Comprehensive high-resolution genomic profiling and cytogenetics of human chondrocyte cultures by GTG-banding, locus-specific FISH, SKY and SNP array.

In the development of cell-based medicinal products, it is crucial to guarantee that the application of such an advanced therapy medicinal product (ATMP) is safe for the patients. The consensus of the European regulatory authorities is: "In conclusion, on the basis of the state of art, conventional karyotyping can be considered a valuable and useful technique to analyse chromosomal stability during preclinical studies". 408 chondrocyte samples (84 monolayers and 324 spheroids) from six patients were analysed using trypsin-Giemsa staining, spectral karyotyping and fluorescence in situ hybridisation, to evaluate the genetic stability of chondrocyte samples from non-clinical studies. Single nucleotide polymorphism (SNP) array analysis was performed on chondrocyte spheroids from five of the six donors. Applying this combination of techniques, the genetic analyses performed revealed no significant genetic instability until passage 3 in monolayer cells and interphase cells from spheroid cultures at different time points. Clonal occurrence of polyploid metaphases and endoreduplications were identified associated with prolonged cultivation time. Also, gonosomal losses were observed in chondrocyte spheroids, with increasing passage and duration of the differentiation phase. Interestingly, in one of the donors, chromosomal aberrations that are also described in extraskeletal myxoid chondrosarcoma were identified. The SNP array analysis exhibited chromosomal aberrations in two donors and copy neutral losses of heterozygosity regions in four donors. This study showed the necessity of combined genetic analyses at defined cultivation time points in quality studies within the field of cell therapy.

The use of 5-aminolevulinic acid fluorescence guidance in resection of pediatric brain tumors.

INTRODUCTION: Whereas in the adult population 5-Aminolevulinic acid (5-ALA) fluorescence guidance has been widely accepted for improving the extent of tumor resection, the application in children remains an off-label use. Even though most pediatric study protocols require a complete resection for improving outcome parameters, only few pediatric patients have been operated with fluorescence guidance, and it remains questionable, whether and which pediatric tumors show useful fluorescence. We present casuistic reports of application of 5-ALA in children collected from three different neurosurgical departments. PATIENTS AND METHODS: In children with suspected malignant intracerebral tumor or recurrence, individual informed consent was obtained in each case from the parents. 5-ALA was administered according to the adult protocol, with 20 mg/kg, 2 h before induction of anesthesia. We retrospectively analyzed 18 patients (13 male, 5 female; age 3-18 years), using the intraoperative neurosurgical protocol, the postoperative MRI results, and the follow-up clinical examinations. RESULTS: The use of 5-ALA fluorescence guidance proved to be safe in our group of pediatric patients. Fluorescence guidance was most useful for recurrent glioblastoma resection. Medulloblastoma tissue displayed fluorescence only inconsistently, and most pilocytic astrocytoma remained without staining. Ganglioglioma showed partial staining in the central tumor areas, without allowing the use for circumferent resection. CONCLUSION: The off-label use of 5-ALA fluorescence guidance in pediatric patients appears to be most useful in recurrent high-grade gliomas. Fluorescence accumulation in other pediatric brain tumor entities is not predictable and should be evaluated in future clinical studies before being integrated into the current treatment protocols.

Characterization of chromosomal aberrations in a case of glioblastoma multiforme combining cytogenetic and molecular cytogenetic techniques.

A case of glioblastoma multiforme (GBM) that was investigated with a broad spectrum of cytogenetic and molecular cytogenetic techniques is reported. The results of cytogenetic studies, interphase fluorescence in situ hybridization, comparative genomic hybridization, and spectral karyotyping (SKY) are reported. Various structural chromosomal aberrations were identified, among which aberrations involving chromosome arm 2p were especially frequent. Using SKY, six translocations not previously described in GBM are reported.

Management results attained by predominantly late surgery for intracranial aneurysms.

In contrast to previous studies conducted by various authors, who recommended early surgery for all patients admitted to the hospital within 72 hours of an aneurysmal subarachnoid hemorrhage, several more recent studies have declined to advise early surgery for the treatment of patients with impaired consciousness. In our series, early surgery was undertaken for patients who were rated at Grades 1 to 2 (Hunt and Hess) at admission and who did not exhibit any additional risk factors (e.g., evidence of incipient vasospasm, giant aneurysm, unfavorable aneurysm location, or a severe concomitant disease). Only three patients rated Grade 3 at admission with a favorable aneurysm location and shape underwent early surgery. The management results attained in this series (n = 131), in which the early surgery rate was 17%, have been analyzed. The management mortality rate of patients with aneurysmal subarachnoid hemorrhage was 13%, and it was 7.7% for patients admitted at Grades 1 to 3 on the Hunt and Hess scale. Good results (Glasgow Outcome Scale, 1 or 2) were attained in 75% of the entire study population, in 85% of patients admitted at Grades 1 to 3, and in 53% of those patients who were admitted at Grades 4 to 5 and who underwent late surgery after their condition had improved to Grades 1 to 3. At an average interval of 3 years after the operation, 83% of the patients discharged with Glasgow Outcome Scale ratings of 1 or 2 reported no significant restriction of their "stress resistance."(ABSTRACT TRUNCATED AT 250 WORDS)

Intradiploic arachnoid cyst with extensive deformation of craniofacial osseous structures: case report.

OBJECTIVE AND IMPORTANCE: A unique case of a large intradiploic arachnoid cyst involving craniofacial osseous structures is reported. CLINICAL PRESENTATION: The patient presented with a hard mass in the right frontal region, proptosis, and inferior globe displacement. Computed tomography revealed an intraosseous cyst of cerebrospinal fluid intensity with extension from the anterior cranial fossa to the infratemporal fossa. INTERVENTION: After resection of the cyst wall and closure of two small round dural defects, the involved craniofacial region was reconstructed. CONCLUSION: The medical history of the patient and the intraoperative observations support the contention that the cyst in the reported case was congenital in origin. The features concerned with diagnosis and pathogenesis of this rare entity are discussed.

The expression of angiogenin in tissue samples of different brain tumours and cultured glioma cells.

BACKGROUND: As a potential angiogenetic factor the 14.1 kDa polypeptide angiogenin induces neovascularisation. MATERIALS AND METHODS: We investigated the angiogenin expression by immunoblotting and an ELISA in 60 tissue specimens (40 gliomas, 20 other intracranial tumours), in 22 glioma cell cultures and in 4 supernantants of cultivated glioblastoma cells. RESULTS: We could show that angiogenin is detectable in different kinds of intracranial tumours with the highest amount in meningiomas and the lowest amount in low grade astrocytomas. In tissue specimens, a significantly higher angiogenin expression was measured in meningiomas compared to gliomas and metastases. Angiogenin could be detected in primary cultivated glioma cells, but not in the permanent cell lines. There was a significant correlation to the malignancy within the gliomas with an increase of angiogenin concentration according to the higher grade of malignancy. CONCLUSIONS: Our data suggest that angiogenin may contribute to the malignant transformation of gliomas and could perhaps advise that the physiological role of angiogenin is not restricted exclusively to angiogenesis. Based on these findings the clinical importance of angiogenin for therapeutic decisions in malignant brain tumours remains unclear and further analyses on m-RNA-levels are required.

Cerebrospinal fluid leak after microsurgical surgery in vestibular schwannomas via retrosigmoidal craniotomy.

OBJECTIVE: Cerebrospinal fluid (CSF) leak is still a common complication in surgery of vestibular schwannoma, increasing morbidity and prolonging hospital stay. Our single center study was performed to determine the incidences of CSF leaks after microsurgical removal of vestibular schwannoma via a retrosigmoidal approach with two different surgical closure techniques. METHODS: Between January 2003 and December 2009 in 81 patients, microsurgical tumor resection using a suboccipital, retrosigmoidal approach was performed with an interdisciplinary ENT and neurosurgical management was performed. In 41 cases, the dural closure was done using a sandwich technique: subdural closure with TissuFleece® respectively Spongostan®, and after that dural suture and epidural Tachosil® were fixed on. In 40 cases, the dura was sealed epidurally with Tachosil after suture. In 65 cases, the posterior wall of the petrous bone was drilled. The closure was performed using muscle and FibrinGlue®. All patients had a minimal follow-up of 1 year. RESULTS: Seven patients (8.6%) developed a CSF fistula. Three patients (3.7%) underwent surgical procedure because of persisting CSF fistula while in four cases (4.9%) spontaneous closure under lumbar drain was observed. Comparing the different techniques of dural sealing, we found in 41 patients with sandwich technique three CSF leaks (7.3%) while there were four CSF leaks (10%) in 40 patients with a single epidurally sealed dural closure (P=0.69). No rhinorrhea or otorhinorrhea was observed. No intracranial infection or meningitis in case of CSF leak occurred. CONCLUSION: Suture and occlusion of the dura is an important step to prevent CSF leak and postoperative infection. By comparing sandwich technique and single-layer dural sealing, no significant difference could be shown.

Clinical results of the foraminotomy as described by Frykholm for the treatment of lateral cervical disc herniation.

During the period between 1980 and 1989, foraminotomy as described by Frykholm was performed on 230 patients suffering from lateral cervical disc herniation with exclusively radicular symptomatology. Following an average postoperative period of 3.5 years, the subjective alleviation of symptoms and the neurological symptomatology were examined in 161 patients. The mortality was nil; the morbidity was 5%, with a rapid full recovery in 4% of cases. In addition to the intra-operative findings of soft and hard disc lesions, a third group with combined findings was created. Excellent or good results were obtained in 98% of the patients with soft disc lesions, in 91% of the patients with combined findings, and in 84% of those with hard disc lesions. In 93% of the cases, there was a complete or marked improvement of paresis; in 82%, of the sensory deficits. A total of 92% of the patients were able to carry out their previous occupation to the full extent. The high efficacy of foraminotomy and the low incidence of complications described in previous studies was thereby confirmed.

Treatment of chronic subdural haematoma with burr-hole craniostomy and closed drainage.

Despite the high incidence of chronic subdural haematoma (SDH), to date relatively few authors have undertaken a systematic analysis of the results attained following burr-hole craniostomy and closed-system drainage on the basis of a study involving a large patient population defined according to clear-cut criteria. Between 1980 and 1993, surgery was performed on 212 patients with a chronic SDH. In a retrospective study, the condition of each patient was assessed at the time of admission on the basis of the Bender Scale and the results 4 weeks after discharge were classified according to the Glasgow Outcome Scale (GOS). At admission, 165 patients (78%) were in a satisfactory condition (Stage 1 or 2); 47 (22%) were at Stage 3 or 4, i.e. stuporous or comatose. Nine (4%) of the patients died; none of these deaths was attributable to the operation. In addition to significant extracerebral disease (2.4%), CT revealed ischaemic cerebral infarctions in three patients (1.4%) after removal of the SDH. The operative morbidity was 4.2%. In 22% of the patients, surgical intervention was required to remove a recurring SDH. In 90% of the patients, the results achieved could be graded as 'very good' or 'good' (GOS 1 or 2). The results were related to the condition at admission (p < 0.001) and the patient's age (p < 0.05). Factors that did not have a significant effect on the outcome included whether the SDH formation was bilateral or unilateral, the extent of neomembranous organization of the haematoma, and the amount of primary cerebral expansion following decompression. We were able to confirm the results of previous studies that the method used is a highly effective treatment, with a low incidence of complications.

Informed consent in neurosurgery: patients' recall of preoperative discussion.

OBJECTIVE: Informed consent (IC) is an important principle of modern medicine and the quality of the process is likely to receive increasing attention in future due to complex surgical procedures and a development of social mistrust for medical treatment. Medico-legal action is also becoming an important influence on IC, in particular the extent of warning to be given about the degree of risk. Evaluation of IC, however, encounters various problems. One key element of a knowledgeable decision is an analysis based on the disclosed risks. METHODS: In a prospective study, 104 consecutive patients were asked to write down as many items of the list of risks as possible two hours after consent interview. The sample included adults (mean age: 52 years) without any significant neuropsychological impairment who underwent elective intracranial (52%) or spinal (48%) surgery. Consent interview based on the proposals of the Deutsche Gesellschaft für Neurochirurgie and the Berufsverband Deutscher Neurochirurgen 1998. The average number of disclosed risks was 32 in intracranial and 25 in spinal surgery. According to the results in the literature 6 'typical major risks' of the proposed treatment were selected. RESULTS: The median value of the total of recalled risks was 4 in the spinal group and 5 in the intracranial group. The mean score of general information retention was 18%. 65% of the patients did not recall more than 2 of 6 'typical major risks'. No significant correlation between recall performance and age or education was found. CONCLUSION: The general principles of memory apply and have implications in states of emotional arousal. Whereas education and age, unlike in previous reports, did not appear to influence recall, thus indicating that we had succeeded in tailoring IC to the individual, recall rate was low in most cases. Physicians should highlight the discrepancy and conflict between the requirements for the defence of medical practice in the law courts and the actual interests of patients. They should promote research to establish what really is 'appropriately informed consent'.