Meningitis and spondylodiscitis due to Nocardia nova in an immunocompetent patient.

BACKGROUND: Disseminated nocardiosis is a very rare disease. By now only few cases of meningitis and spondylodiscitis have been reported. To our knowledge, this is the first case of meningitis caused by Nocardia nova. CASE PRESENTATION: We report on a case of bacteraemia, meningitis and spondylodiscitis caused by N. nova in an immunocompetent patient. We describe the long, difficult path to diagnosis, which took two months, including all diagnostic pitfalls. After nocardiosis was diagnosed, intravenous antibiotic therapy with ceftriaxone, later switched to imipenem/cilastatin and amikacin, led to rapid clinical improvement. Intravenous therapy was followed by oral consolidation with co-trimoxazole for 9 months without any relapse within 4 years. CONCLUSIONS: Establishing a diagnosis of nocardiosis is a precondition for successful antibiotic therapy. This requires close communication between clinicians and laboratory staff about the suspicion of nocardiosis, than leading to prolonged cultures and specific laboratory methods, e.g. identification by 16S rDNA PCR.

Is there any use? Validity of 4D rasterstereography compared to EOS 3D X-ray imaging in patients with degenerative disk disease.

INTRODUCTION: Previous studies of 4D rasterstereography show a high intra- and interday reliability. However, only few studies validate rasterstereography to conventional X-ray imaging. We utilized EOS X-ray imaging system (EOS Imaging, Paris, France) for accurate 3D spinal modeling and compared the results to parameters obtained by 4D rasterstereography. The aim of the present study was to validate 4D rasterstereography in patients with degenerative disk disease (DDD). MATERIALS AND METHOD: Thirty-four individuals with DDD (female = 22 and male = 12) were included. EOS X-ray images were analyzed to determine spinal [lumbar lordosis (LL) and thoracic kyphosis (TK)] and pelvic parameters [pelvic obliquity (PO) and pelvic axial rotation (PR)]. Patients received 4D rasterstereographic measurements on the same day as EOS imaging. Parameters obtained by rasterstereography were compared to those obtained by EOS X-ray imaging. We used Bland and Altman's test as well as Pearson test to validate rasterstereography. Additionally, we calculated interrater reliability of EOS X-ray analysis using the intraclass correlation coefficient (ICC). RESULTS: Our data showed only weak correlation between 4D rasterstereography and EOS X-ray imaging for spinal parameters (LL and TK). Pelvic parameters (PO and PR) showed no correlation. Interrater correlation reliability for EOS analysis was excellent (ICC > 0.8). CONCLUSION: Our data suggest that rasterstereographic systems are no reliable substitute for X-ray-based imaging systems in patients with degenerative disk disease. EOS imaging was shown to provide reliable and accurate spinal modeling. Based on our results, rasterstereographic imaging should be used with caution for evaluating spinal and pelvic parameters in patients with DDD. These slides can be retrieved under Electronic Supplementary Material.

Skeletal spread of an anaplastic astrocytoma (WHO grade III) and preservation of histopathological properties within metastases.

BACKGROUND: The incidence of extraneural metastases of glioma is low. Metastases occur at different sites and, infrequently, as diffuse bone marrow infiltration. Direct contact of a glioma with extrameningeal tissues might be a reason for extraneural metastases. However, the role of haematogenous spread remains unclear. METHODS: We report on a young patient who suffered from a left frontal anaplastic WHO grade III astrocytoma, which was treated with gross total resection and irradiation (60 Gy). No local relapse occurred during the following course, but a diffuse infiltration of the bone marrow was diagnosed 12 months after the initial diagnosis. The patient died 6 months later, as a result of hypercalcaemia and pancytopenia. The histopathological properties of the tumour and its bone metastases were analysed, as well as the mutations of the isocitrate dehydrogenase 1 gene (IDH1). To study the route of tumour dissemination, the peripheral blood of the patient was analysed for circulating tumour cells (CTCs). RESULTS: This study describes a rare case of an extraneurally metastasised WHO grade III anaplastic astrocytoma. The occurrence of bone marrow infiltration coinciding with the finding of a stable intracranial tumour is a notably unusual situation. The properties of the primary tumour were maintained within the metastases in our patient. No CTCs were found in the peripheral blood at one random time point after the diagnosis of bone metastases. CONCLUSIONS: Despite young patient age, a stable intracranial course with a single location and mutations in the IDH1 gene, the patient's overall survival was short at 18 months after diagnosis. This finding illustrates the therapeutic dilemma in patients with bone marrow involvement complicating the use of alkylating agents, such as temozolomide. Repeated and systematic blood sampling in a large cohort of patients is needed for the detection of CTCs in glioma patients with systemic tumour spread. Future studies investigating how intrinsic factors in glioma cell biology cause rare metastases in these tumours are needed.

A comprehensive analysis of vascular complications in 3,889 glioma patients from the German Glioma Network.

Ischemic strokes, intracranial hemorrhages (ICH) and deep venous thromboembolism (DVT) are clinically important events in patients with gliomas. In this multicentre, noninterventional observational study, current data pertaining to frequency, contributing factors and outcomes of vascular events during times of anti-angiogenic therapy with the antibody against vascular endothelial growth factor, bevacizumab (BEV) was collected from the German Glioma Network. Among 3,889 glioma patients, 70 ischemic strokes (1.8 %) and 123 ICH (3.2 %) were recorded. 143 DVT (5.0 %) were recorded in 2,855 patients. Rates of DVT and ICH, but not of ischemic strokes, increased with the World Health Organization (WHO) grade of glioma. In 81 BEV-treated patients, five ischemic strokes (6.2 %), one ICH (1.2 %) and six DVT (7.4 %) were documented. Compared to patients that were not treated with BEV, ischemic stroke rate was significantly higher during treatment with BEV (p < 0.001). The rates of DVT (p = 0.123) or ICH (p = 0.571) in BEV-treated patients did not differ. On cerebral magnetic resonance imaging (MRI), BEV-related ischemic strokes appeared as diffusion-restricted sites next to contrast-enhancing tumor. 67 % of ICH, 61 % of ischemic strokes and 18 % of DVT occurred postoperatively (within 30 days after tumor resection). Outcome after postoperative ICH was significantly worse than after spontaneous ICH (p = 0.008). Ischemic stroke outcomes did not differ between postoperative and spontaneous occurrence (p = 0.401). Rate of pulmonary embolism did not differ significantly between postoperative and spontaneous DVT (p = 0.133). Relatively low rates of ICH and DVT might be partially due to a high proportion of low-grade gliomas in this patient cohort. The finding of a relevant number of symptomatic, therapy-associated intracerebral diffusion restrictions should be controlled in ongoing phase III studies.

Correlation between the severity of neurological deficits in patients with symptomatic spinal metastases and short-term postoperative functional neurological improvements.

OBJECTIVE: The correlation between the severity of neurological deficit of patients with symptomatic spinal metastases and short-term, postoperative, functional improvements after surgery is unclear. The aim of this clinical trial was to determine the influence of neurological deficit severity on short-term functional outcomes after surgery, and to devise optimal treatment strategies for this patient population. METHODS: Between 01/1999 and 12/2004, 194 patients with symptomatic spinal metastases were surgically treated. Each patient underwent neurological examination pre- and postoperatively. The results were ranked according to the Frankel score. The pre- and postoperative Frankel score was compared in order to assess the development of neurostatus following a surgical procedure. RESULTS: Complete postoperative remission was seen in 27% of all patients (27/101) with a preoperative Frankel score D, as well as in 5% of all patients (4/77) with a preoperative Frankel score C. Improvement of the preoperative neurologic deficits occurred in 27% of all patients (27/101) with a preoperative Frankel score D, 58% of all patients (45/77) with a preoperative Frankel score C, 36% of all patients (5/14) with a preoperative Frankel score B, and 50% of all patients (1/2) with a preoperative Frankel score A. CONCLUSION: Surgical treatment of patients suffering from neurological deficits caused by spinal metastases results in neurological function improvements. The probability of neurological improvement does not correlate with the severity of the preoperative neurologic deficit. As a consequence, the decision to operate on these patients should not depend on the severity of pre-operative neurological symptoms alone.

A survey on clinical study participation and study infrastructure at non-university hospitals in Germany.

OBJECTIVE: In order to participate in multicenter clinical trials a fair amount of infrastructure and human resources has to be provided by hospitals. Therefore clinical trials are carried out predominantly in university hospitals. Data concerning participation in clinical trials and the infrastructure of study centers in non-university hospitals in Germany do not exist. A survey was thus conducted to evaluate the current status of clinical study performance in non-university hospitals. MATERIALS AND METHODS: A questionnaire comprising 10 questions covering hospital infrastructure, local study history, and the individual interest in performing studies was sent to 790 non-university hospitals in Germany. RESULTS: 58.7% of questionnaires were returned for evaluation. 74.1% of nonuniversity hospitals participate in clinical studies. Hospital size is a significant predictor of study participation. 25.5% of hospitals have established a multidisciplinary study center. 86.2% have a certified study nurse and in 34.5% this nurse is the only person running the study center. Only 25.5% of hospitals were not interested in participating in clinical studies at all, even if an individual tailored concept were to be offered. CONCLUSIONS: The demand for more hospitals to participate in clinical trials is urgent since high quality studies are a fundamental part of clinical research. Even though 75% of non-university hospitals in Germany already participate in clinical trials, it may be possible to increase this number. In addition by establishing and developing study centers in hospitals the quality of studies will presumably rise, and due to the concentration of study resources, the number of clinical trials may increase.

Non-conventional treatments sought by patients after surgery for intramedullary spinal cord tumors.

OBJECTIVE: Despite modern microsurgical techniques and interdisciplinary treatment options, intramedullary spinal cord tumors often lead to profound neurological deficits. Some patients may search for non-scientific or unconventional therapeutic options to treat the symptoms induced by the tumor itself or arising from treatment. The extent of non-scientific therapy use, the rationale behind it, and costs of these forms of therapy are unknown. METHODS: A questionnaire consisting of 20 questions was sent to 60 patients of a single neurosurgical center who had undergone surgery for an intramedullary spinal cord tumor. A retrospective study was carried out based on this data. Histological findings ranged from ependymoma (52.9%), hemangioblastoma (17.6%), cavernoma (14.7%), astrocytoma (8.8%), oligodendroglioma (2.9%) to ganglioglioma (2.9%). Non-scientific therapy was defined as a method not used in routine clinical practice for the treatment of symptoms induced by intramedullary spinal cord tumors. RESULTS: A total of 38 questionnaires was returned. About 55.3% of the patients claimed to use non-scientific therapies. No significant difference between histological types and the percentage of alternative therapy use was detected. There was a gender difference. One third (non user group) to one fourth (user group) did not feel adequately informed about their disease. The monthly costs for non-scientific therapies ranged from 50 to 500 Euros. The main motive for the use of non-scientific therapies was the wish to try everything possible. CONCLUSION: In the daily clinical routine, patients' use of non-scientific therapies may be largely overlooked and underestimated. Neurooncologists should be aware of this phenomenon and encourage an open but critical dialogue with their patients.

Training microneurosurgery - four years experiences with an in vivo model.

OBJECTIVE: Enquiries among surgical trainees revealed an increasing discontent regarding their quality of training. 40 % of young surgical trainees judge their training as inadequate and 70% are offered no structured training programme. Working time restrictions and economic pressure may be strong factors hindering residents from becoming skillful surgeons. Therefore, additional forms of training seem to be needed. METHOD: An in vivo swine model was evaluated for its practical use in training neurosurgical residents. Surgical procedures included craniotomy, dural opening, brain surgery and excision of an artificial tumour created by injection of coloured fibrin glue. Microscopy and bleeding management with bipolar cautery and haemostyptics were an integrated part of training. Supervision by experienced neurosurgeons with up to 3 trainees in a 2-day course was warranted. Standardised questionnairies before and after training were used to assess the quality and utility of the programme. RESULTS: 24 residents have participated in the course (1 (st)-5 (th) year of training). Minor experience with less than 100 conducting surgeries was seen in 59% of trainees. 14 residents had participated in more than 100 surgeries as first assistant. Spinal surgery was the predominant common experience. All participants judged their surgical training as insufficient. 77% had no microsurgical lab at their clinics. Expectations for the course were met for all trainees and the tutorials judged as excellent (65%) or good (35%). Positive evaluations of the in vivo model (97%), a realistic laboratory setup (94%), the working environment (94%) and close supervision (94%) showed that these were the most favourable aspects of the course. CONCLUSION: Educational training in surgical specialities is becoming a major problem in our daily practice and requires additional training facilities. In this context, in vivo models are an ideal opportunity for young neurosurgeons to train bleeding management and surgical complications in particular. This educational form is thought to be a unique training model which is now added by spinal and neurovascular courses.

Complementary therapy use in patients with glioma: an observational study.

OBJECTIVE: Despite novel multimodal therapeutic approaches, the vast majority of glial tumors are not curable. Patients may search for complementary therapies in order to contribute to the fight against their disease or to relieve symptoms induced by their brain tumor. The extent of the use of complementary or alternative therapies, the patients' rationale behind it, and the cost of complementary therapy for gliomas are not known. We used a questionnaire and the database of the German Glioma Network to evaluate these questions. METHODS: A total of 621 questionnaires were available for evaluation from patients with glial tumors of WHO grades II to grade IV. The patients were recruited from 6 neuro-oncologic centers in Germany. Complementary therapy was defined as methods or compounds not used in routine clinical practice and not scientifically evaluated. RESULTS: Forty percent of the responding patients reported the use of complementary therapies. Significant differences between the group of complementary therapy users and nonusers were seen with respect to age (younger > older), gender (female > male), and education (high education level > low education level). The motivation for complementary therapy use was not driven by unsatisfactory clinical care by the neuro-oncologists, but by the wish to add something beneficial to the standard of care. CONCLUSIONS: In clinical practice, patients' use of complementary therapies may be largely overseen and underestimated. The major motivation is not distrust in conventional therapies. Neuro-oncologists should be aware of this phenomenon and encourage an open but critical dialogue with their patients.

Intracranial aneurysms in childhood: report of 8 cases and review of the literature.

OBJECTIVE: Subarachnoid haemorrhage (SAH) due to ruptured cerebral aneurysms is a rare diagnosis in childhood and is believed to differ from that in adults with regard to sex, aneurysm size and location, clinical appearance as well as outcome, suggesting that aneurysms in childhood are a distinct entity. PATIENTS AND METHODS: Seven children and young adults with a mean age of 13.6 years suffered from aneurysmal subarachnoid haemorrhage. On admission two patients were Hunt & Hess (HH) grade I, four were grade III and one was grade V. In addition, there was one 14-year-old boy who presented with seizures and was found to have a non-ruptured aneurysm of the basilar tip. Overall, there were two female and six male patients. Anterior circulation aneurysms were diagnosed in five patients whereas three patients suffered from posterior circulation aneurysm. Five large (10-25 mm) aneurysms and three smaller than 10 mm were found. Surgical clipping of the aneurysms was performed in all patients. RESULTS: Six patients showed no significant disability or no symptoms at all when evaluated with the modified Rankin Scale (mRS 0 or 1). One patient suffered from persisting nerve palsies (mRS 2) and one 6-year-old boy, admitted with HH grade V, died due to the initial poor clinical condition followed by severe brain oedema and secondary complications. CONCLUSIONS: Reviewing the literature, a male predominance, low rate of SAH, high percentage of complex aneurysms and of aneurysms located in the posterior circulation are characteristic features of aneurysms in childhood. Once a cerebral aneurysm is diagnosed in the younger age group, definite therapy should be performed soon as the outcome is expected to be more favourable than in adulthood.

[Prognosis after surgical treatment of spinal metastases due to lung cancer]. / Prognose nach operativem Wirbelsäuleneingriff bei Bronchialkarzinommetastasen.

BACKGROUND: Patients with spine metastases due to lung cancer suffer from a reduced lifespan. For a more precise prognosis, it is important to define parameters which influence the individual survival time. This study reviewed the mean survival time of patients who had undergone surgery because of spine metastases due to lung cancer. It should be evaluated whether the postsurgical survival time is dependent on the length of time between diagnosis and surgery and from the histological type of the tumor. MATERIAL AND METHODS: Between January 1999 and December 2003, 68 patients had undergone spine surgery because of spine metastases due to lung cancer at the department of traumatology of the St. Georg General Hospital in Hamburg, Germany. Retrospective data were collected from the hospital documentary system regarding the period between diagnosis of lung cancer and date of surgical treatment, and regarding the histological type of the tumor. The postsurgical survival times were evaluated using data from the Hamburger Cancer Index and from general practitioners. These times were analysed afterward according to the defined parameters. RESULTS: The average age was 62.6 years, and 24 female and 44 male patients were included. It was possible to evaluate the survival time of 65 patients. The average survival time of those with preoperative unknown primary manifestation (20 patients) was 88 days, in the group of patients with lung cancer histories of less than 12 months (35 patients) 141 days, and with patients with lung cancer histories of lung cancer longer than 12 months (13 patients) it was 171 days. The mean survival times after surgical treatment were 122 days for patients suffering from non-small-cell lung cancer (45 patients), 128 days for those with small-cell lung cancer (20 patients), and 247 days for patients with other histological types (three patients). DISCUSSION: The prognosis of patients after spine surgery for lung cancer metastases is poor. The indication for surgical treatment of spinal metastases due to lung cancer should be critically discussed. Especially patients with unknown primary manifestation might benefit from a conservative approach. With respect to the patients' reduced lifespan, they, their relatives, and the nursing staff should be carefully informed.

Disarrangement of fiber tracts and decline of neuronal density correlate in glioma patients--a combined diffusion tensor imaging and 1H-MR spectroscopy study.

BACKGROUND AND PURPOSE: Diffusion tensor imaging (DTI) and MR spectroscopy are noninvasive, quantitative tools for the preoperative assessment of gliomas with which the quantitative parameter fractional anisotropy (FA) and the concentration of neurometabolites N-acetylaspartate (NAA), choline (Cho), creatine (Cr) of the brain can be determined. Measurements of FA and NAA reflect the integrity of fiber tracts and the presence of neurons, respectively. This investigation examines changes of FA and NAA and compares these different aspects in architecture of gliomas after spatial coregistration. METHODS: DTI and chemical shift (1)H-MR spectroscopy was performed in 34 healthy volunteers and 69 patients with histologically confirmed (n = 48) or morphologically suspected (n = 21) non-necrotic brain glioma. Volumes of interest (VOIs) were placed in the tumor center (TC), the tumor border (TB), the normal-appearing white matter adjacent to the tumors (TNWM), and in the white matter of the contralateral hemisphere (NWMC). Median FA values and NAA/Cr and NAA/Cho ratios were calculated in the patients' VOIs and the gray and white matter of the volunteers. Correlations of FA values and NAA ratios were calculated. RESULTS: Continuous changes of FA and NAA from the tumor center to the periphery (the adjacent white matter and the contra-lateral hemisphere, respectively) were observed, where median values were: TC: 0.73 +/- 0.45, 0.47 +/- 0.58, 0.17 +/- 0.15 (NAA/Cr, NAA/Cho, FA); TB: 1.06 +/- 0.53, 1.00 +/- 0.15, 0.23 +/- 0.08; TNWM: 1.42 +/- 2.48, 1.21 +/- 0.95, 0.34 +/- 0.09; and NWMC: 1.63 +/- 0.72, 1.56 +/- 1.34, 0.38 +/- 0.08. Correlation of median FA values and NAA ratios in the cumulative group of patients was high (r = 0.99 [NAA/Cr], 0.95 [NAA/ Cho] at P < .01). Correlation between the individual NAA ratios and the FA values was moderate (r = 0.53 [NAA/Cr], 0.51 [NAA/Cho] at P < .01). CONCLUSION: In gliomas, the degree of tissue organization decreases continuously from the surrounding tissue toward the center of the tumor accompanied by a concordant decrease of NAA. This uniform behavior of FA and NAA reflects a decreasing integrity of both neuronal structures and fibers.

Hollow mandrin facilitates external ventricular drainage placement.

BACKGROUND: Placement of ventricular catheters is a routine procedure in neurosurgery. Ventricle puncture is done using a flexible ventricular catheter stabilised by a solid steel mandrin in order to improve stability during brain penetration. A correct catheter placement is confirmed after removing the solid steel mandrin by observation of cerebrospinal fluid (CSF) flow out of the flexible catheter. Incorrect placement makes further punctures necessary. The newly developed device allows CSF flow observation during the puncture procedure and in addition precise intracranial pressure (ICP) measurement. METHOD: The developed mandrin is hollow with a blunt tip. On one side 4-5 small holes with a diameter of 0.8 mm are drilled corresponding exactly with the holes in the ventricular catheter, allowing CSF to pass into the hollow mandrin as soon as the ventricle is reached. By connecting a small translucent tube at the distal portion of the hollow mandrin ICP can be measured without loss of CSF. The system has been used in 15 patients with subarachnoid haemorrhage (SAH) or intraventricular haemeorrhage (IVH) and subsequent hydrocephalus. FINDINGS: The new system improved the external ventricular drainage implantation procedure. In all 15 patients catheter placement was correct. ICP measurement was easy to perform immediately at ventricle puncture. In 4 patients at puncture no spontaneous CSF flow was observed, therefore by connecting a syringe and gentle aspiration of CSF correct placement was confirmed in this unexpected low pressure hydrocephalus. Otherwise by using the conventional technique further punctures would have been necessary. CONCLUSIONS: Advantages of the new technique are less puncture procedures with a lower risk of damage to neural structures and reduced risk of intracranial haemorrhages. Implantation of the ventricular catheter to far into the brain can be monitored and this complication can be overcome. Using the connected pressure monitoring tube an exact measurement of the opening intracranial pressure can be obtained performed without losing CSF.

Hydrocephalus therapy: reduction of shunt occlusions using a peel-away sheath.

In order to avoid shunt occlusions through particles of brain parenchyma a new procedure was used. Conventionally during ventricle puncture brain tissue may intrude into perforating holes of the ventricular catheter and subsequently shunt dysfunction may occur. By using a peel-away sheath the ventricular catheter can be protected during puncture. The conventional technique (n=90) was compared with the peel-away sheath technique (n=20) in a retrospective analyses in regard to shunt revisions during a 1-year period. Shunt revision was done in 18% (16/90) within 1 year using the conventional technique, respectively, 5% (1/20) using the peel-away sheath technique. Although criteria for statistical significance were not reached these data are worth to be mentioned. A randomised prospective study is initiated and the key features are presented.

Diffuse arachnoidal enhancement of a well differentiated choroid plexus papilloma.

INTRODUCTION: The case of a 38-year old man with a histologically benign choroid plexus papilloma arising within the fourth ventricle with en plaque growth around the brain stem and medulla is described in detail. Up to this point this particular growth pattern has not been published and is a rare presentation for this tumour. CLINICAL PRESENTATION: The patient presented with a 1.5 year history of headache, nausea, and vomiting in the morning. Additional symptoms like blurred vision and gait ataxia lead to hospital admission. MRI demonstrated a homogeneously contrast-enhancing tumour completely filling the fourth ventricle and subsequent obstructive hydrocephalus. In addition Gd enhancement encasing the brain stem, the lower aspect of the medulla and the conus medullaris was seen suggesting a disseminated ependymoma or medulloblastoma. INTERVENTION: An extensive resection of the tumour in the fourth ventricle and CP angle was performed. Infiltrative growth into the structures of the left CP angle and into the rhomboid fossa hampered complete removal. Surprisingly histological examination revealed a well-differentiated papillary choroid plexus papilloma without signs of anaplasia. On follow up imaging the Gd enhancement encasing the pons vanished completely. A growing cyst adjacent to a small tumour residuum left behind on the floor of the fourth ventricle led to re-operation after 8 months with complete removal. DISCUSSION: This case presents several biological, neuroradiological and surgical aspects which make it noteworthy and we hope that the informations provided add to the understanding of these tumours, expand the differential diagnostic thinking of lesions which present with diffuse arachnoid Gd enhancement upon first presentation.

Expression of cyclooxygenase 2 (COX-2) in human glioma and in vitro inhibition by a specific COX-2 inhibitor, NS-398.

The up-regulation of cyclooxygenase 2 (COX-2) expression is a frequent occurrence in a variety of different tumors. In this study, COX-2 protein expression was investigated in 50 glioma and 3 normal brain specimens by immunohistochemistry. Expression of COX-2 protein was observed in all normal brain and glioma specimens by immunohistochemistry, regardless of histological grade. The immunoreactive score was significantly higher in high-grade glioma than low-grade glioma and normal brain specimens. For a subset of these tumors (nine gliomas and three normal brain), Western blot analysis was also performed. COX-2 protein was detected in all specimens by Western blot analysis. The effect of the specific COX-2 inhibitor NS-398 on monolayer cell cultures and three-dimensional glioma spheroids was investigated using U-87MG and U-251MG human glioblastoma cell lines. The proliferation rate was assessed in monolayer cultures. In addition, a growth assay, a migration assay, an apoptosis assay, and a tumor invasion assay were performed in a three-dimensional spheroid culture system. NS-398 was able to reduce the proliferation of monolayer cell cultures, as well as the growth of spheroids and tumor cell migration, in a dose-dependent manner. There was also a moderate increase in the number of apoptotic cells in the treated spheroids. NS-398 did not have an inhibitory effect on tumor invasion in the coculture spheroid system. Our study provides evidence that COX-2 is up-regulated in the majority of high-grade gliomas and that a potential role of COX-2 inhibitors as an adjuvant therapy for brain tumors may exist.

Regulation of secretion of PTHrP by Ca(2+)-sensing receptor in human astrocytes, astrocytomas, and meningiomas.

Parathyroid hormone-related protein (PTHrP) is the major mediator of the humoral hypercalcemia of malignancy and of malignant osteolysis associated with skeletal metastases of common epithelial cancers. PTHrP secretion is regulated by the extracellular calcium concentration ([Ca(2+)](o)) in several types of normal and malignant cells. Because the [Ca(2+)](o)-sensing receptor (CaR) is a key mediator of [Ca(2+)](o)-regulated hormone secretion [e.g., of parathyroid hormone (PTH) by parathyroid chief cells], we investigated the expression of the CaR and PTHrP in normal and neoplastic glial cells and studied the effects of [Ca(2+)](o) on PTHrP secretion. Our results show that primary embryonic human astrocytes (HPA) express CaR mRNA and protein as detected by RT-PCR and Western analysis, respectively. Furthermore, astrocytomas and meningiomas also express the CaR at similar levels as assessed by RT-PCR and Northern and Western blot analyses. HPA and astrocytomas express transcripts encoding all three known isoforms of PTHrP [PTHrP(139), PTHrP(141), and PTHrP(173), comprising 139, 141, and 173 predicted amino acid residues, respectively] as assessed by RT-PCR, whereas meningiomas express only the first two of these. Finally, elevated levels of [Ca(2+)](o) and other polycationic CaR agonists dose dependently stimulate PTHrP secretion from HPA, astrocytomas, and meningiomas, although both basal and high [Ca(2+)](o)-stimulated rates of PTHrP secretion are approximately 2. 5-fold higher in HPA than in the glial tumors studied here. Therefore, our results show that HPA, astrocytomas, and meningiomas express both the CaR and PTHrP and that CaR agonists stimulate PTHrP secretion.