Differential screening-selected gene aberrative in neuroblastoma (DAN) is increased in the CSF of patients with MS and may be induced by therapy with interferon-ß.

Bone morphogenic proteins (BMPs) signaling blockade induce neurogenesis and oligodendrogenesis. Differential screening-selected gene aberrative in neuroblastoma (DAN) is a glycoprotein that antagonizes BMPs. We found that DAN levels were higher in CSF compared to serum in all participants. CSF-DAN levels were elevated in RR-and progresssive MS patients compared to controls. Moreover, serum-DAN levels were reduced in those patients, but elevated in IFN-ß1a treated patients. The main source of DAN is apparently CNS- resident cells. The enhanced levels of CSF-DAN in MS patients suggest a tendency to induce neurogenesis/oligodendrogenesis in the patients CNS. Our results suggest an unreported mode of action of IFN-ß1a.

Chronic exposure to cerebrospinal fluid of multiple system atrophy in neuroblastoma and glioblastoma cells induces cytotoxicity via ER stress and autophagy activation.

Oncogenesis and neurodegeneration share many common pathogenic pathways, involved in endoplastic reticulum (ER) stress, autophagy, DNA repair, and oxidative stress. However, mechanisms of cross-talking between oncogenesis and neurodegeneration are still unknown. Characterized by abnormal accumulation of α-synuclein (α-syn) aggregates in central nervous system (CNS), multiple system atrophy (MSA) is classified as α-synucleinopathy. Rapidly emerging evidence suggests that 'prion-like propagation' of α-syn aggregates in the regional spread of CNS leads to the progression of α-synucleinopathy. Whether cerebrospinal fluid (CSF) has deteriorating effects on neurogenic tumor cells and is involved in progression of α-synucleinopathy has not been explored. Here, we first show the cytotoxic effects of MSA-CSF on the neuroblastoma and glioblastoma cells and its underlying mechanism in vitro. Remarkably, MSA-CSF induced cytotoxicity via activating ER stress-associated apoptosis and autophagy in both SH-SY5Y and U251 cells. The result from in vivo systematic neuropathological analysis reveals that abnormally activated ER stress and autophagy were confined to substantia nigra and cerebellum in mouse CNS following MSA-CSF treatment. Specifically, dopamine neurons in substantia nigra and Purkinje cells in cerebellum cortex were degenerated in MSA-CSF-injected mice. Altogether, these findings demonstrate that MSA-CSF exerts cytotoxicities on nervous system neoplasms and accelerates the progression of synucleinopathies.

Diagnostic identification of malignant cells in the cerebrospinal fluid by tumor-specific qRT-PCR.

Tumor specific quantitative RT-PCRs for two neuroblastoma specific molecular markers, tyrosine hydroxylase (TH) and GD2 synthase, were used to unequivocally demonstrate the neoplastic nature of the cells present in the cerebrospinal fluid of a neuroblastoma patient. After radical surgery of two separate tumoral lesions, localized in the extradural area, the patient presented with meningitis. Common sites of neuroblastoma metastatization, e.g. bone and bone marrow, were not infiltrated by tumor cells, as assessed by standard scintigraphy, morphological investigation and by sensitive and specific immunocytochemical and molecular assays. The results presented here demonstrate the successful use of tumor-specific qRT-PCRs in cerebrospinal fluid to investigate questionable clinical cases. The technique, which compared to other detection methods (e.g., immunocytochemistry) requires very few cells, yields unambiguous information once a suspected diagnosis has been formulated and a tumor-specific molecular marker is available.

Evidence of cellular immune activation in children with opsoclonus-myoclonus: cerebrospinal fluid neopterin.

To evaluate cellular immune activation in opsoclonus-myoclonus syndrome, we measured the inflammatory marker neopterin in the cerebrospinal fluid of 16 children with opsoclonus-myoclonus and neuroblastoma, 24 children with opsoclonus-myoclonus but no tumor, and 19 age-matched controls. The mean concentration in opsoclonus-myoclonus was 2.3-fold higher than in controls (P = .008). Neopterin was greatly elevated in four of the most neurologically severe cases, up to 8.3-fold above the highest control level. Thirteen of the 40 children with opsoclonus-myoclonus but no controls had a neopterin concentration >2 SD above the control mean (P = .005). In this high neopterin subgroup, neurologic severity was significantly greater and the duration of neurologic symptoms was less. In 16 children re-examined on immunotherapy, including adrenocorticotropic hormone (ACTH) combination therapy, treatment was associated with a significant reduction in both neopterin and neurologic severity. Neopterin did not differ significantly between the tumor and non-tumor opsoclonus-myoclonus etiologies. No abnormalities of tetrahydrobiopterin were found. Although cerebrospinal fluid neopterin lacked the sensitivity to be a biomarker of disease activity in opsoclonus-myoclonus, elevated concentrations do support a role for T-cell activation and cell-mediated immunity in its pathophysiology.

Treatment of neoplastic meningeal xenografts by intraventricular administration of an antiganglioside monoclonal antibody, 3F8.

Leptomeningeal (LM) neoplastic metastases are painful, debilitating and inevitably lethal. Intrathecal (IT) anti-tumor antibodies may have therapeutic potential. We evaluated 3F8, an anti-G(D2) murine IgG(3) monoclonal antibody (MAb) in the treatment of human melanoma (SKMEL-1) and neuroblastoma (NMB7) xenografts in athymic rats. Both tumors were lysed efficiently in vitro by 3F8 in the presence of rat neutrophils or rat complement. Antibody-dependent cellular cytotoxicity (ADCC) was not augmented by recombinant human GM-CSF (rhGM-CSF), rhG-CSF, recombinant rat MIP-2 (rrMIP-2) or lipopolysaccharide (LPS). In vivo, continuous intraventricular administration of 3F8 and LPS prevented tumor engraftment, retarded tumor growth and eradicated 3-day-old established xenografts whereas 3F8 alone, LPS alone or F(ab)'(2) plus LPS had no or only marginal effects. Tumor establishment in brain was completely prevented in 36% of animals implanted with SKMEL-1 and 65% of animals implanted with NMB7. Twenty percent of established xenografts around the brain were eradicated but all animals had persistent tumor in the lumbosacral meninges despite treatment. Continuous intraventricular infusion of LPS produced a variable polymorphonuclear (PMN) pleocytosis that was dose-dependent. Continuous intraventricular infusion of 3F8 produced immunohistochemically detectable attachment to 86% of persistent brain deposits of tumor but <1% of spinal lumbosacral deposits. We conclude that regional therapy with anti-G(D2) MAb could target neutrophils to inhibit LM tumor growth. However, optimal activation and mobilization of neutrophils into the cerebrospinal fluid (CSF) and improved penetration of MAb to tumor sites remain critical variables.

Antineuronal antibody in a patient with neuroblastoma and opsoclonus-myoclonus-ataxia: a case report.

We describe a case of a patient with neuroblastoma and opsoclonus- myoclonus ataxia displaying serum and CSF anti-Hu antibodies that were able to recognize antigens of the patient's own tumor.

Differential spectrum of expression of neural cell adhesion molecule isoforms and L1 adhesion molecules on human neuroectodermal tumors.

A series of four medulloblastomas, seven neuroblastomas (Nb), two ependymomas, and three gliomas, human neuroectodermal tumors, were screened for their expression of adhesion molecules L1, carcinoembryonic antigen, neural cell adhesion molecule isoforms (N-CAM) and HNK1 epitope by Western blotting and double immunofluorescence labeling. All seven neuroblastomas, whatever their differentiated state, expressed L1, a neural cell surface developmental antigen, whereas all other tumors tested were negative. All tumors expressed N-CAM; however, a large diversity was observed among the isoforms. Low sialylated N-CAM 140 was present, with different intensity, in ependymomas and astrocytomas. High sialylated isoforms were detected by a monoclonal antibody (anti-MenB) specifically recognizing high polymers of alpha 2-8 linked neuraminic acid. They were expressed in all medulloblastomas studied (4 of 4), and in 4 of 7 Nbs examined. Negative cases corresponded to tumors having undergone chemotherapeutic treatment or to ganglioneuroma. The interconversion from high to low sialylated forms might reflect changes which are critical for the conversion of Nbs into benign ganglioneuromas. HNK1 epitope was expressed on a large diversity of molecules by nearly all tumors except two Nbs which were also negative with anti-MenB antibody. This simultaneous loss of carbohydrate epitopes could correlate with higher maturation states of the tumors. None of the tumors expressed carcinoembryonic antigen. Therefore, anti-L1 and anti-MenB antibodies define differentiation-related antigens that could differentiate between Nbs and other tumors and may prove helpful in diagnosis and understanding of the biological nature of neuroectodermal tumors. An immunodot assay was set up and allowed to titrate the presence of polysialic acid units in cerebrospinal fluid from patients presenting meningeal spread of medulloblastomas. It could help to assess metastasis and to monitor the effects of chemotherapeutic treatment on polysialylated N-CAM positive tumors.

Elevation of cerebrospinal fluid catecholamine metabolites in patients with intracranial tumors of neuroectodermal origin.

The concentrations of homovanillic acid (HVA), hydroxymethoxyphenylethyleneglycol (HMPG), and vanillylmandelic acid (VMA) were determined in lumbar cerebrospinal fluid (CSF) specimens. The study population consisted of the following groups: control subjects with malignancies of nonneuroectodermal origin (mostly leukemia in remission), neuroblastoma (extracranial and intracranial or cranial metastases), brain tumors (neuroectodermal and glial), and retinoblastoma. A significant increase in the CSF concentration of HVA was observed in patients with brain tumors (neuroectodermal), neuroblastoma (intracranial or cranial metastases), and retinoblastoma when compared with age-matched controls. In contrast, HMPG and VMA concentrations did not differ from controls except in patients with neuroblastoma (intracranial or cranial metastases) and brain tumors (neuroectodermal) who had significant elevations in HMPG and VMA, respectively. An inverse correlation was noted between the CSF concentration of HVA and clinical response to therapy. Nonresponding patients exhibited increases in HVA when compared with pretreatment values. These data suggest that the quantitative determination of catecholamine metabolites in lumbar CSF is an effective method for diagnosing intracranial tumors of neuroectodermal origin and assessing their response to therapy.

[Cytologic examination of the cerebrospinal fluid in the diagnosis of primary or metastatic neoplasms of the nervous system]. / Exame citológico do líquido céfalo-raquidiano no diagnóstico de neoplasias primitivas ou metastáticas do sistema nervoso: a propósito de seis casos.

The authors report 6 cases of cerebrospinal fluid (CSF) examinations in which malignant cells were found. In 5 cases the finding was incidental: medulloblastoma (case 1); malignant melanoma (case 2); adenocarcinoma (case 3); meningitis carcinomatosa (case 4) and neuroleukaemia (case 5). In only one case neuroleukaemia was suspected before the study of the CSF (case 6). The unexpected occurrence of malignant cells in the CSF demands the pathologist to be well acquainted with tumor cell cytology, in order to identify them providing so useful information that can decidedly influence subsequent diagnostic and therapeutic procedures. The cell collection technic developed in the authors' laboratory was considered adequate, because both inflammatory and malignant cells were securely identified. Its principle is to obtain thin air-dried smears that dry almost instantaneously on slides, in order the cellular structure be preserved. After centrifuging about 5-8 ml of CSF the tube is inverted so as to pour off the whole of the supernatant fluid. It is kept inverted with the operator's left hand, so that no drop of fluid can run back on to the cells. With his right hand the operator touches the bottom of the inverted tube with a Pasteur micropipette preciously made by distending a microhematocrit tube under the flame of a Bunsen burner and attached with adhesive tape to the handle of a wire loop 2 mm in diameter.(ABSTRACT TRUNCATED AT 250 WORDS)

Exame citológico do líquido céfalo-raquidiano no diagnóstico de neoplasias primitivas ou metastáticas do sistema nervoso / Cytological examination of cerebrospinal fluid in the diagnosis of primary or metastatic neoplasms of the nervous system

Relata-se o encontro de células neoplásicas em 6 casos de exames de LCR realizados na rotina do laboratório. Em 5 casos o encontro dessas células foi acidental: meduloblastoma (caso 1); melanoma (caso 2); adenocarcinoma (caso 3); meningite carcinomatosa (caso 4) e neuroleucemia (caso 5). Em um caso havia suspeita clínica de neuroleucemia (caso 6). A ocorrência imprevisível de células neoplásicas no LCR exige que o liquorologista domine a citologia oncótica, para que näo sejam perdidas informaçöes essenciais para o diagnóstico, prognóstico e terapêutica do caso em estudo. A técnica para a obtençäo de preparados destinados ao estudo da citologia do LCR foi desenvolvida no laboratório dos autores e mostrou-se adequada, por permitir o reconhecimento de células inflamatórias e de células neoplásicas com igual segurança. O fundamento da técnica descrita consiste na preparaçäo de esfregaços de pequena extensäo, muito finos que secam quase instantaneamente ao ar, permitindo assim que a estrutura das células seja conservada. Para tanto o sedimento resultante da centrifugaçäo de 5-8ml de LCR é suspenso no pouco de LCR que sobra no tubo após o sobrenadante ter sido transferido para o outro tubo. O tubo que contém o sedimento é mantido invertido com a mäo esquerda do operador, que, com a direita, toca o fundo do tubo com micropipeta de Pasteur, feita pela distençäo de um tubo de microhematócrito, previamente colado ao cabo de uma alça de platina com 2mm de diâmetro. Uma gota da suspensäo de células é soprada com pipetador hematológico sobre uma lâmina e distendida com alça, à moda de "pente" numa área máxima de 1cm2. Após secagem por agitaçäo ao ar, o preparado é corado pelo Giemsa ou Leishman

Tumor markers and cytologic features of cerebrospinal fluid.

Tumor markers are useful in establishing the diagnosis of certain central nervous system tumors, especially germinal tumors of the pineal region. They are not sufficiently specific to be able to replace biopsy for exact diagnosis. They may also be useful for monitoring of therapy, as an indicator of recurrence of the tumor. Cerebrospinal fluid cytology is not generally useful in establishing a specific histologic diagnosis, especially in children, but can help to monitor therapy and predict tumor recurrence. More extensive studies are needed in both areas to define more precisely the role of markers and cytologic studies.

Cytology of ganglioglioma of the brain: a morphometric study.

This study describes and illustrates the application of the morphometric method in the classification of different types of cells in mixed populations. The nuclei of neoplastic cell and astrocytes, nonneoplastic monocytoid cells and lymphocytes in touch preparations of two gangliogliomas were measured. Two preoperative cerebrospinal fluid specimens from one of these cases were similarly evaluated. The mean nuclear area of some cell populations was statistically distinct and consistent in each preparation. Other cell populations showed considerable overlap in nuclear size and could not be separated on the basis of this parameter alone.

DOPA metabolism in neuroblastoma.

Blood plasma samples from 60 neuroblastoma patients prior to, during, and following treatment were studied for their content of circulating DOPA using a radioenzymatic assay. Normal values were established from children who were tumor-free or had other nonneurogenic tumors. The highest plasma DOPA concentration in tumor-free or nonneuroblastoma controls was 5.3 ng/ml with a mean of 2.15 ng/ml. Most neuroblastoma patients (28/31) with active disease had DOPA values above this level. Only one out of 30 "successfully" treated patients without evidence of disease was encountered with an abnormally high level. In treated patients, elevated values forewarned of impending clinical recurrence or persistent tumor. Cerebrospinal fluid DOPA levels in one patient with cerebral neuroblastoma were extraordinarily high and suggests that this assay may prove useful to distinguish neuroblastoma from other central neuroectodermal or metastatic tumors. Plasma DOPA appears to be a reliable predictive and diagnostic test for neuroblastoma.

[Cytological changes in the csf caused by inthathecal administration of methotrexate. II. Studies on a case of neuroblastoma with cerebellar symptoms in a child]. / Liquorcytologiai elváltozások intrathecalis methotrexat kezelés hatására. II. Vizsgálatok kisagyi tünetekkel kísért gyermekkori neuroblastoma esetében

A case of an olfactory neuroblastoma of a 5 years old child is reported. The tumour propagated into the intracranial space and produced cerebellar symptoms. Features of the neuroblastic cells, found in the cerebrospinal fluid and their alterations as an effect of the intrathecal therapy with Mathotrexat are analysed. Authors emphasize the differences of alterations of leukaemic and neuroblastic cells called into being by the Methotrexat therapy.