METHODS: Vocal polyps (n=8) were collected from 8 patients during direct laryngoscopy. Specimens of normal vocal folds were obtained from 8 cadavers (n=8) served as controls. Specimens were immediately frozen in -80 degrees C. To assess hexosaminidase activity, release of p-nitrophenol from p-nitrophenol derivatives was used. RESULTS: We observed statistical differences between the activity of investigated enzyme in all vocal folds specimens compared with that in normal tissue homogenates. However the activity of HEX achieved a low level. Mean release of HEX from the activated cells in vocal fold specimens was 1.073 nkat/g wet tissue compared with 0.766 nkat/g wet tissue in normal cadaveric vocal fold homogenates. CONCLUSIONS: Low activity of HEX in vocal fold polyps suggests that inflammation may not be a primary factor in the development of the disease and other mechanism should likely be considered in the pathogenesis of vocal fold polyps.

[Hexosoaminidase gene expression in cholesteatoma in adults]. / Ekspresja genów heksozoaminidazy w perlaku ucha srodkowego u doroslych.

BACKGROUND: Middle ear cholesteatoma is a cyst like structure composed of keratinizing squamous epithelium that contains keratin debris and subepithelial connective tissue. Bone resorption may lead to destruction of ossicular chain and temporal bone. The higher activity of N-acetyl-beta3-glucosaminidase (HEX) was noted in cholesteatoma tissue, compared to the controls. It is supposed, that HEX takes part in bone resorption in middle ear cholesteatoma. Using of HEX inhibitors in cultured fibroblasts and evaluation HEX mRNA expression may contribute to showing new ways of understanding cholesteatoma pathogenesis. THE AIM OF THE STUDY: was to elaborate cholesteatoma fibroblast cell culture (CF) technique and evaluate in vitro inhibition potential of pyrimethamine. MATERIAL AND METHODS: Cholesteatoma and normal retroauricular skin sample obtained during surgical treatment were used in the study. CF served as a study group and fibroblast derived from skin specimens--as controls. Pyrimethamine was used at the concentrations of 1.5, 3, 10 and 20 microg/ml. The reverse transcriptase-polymerase chain reaction (RT-PCR) was carried out for the determination of HEX gene expression. RESULTS: RT-PCR established the elevated expression of HEXA and HEXB mRNA in cholesteatoma. HEXA mRNA in CF was six times higher than in the controls. HEX mRNA expression found to be regulated by pyrimethamine. Inhibition of HEXA and HEXB mRNA expression was achieved when the highest concentration of PYR was used. Low concentrations of pyrimethamine upregulated HEX gene in CF. CONCLUSIONS: Pyrimethamine, depending on its concentration, contributes to regulating the HEX gene expression in CF and controls. Pyrimethamine may be regarded as a new future research direction on factors, that may limit development of cholesteatoma.