Vestibular schwannoma surgery causes in majority of patients the unilateral peripheral or combined vestibular lesion due to surgical interruption of both branches of vestibular nerve. It manifests postoperatively with postural instability, vertigo, oscilopsia and even with vegetative symptoms. Central compensation, in which cerebellum plays dominant role, influences duration of the symptoms. Factors affecting compensation may be divided into several groups (general health status of a patient, the tumor itself, actual state of function of vestibular system, external factors induced by a therapeutist). Therapeutic aim is to induce compensatory mechanisms as soon as possible and to reduce overall duration of the compensation. The aim of this study is to consider predictive factors influencing central vestibular compensation and to influence the vestibular compensation itself. Early operated vestibular rehabilitation is a basic procedure. It leads to faster recovery in majority of patients. Even with maximal effort the full compensation is not always achieved. It results in permanent deficit of vestibular function manifested with postural instability. However in some patients it is difficult to achieve full compensation despite maximum effort of a physiotherapeutist and that results in permanent...

The aim of this thesis was to evaluate the effect of rehabilitation with visual biofeedback at acute stage after vestibular schwannoma surgery and to evaluate if preoperative intratympanic aplication of gentamicin has influence on vestibular compensation of standing stability. The study was attended by 20 patients (12 women and 8 men) in the age from 28 to 63 years. Eight patients had vestibular schwannoma diagnosed in their left side, twelve in the right side. Rehabilitation program using interactive rehabilitation system Homebalance took place from the 5th postoperative day until hospital discharge, daily 15 to 20 min. To evaluate of therapeutic intervention objectively the device Synapsys Posturography System was used. Stabilometric examination was done before operation of vestibular schwannoma, after operation and after termination of rehabilitation with visual biofeedback. Statistic analysis showed significant increase of mean values of measured parameters CoP (sway velocity CoP, length of trajectory of CoP, area of the confidence ellipse CoP) postoperatively compared to preoperative values. After termination of therapeutic intervention there was any significant decrease of values of parameters CoP compared to postoperative status. Between group of patients with preoperative aplication of...

The thesis deals with the evaluation of the impact of preoperative application of gentamicin on vestibular compensations of patient after vestibular schwannoma surgery. The trial involved 32 patients with diagnosed vestibular schwannoma and hospitalized with the indicated for neurosurgical resection at the University Hospital Motol. Ototoxic gentamicin was transtympanically applied to ten of these patients two months before their surgery. After the surgery the patients underwent a rehabilitation program based on the visual biofeedback using the Homebalance system. To evaluate the vestibular compensation the subjective visual vertical measurement and the Dynamic Gait Index standardized walking test were used. In total, each patient performed three examinations: before the surgery, after the surgery and before the end of hospitalization. Statistical analysis of the measured data showed the statistically significant increase tilt of subjective visual vertical and the decrease in the overall score of Dynamic Gait Index after surgery. Reduction in the subjective visual vertical tilt and the overall score of Dynamic Gait Index after rehabilitation has not been proven to be statistically significant. Any statistically significant difference was found between the group of patients treated by gentamicin and...

O potencial evocado motor facial (PEMF) tem-se mostrado um excelente método de monitorização do nervo facial, gerando resultados bastante confiáveis e reprodutíveis no que tange à predição da função facial pós-operatória. O critério eletrofisiológico mais utilizado até então para tanto tem sido a relação de amplitude do PEMF final-valor de base. Os objetivos deste trabalho foram avaliar as alterações intraoperatórias da amplitude e da complexidade do PEMF, correlacioná-las com o prognóstico facial no pós-operatório imediato e tardio e verificar se amplitude e complexidade constituem variáveis independentes de predição funcional. Os registros dos potenciais intraoperatórios dos músculos orbiculares do olho e da boca de 35 pacientes portadores de schwannoma vestibular (SV) foram coletados e analisados retrospectivamente de acordo com tempos cirúrgicos preestabelecidos: inicial, abertura da dura-máter, dissecação do tumor (TuDis), ressecção do tumor (TuRes) e final. No pós-operatório imediato, a função facial apresentou uma significativa correlação negativa com as relações de amplitude do PEMF durante a TuDis, a TuRes e ao final do procedimento nos músculos orbiculares do olho (p =0,003, 0,055 e 0,028, respectivamente) e da boca (p=0,002, 0,104 e 0,014, respectivamente). No último seguimento, entretanto, a correlação foi significativa apenas para o músculo orbicular da boca, durante a TuDis (p=0,005) e ao final do procedimento (p=0,102). As variações da complexidade dos potenciais alcançaram resultados mais significativos tanto no pós-operatório imediato, quanto no tardio, de forma que houve uma correlação negativa no músculo orbicular do olho apenas nas medidas finais (imediato, p=0;023; seguimento, p=0,116) e no músculo orbicular da boca durante a TuDis, a TuRes e a medida final (imediato, p=0,071, 0,000 e 0,001, respectivamente; seguimento, p=0,015, 0,001 e 0,01, respectivamente). As alterações intraoperatórias das relações de amplitude e de complexidade dos PEMFs parecem representar variáveis independentes, podendo ser utilizadas na predição da função facial pós-operatória durante cirurgias de ressecção de SV. Baseados nos resultados deste trabalho, a monitorização evento-valor de base é bastante útil, justificando mudanças imediatas da estratégia cirúrgica, com o intuito de reduzir as chances de uma lesão definitiva do nervo facial. / Facial motor evoked potential (FMEP) amplitude ratio reduction at the end of the surgery has been identified as a good predictor for postoperative facial nerve outcome. We sought to investigate variations in FMEP amplitude and waveform morphology during vestibular schwannoma (VS) resection and to correlate these measures with postoperative facial function immediately after surgery and at the last follow-up. Besides we analyzed the relationship between quantitative parameters. Intraoperative orbicularis oculi and oris muscles FMEP data from 35 patients undergoing surgery for VS resection were collected, then analyzed by surgical stage: initial, dural opening, tumor dissection (TuDis), tumor resection (TuRes) and final. Immediately after surgery, postoperative facial function correlated significantly with the FMEP amplitude ratio during TuDis, TuRes and final in both the orbicularis oculi (p´s=0.003, 0.055 and 0.028, respectively) and oris muscles (p´s=0.002, 0.104 and 0.014, respectively). At the last follow-up, however, facial function correlated significantly with the FMEP amplitude ratio only during TuDis (p=0.005) and final (p=0.102) for the orbicularis oris muscle. At both time points, postoperative facial paresis correlated significantly with FMEP waveform deterioration in orbicularis oculi during final (immediate, p=0.023; follow-up, p=0.116) and in orbicularis oris during TuDis, TuRes and final (immediate, p´s=0.071, 0.000 and 0.001, respectively; follow-up, p´s=0.015, 0.001 and 0.01, respectively). FMEP amplitude ratio and waveform morphology during VS resection seem to represent independent quantitative parameters that can be used to predict postoperative facial function. Event-to-baseline FMEP monitoring is quite useful to dictate when intraoperative changes in surgical strategy are warranted to reduce chances of facial nerve injury.

<p>Neurofibromatosis type-2 (NF2) is an autosomal dominant disorder with the clinical hallmark of bilateral eighth cranial nerve schwannomas. However, the diagnostic criterion is complicated by the presence of a variable phenotype, with the severe form presenting with additional tumors such as peripheral schwannoma, meningioma and ependymoma. We constructed a microarray spanning 11Mb of 22q, encompassing the <i>NF2 </i>gene, to detect deletions in schwannoma. Forty seven patients were analyzed and heterozygous deletions were detected in 45% of tumors. Using this array-based approach, we also detected genetic heterogeneity in a number of samples studied. Despite the high sensitivity and the comprehensive series of studied schwannomas, no homozygous deletions affecting the <i>NF2</i> gene were detected <b>(paper I)</b>. In order to detect more subtle deletions within the <i>NF2</i> locus, a higher-resolution gene-specific array was developed, for the detection of disease-causing<b> </b>deletions using a PCR-based non-redundant strategy. This novel approach for array construction significantly increased the reliability and resolution of deletion-detection within the <i>NF2 </i>locus <b>(paper II)</b>. To further expand the coverage of the 11 Mb microarray, we constructed the first comprehensive microarray representing a human chromosome for analysis of DNA copy number. This 22q array covers 34.7 Mb, representing 1.1% of the genome, with an average resolution of 75 kb <b>(paper III)</b>. Using this array, we analyzed sporadic and familial schwannomatosis samples, which revealed two commonly deleted regions within the immunoglobulin lambda locus and the <i>GSTT1/CABIN1</i> locus. These regions were further characterized using higher-resolution non-redundant arrays, bioinformatic tools, positional cloning and mutational screening. Missense mutations were detected in the <i>CABIN1</i> gene, which may contribute to the pathogenesis of schwannomatosis and therefore requires further study <b>(paper IV)</b>. Meningioma is the second most common NF2-associated tumor and loss of 1p has been previously established as a major genetic factor for disease initiation/progression and also correlates with increased morbidity. We analyzed 82 meningiomas using a chromosome 1 tiling-path genomic microarray. The distribution of aberrations detected supports the existence of at least four regions on chromosome 1, which are important for meningioma tumorigenesis <b>(paper V)</b>.</p>

<p>The array-based form of comparative genomic hybridization (array-CGH) is a new methodology that has shown to be of significant importance. This thesis focuses on the development of array-CGH with the aim to define candidate regions/genes on chromosome 22 in a wide spectrum of cancer-related conditions. In <b>paper I</b>, we developed and applied the first comprehensive genomic microarray, representing human chromosome 22, for analysis of DNA copy number. Using this array-based approach, we identified gene copy number alterations, including heterozygous/homozygous deletions, amplifications, IGLV/IGLC locus instability and the breakpoints of imbalanced translocation, in several 22q-associated disorders. In <b>paper II</b>, we applied the same array to perform DNA copy number profiling of a series of ovarian carcinoma. cDNA arrays were also used in this study to correlate gene expression levels with DNA-copy number. In the course of this analysis, we determined a small 3.5 Mb candidate 22q telomeric region and suggested a number of specific candidate genes. <b>Paper III</b> described the comprehensive and high-resolution analysis of chromosome 22 in a large set of various stage breast cancers. Multiple distinct patterns of genetic aberrations were observed. The smallest identified candidate locus was 220 kb in size and mapped to a gene-rich region in the vicinity of telomere of 22q. Intriguing result of this study was the detection of high frequency (26.6%) of intra-tumoral clonal variation in gene copy number profiles, which should be viewed as a high number, considering that we study in detail only a single human chromosome. In <b>paper IV</b>, we profiled a series of 28 Wilms tumor samples using 22q-array in order to assess specific regions affected with DNA dosage-alterations. The distribution of aberrations defined a complex amplifier genotype and delimited two tumor suppressor/oncogene candidate loci. These results open up for several avenues for continued research of these tumor forms. These findings also demonstrate the power of array-CGH in the precise determination of minute DNA copy number alterations and strengthen the notion that further studies, preferentially in the context of the entire human genome, are needed.</p>

<p>Neurofibromatosis type 2 (NF2) is an autosomal dominant disorder with the clinical hallmark of bilateral vestibular schwannomas (VS). Patients affected by a severe NF2 phenotype also presents with peripheral schwannomas, meningiomas and ependymomas. The closely related disorder schwannomatosis also displays multiple schwannomas, but never VS. Mutation screening of the <i>NF2</i> gene in the above mentioned tumors did not identify mutations in numerous of cases. We analyzed the DNA sequence covering the <i>NF2</i> locus in order to identify evolutionarily conserved non-genic sequences (CNGs) with unknown regulatory function (paper I). The aim was to analyze CNGs for mutations in DNA derived from patients affected by NF2 associated tumors. During mutation analysis of the coding part of <i>NF2</i> and within the CNGs defined in paper I, were mutations detected in 39% of sporadic meningiomas (paper II). Two candidate regions were identified on 22q using array-CGH. Methylation profiling did not identify methylation of the <i>NF2</i> promoter in these tumors. Sporadic schwannomas were profiled for CNV using a 22q genomic array in the search for putative gene(s) that in addition to <i>NF2</i> could be involved in the development of schwannoma and/or schwannomatosis (paper III). The predominant aberration identified was monosomy 22. Terminal and interstitial deletions encompassing the <i>NF2</i> gene were detected in tumor DNA and eight loci affected by CNV in constitutional DNA. Some of these CNVs are unlikely to be phenotypically neutral, considering their size and gene content. Two schwannomatosis candidate regions were identified on 22q using array-CGH (paper IV). These regions were further characterized by a PCR-product based array with higher resolution. Rearrangements of the immunoglobulin lambda (<i>IGL</i>) locus detected were restricted to schwannomatosis patients. In the second candidate region spanning <i>GSTT1</i> and <i>CABIN1</i> genes, was frequent copy number polymorphism at the <i>GSTT1</i> locus identified. We further describe missense mutations in the <i>CABIN1 </i>gene, making this gene a plausible candidate which may contribute to the pathogenesis of these disorders. </p>

Neurofibromatosis type-2 (NF2) is an autosomal dominant disorder with the clinical hallmark of bilateral eighth cranial nerve schwannomas. However, the diagnostic criterion is complicated by the presence of a variable phenotype, with the severe form presenting with additional tumors such as peripheral schwannoma, meningioma and ependymoma. We constructed a microarray spanning 11Mb of 22q, encompassing the NF2 gene, to detect deletions in schwannoma. Forty seven patients were analyzed and heterozygous deletions were detected in 45% of tumors. Using this array-based approach, we also detected genetic heterogeneity in a number of samples studied. Despite the high sensitivity and the comprehensive series of studied schwannomas, no homozygous deletions affecting the NF2 gene were detected <b>(paper I)</b>. In order to detect more subtle deletions within the NF2 locus, a higher-resolution gene-specific array was developed, for the detection of disease-causing<b> </b>deletions using a PCR-based non-redundant strategy. This novel approach for array construction significantly increased the reliability and resolution of deletion-detection within the NF2 locus <b>(paper II)</b>. To further expand the coverage of the 11 Mb microarray, we constructed the first comprehensive microarray representing a human chromosome for analysis of DNA copy number. This 22q array covers 34.7 Mb, representing 1.1% of the genome, with an average resolution of 75 kb <b>(paper III)</b>. Using this array, we analyzed sporadic and familial schwannomatosis samples, which revealed two commonly deleted regions within the immunoglobulin lambda locus and the GSTT1/CABIN1 locus. These regions were further characterized using higher-resolution non-redundant arrays, bioinformatic tools, positional cloning and mutational screening. Missense mutations were detected in the CABIN1 gene, which may contribute to the pathogenesis of schwannomatosis and therefore requires further study <b>(paper IV)</b>. Meningioma is the second most common NF2-associated tumor and loss of 1p has been previously established as a major genetic factor for disease initiation/progression and also correlates with increased morbidity. We analyzed 82 meningiomas using a chromosome 1 tiling-path genomic microarray. The distribution of aberrations detected supports the existence of at least four regions on chromosome 1, which are important for meningioma tumorigenesis <b>(paper V)</b>.

The array-based form of comparative genomic hybridization (array-CGH) is a new methodology that has shown to be of significant importance. This thesis focuses on the development of array-CGH with the aim to define candidate regions/genes on chromosome 22 in a wide spectrum of cancer-related conditions. In <b>paper I</b>, we developed and applied the first comprehensive genomic microarray, representing human chromosome 22, for analysis of DNA copy number. Using this array-based approach, we identified gene copy number alterations, including heterozygous/homozygous deletions, amplifications, IGLV/IGLC locus instability and the breakpoints of imbalanced translocation, in several 22q-associated disorders. In <b>paper II</b>, we applied the same array to perform DNA copy number profiling of a series of ovarian carcinoma. cDNA arrays were also used in this study to correlate gene expression levels with DNA-copy number. In the course of this analysis, we determined a small 3.5 Mb candidate 22q telomeric region and suggested a number of specific candidate genes. <b>Paper III</b> described the comprehensive and high-resolution analysis of chromosome 22 in a large set of various stage breast cancers. Multiple distinct patterns of genetic aberrations were observed. The smallest identified candidate locus was 220 kb in size and mapped to a gene-rich region in the vicinity of telomere of 22q. Intriguing result of this study was the detection of high frequency (26.6%) of intra-tumoral clonal variation in gene copy number profiles, which should be viewed as a high number, considering that we study in detail only a single human chromosome. In <b>paper IV</b>, we profiled a series of 28 Wilms tumor samples using 22q-array in order to assess specific regions affected with DNA dosage-alterations. The distribution of aberrations defined a complex amplifier genotype and delimited two tumor suppressor/oncogene candidate loci. These results open up for several avenues for continued research of these tumor forms. These findings also demonstrate the power of array-CGH in the precise determination of minute DNA copy number alterations and strengthen the notion that further studies, preferentially in the context of the entire human genome, are needed.

Neurofibromatosis type 2 (NF2) is an autosomal dominant disorder with the clinical hallmark of bilateral vestibular schwannomas (VS). Patients affected by a severe NF2 phenotype also presents with peripheral schwannomas, meningiomas and ependymomas. The closely related disorder schwannomatosis also displays multiple schwannomas, but never VS. Mutation screening of the NF2 gene in the above mentioned tumors did not identify mutations in numerous of cases. We analyzed the DNA sequence covering the NF2 locus in order to identify evolutionarily conserved non-genic sequences (CNGs) with unknown regulatory function (paper I). The aim was to analyze CNGs for mutations in DNA derived from patients affected by NF2 associated tumors. During mutation analysis of the coding part of NF2 and within the CNGs defined in paper I, were mutations detected in 39% of sporadic meningiomas (paper II). Two candidate regions were identified on 22q using array-CGH. Methylation profiling did not identify methylation of the NF2 promoter in these tumors. Sporadic schwannomas were profiled for CNV using a 22q genomic array in the search for putative gene(s) that in addition to NF2 could be involved in the development of schwannoma and/or schwannomatosis (paper III). The predominant aberration identified was monosomy 22. Terminal and interstitial deletions encompassing the NF2 gene were detected in tumor DNA and eight loci affected by CNV in constitutional DNA. Some of these CNVs are unlikely to be phenotypically neutral, considering their size and gene content. Two schwannomatosis candidate regions were identified on 22q using array-CGH (paper IV). These regions were further characterized by a PCR-product based array with higher resolution. Rearrangements of the immunoglobulin lambda (IGL) locus detected were restricted to schwannomatosis patients. In the second candidate region spanning GSTT1 and CABIN1 genes, was frequent copy number polymorphism at the GSTT1 locus identified. We further describe missense mutations in the CABIN1 gene, making this gene a plausible candidate which may contribute to the pathogenesis of these disorders.