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Background/aim: Brain metastasis, a leading cause of cancer death, is a clinical challenge. Recently, genetic characterization of brain metastatic lesions based on next generation sequencing-based advanced technologies, such as single-cell RNA sequencing, has been performed to develop novel efficient therapies. The present study aimed to investigate brain-metastasis-specific biomarkers as well as relevant prognostic factors., Patients and Methods: The genetic profiles and expression levels of immune response-associated genes and 820 cancer-associated genes were compared between primary cancer lesions and metastatic cancer lesions obtained from nine cancer patients at the Shizuoka Cancer Center. Cytokine and chemokine marker genes were analyzed via quantitative PCR. T-cell receptor (TCR) repertoire profiling was performed for the same patients. For survival analysis, survival data of 52 cancer patients with brain metastases were utilized., Results: Comparison of driver mutation profiling between primary and metastatic lesions revealed shared core mutations in both lesions and a few new mutations in metastatic lesions. A high tumor mutation burden (TMB) was detected in metastatic lesions. Volcano plot analysis revealed specific features of the metastatic tumor microenvironment, such as cancer signaling promotion and immune suppression due to decreased immune cell infiltration. Survival analysis revealed that three genes, the TREML2 gene, the BTLA gene on activated microglia and the CERS2 gene on metastatic tumor, were potent prognostic factors., Conclusion: High TMB in metastatic lesions indicates potential benefit from immune checkpoint inhibitor usage for brain metastasis and TREML2 and BTLA are factors associated with poor prognosis. Activated microglia may be novel targets for the treatment of brain metastasis., (Copyright © 2024 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Many attempts to reduce radiation fields for intracranial germ cell tumors (iGCTs) remain unsuccessful. To assess the possibility of reduction, we analyzed registry data of 57 patients who mostly underwent local irradiation for iGCTs between 1997 and 2006. The recommended treatment for pure germinomas (PGNs) included 3 courses of cisplatin and etoposide followed by 24 Gy local irradiation. Intensified chemotherapy using a combination of cyclophosphamide and intrathecal methotrexate was recommended for human chorionic gonadotropin-producing germinomas (hCG-GNs) and nongerminomatous germ cell tumors (NGGCTs); both received 50.4 Gy local irradiation. High-dose chemotherapy was only administered for residual NGGCTs after chemoradiotherapy. Craniospinal irradiation was recommended only in metastatic cases. During the median follow-up of 114.8 months, 8 of 9 relapses from 24 PGNs occurred outside irradiation fields, with a 5-year progression-free survival (5-year PFS) of 75%±8.8%. Conversely, no recurrences occurred from 11 hCG-GNs, with a 5-year PFS of 100%. Eleven of 22 patients with NGGCTs received high-dose chemotherapy; the 5-year PFS was 81.3%±8.4%; 2 of 3 relapses occurred in the spinal cord. Thus, local irradiation for PGNs was insufficient without treatment intensification. The introduction of intensified chemotherapy improved outcomes of both patients with hCG-GNs and NGGCTs. However, the contributions of either modality remained unclear.

Langerhans cell histiocytosis (LCH) is sometimes resistant to conventional chemotherapies, and treatment with 2-chlorodeoxyadenosine (2-CdA) is gaining importance as a salvage treatment for refractory or recurrent LCH. Secondary malignancies such as acute myelogenous leukemia and myelodysplastic syndrome (MDS) due to 2-CdA have recently been reported. However, there have been no reports to date of cases of 2-CdA-related secondary MDS in which spontaneous remission was achieved. Here, we report the case of a 1-year-old boy with an occipital tumor who was diagnosed with LCH by biopsy and underwent chemotherapy. However, the disease relapsed and became refractory to chemotherapy. He received 2-CdA treatment, which was effective. However 6 months after the start of treatment, he developed MDS with chromosomal abnormality of 7q-. After 1-year observation without any intervention, his hematological findings had returned to normal, and the chromosomal abnormality had also disappeared. To our knowledge, this is the first report of 2-CdA-related MDS with spontaneous hematological remission.

We report two cases of ependymoma which showed prominent "granular cell" changes of the cytoplasm. The patients were a 7-year-old boy with a tumor in the cerebellum (case 1) and a 70-year-old man with a tumor in the frontal lobe (case 2). The tumor of case 1 showed a histopathological appearance of ependymoma containing many focal aggregates of large polygonal cells in which the cytoplasm was stuffed with numerous eosinophilic granules. The tumor of case 2 predominantly showed the features of papillary ependymoma, and some tumor cells were swollen and contained similar eosinophilic granules. Intracytoplasmic granules in both tumors were immunoreactive for GFAP and ubiquitin, but not for epithelial membrane antigen, CD68 or mitochondria. Ultrastructurally, they were found as aggregates of membrane-bound, electron-dense, globular structures. Karyotypic analysis of the tumor in case 1 demonstrated 2, 11 and 12 trisomies. Intracytoplasmic eosinophilic granules occasionally occur in astrocytic and oligodendroglial neoplasms, but an appearance of similar granules is very rare in ependymoma. The two cases presented here may represent a new histopathological variant of ependymoma, and the term "granular cell ependymoma" is appropriate for them., (© 2012 Japanese Society of Neuropathology.)

The authors present a pediatric case of ganglioglioma occurring in the medulla oblongata. A 7-year-old boy was referred to our hospital with complaints of ataxia, seizure and sleep apnea. MRI of the brain disclosed a large tumor occupying the medulla oblongata, and the upper portion of the cervical spinal cord was also involved. The patient underwent midline suboccipital craniotomy and laminectomy of C1 to attempt radical resection of the tumor, which resulted only in partial removal of the tumor due to severe bradycardia during the operation. The histological diagnosis was ganglioglioma, WHO grade 2. Although both radiotherapy and chemotherapy were performed following the operation, the tumor remained unchanged. The patient died of respiratory arrest five months after the operation. Gangliogliomas usually occur in the supratentorial region, which permits easy surgical access and good prognosis. Only 3% of gangliogliomas occur in the brain stem, and its management can be challenging because of the difficulty of radical resection and poor response to both radiotherapy and chemotherapy.

A tectal glioma presenting with late-onset aqueduct stenosis and obstructive hydrocephalus is usually categorized as a benign glioma. Apparent clinical or radiological progression justifies biopsy of the tumour. In this case, an unusual tumour shows aggressive and invasive growth after surgical intervention and chemoradiotherapy.

There have only been 18 cases reported in the literature of an epidermoid tumor arising in the interhemispheric fissure. We report a 45-year-old woman with an epidermoid tumor arising in the interhemispheric fissure. She presented with a focal convulsive seizure. Radiological imaging identified a tumor in the interhemispheric fissure, which was subtotally removed due to adherence to both anterior cerebral arteries and the cerebral cortex. Epidermoid tumor was confirmed by histopathology. Epidermoid tumors rarely arise in the interhemispheric fissure, but are often adherent to surrounding structures and thus, as in the presented case, complete resection may be unwise.

We report a pediatric case of multiple sclerosis mimicking malignant brain tumor. A 11-year-old male presented with high fever, vomiting and partial convulsion of left lower extremity followed by left visual impairment. Magnetic resonance imaging (MRI) revealed a mass lesion in the right frontal lobe with significant peripheral edema. Incomplete ring-like enhancement by gadolinium administration was also noted. Open biopsy of the lesion was undertaken that provided pathological confirmation of demyelinating changes. Luxol fast blue stain and Bodian stain demonstrated extensive myelin loss and good preservation of axons. The patient was given intravenous high-dose methyl-prednisolone and sequential oral low-dose prednisone, which improved his neurological status progressively. The lesion was dramatically decreased in size and finally disappeared. Although many tumor-like demyelinating lesions were reported previously, pediatric cases of multiple sclerosis mimicking malignant brain tumor are extremely rare. Pathological examinations including Luxol fast blue stain and Bodian stain are essential for exact diagnosis.

Primary or secondary angiosarcoma in the central nervous system is rarely reported. We present a rare case of cerebral angiosarcoma, which comprised both poorly-differentiated solid areas and well-differentiated areas that led to the misdiagnosis of cavernous angioma. A 79-year old woman presented with an intracerebral hematoma in the left frontal lobe that was misdiagnosed as a hemorrhage from a cavernous angioma at initial operation. At a second surgery, the lesion was diagnosed as angiosarcoma involving the cerebellum, heart, femur, sacro-iliac bones and other locations. An autopsy suggested that the angiosarcoma of the heart was the primary lesion, which was occult at the time of the initial operation. Angiosarcoma may have areas with different degrees of differentiation and when a cavernous angioma is suspected histopathologically, the specimen should also be carefully explored for poorly-differentiated areas and the diagnosis of primary or secondary angiosarcoma considered.

Antisense strategy using synthetic oligodeoxynucleotides has been applied to the suppression of specific gene expression, the modulation of various gene expression, and its biological activity. Antisense strategy is applicable for the growth suppression of glioma cells. Several genes, including transforming growth factor-alpha, basic fibroblast growth factor, fibroblast growth factor receptor 1, vascular endothelial growth factor, telomerase, topoisomerase II alpha-subunit, protein kinase C-alpha, and microtubule-associated protein 1A, have been targeted by antisense strategy in glioma cells. These antisense strategies provide a potential novel antitumor therapy for gliomas.

Microtubule-associated proteins (MAPs) are essential for various cellular processes such as mitosis. The aim of this study was to verify that the suppression of MAP 1A using antisense oligonucleotide can suppress the in vivo proliferation of C6 glioma cells transplanted in rat brain. After 7 days from transplantation, antisense, sense or scramble phosphorothioate oligodeoxynucleotide for MAP 1A mRNA was gradually delivered into the established tumours through amini-osmotic pump. The mean diameters of the tumours from rats treated with antisense, sense and scramble phosphorothioate oligodeoxynucleotide for MAP 1A mRNA were 2.33, 4.625 and 5.25 mm. There was statistically significant difference in diameters of tumours between rats treated with antisense oligodeoxynucleotide, and those treated with sense or scramble oligodeoxynucleotide. This study strongly suggests the important role of MAP 1A in cell proliferation and its suppressionmay serve as a novel antitumour therapy for gliomas.

O(6)-methylguanine-DNA methyltransferase (MGMT) mRNA expressions were examined in 100 neuroepithelial tumors by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) using SYBR Green I. The mean relative quantitation value of MGMTmRNA normalized to the level of beta2-microglobulin for 100 tumors was 5.3 +/- 11.2. The mean value of 41 glioblastomas was significantly higher than that for the other 59 tumors (p = 0.0008 by Student's t-test). In contrast, the means of 19 low-grade gliomas and 12 medulloblastomas were significantly lower than that of other tumors (p = 0.0282 and p = 0.0456 by Student's t-test). Among the 55 retrospective patients who had been treated with 1-(4-amino-2-methyl-5-pyrimidynyl)methyl-3-(2-chloroethyl)- 3-nitrosourea hydrochloride (ACNU), the value was a significant independent predictor of the effect of initial therapy with ACNU (p = 0.0007 by Mann-Whitney U-test) and the survival period (p = 0.0175 by Wald test). The value >or=1 was the most significant factor in predicting the initial effect of treatment by multi-variant regression analysis (p < 0.0001). These results suggest that our individual adjuvant therapy based on MGMTmRNA expression may be improved by the application of real-time quantitative RT-PCR., (Copyright 2002 Wiley-Liss, Inc.)

To clarify the influence of biochemotherapy on the progression of astrocytic tumors, the expression of O6-methylguanine DNA-methyltransferase (MGMT) mRNA, as well as of other drug-resistance- and drug-sensitivity-related genes such as multidrug resistance gene 1, multidrug resistance-associated protein, glutathione S-transferase-pi, DNA topoisomerase II, and interferon receptor mRNA, and the interferon regulatory factor (IRF)-1 and -2 ratios in gliomas were investigated by quantitative reverse transcription-polymerase chain reaction (RT-PCR). The mean MGMT/beta2-microglobulin (beta2-MG) ratio for 130 neuroepithelial tumors was 8.2 +/- 17.8. The mean ratio of 45 glioblastomas was significantly higher than that for the other 85 tumors. In contrast, the mean of 26 low-grade gliomas was significantly lower than that of other tumors. The mean IRF-1/IRF-2 ratio of 16 other brain tumors that mainly consisted of medulloblastomas was significantly greater than that of the other 114 tumors. Almost no significant differences were observed between primary and recurrent tumors in the expression of any gene, and before and after therapy with corresponding drugs. The mean MGMT/beta2-MG ratio in primary glioblastomas was significantly higher than that in secondary tumors. These findings suggest that native drug resistance is more important than acquired resistance when glioma therapy is considered.

New adjuvant therapy individualized by the results of reverse transcription-polymerase chain reaction (RT-PCR) for drug-resistance genes has been used to treat malignant gliomas. Protocol studies for malignant gliomas were not so encouraging in their therapeutic results because of heterogeneity and the various drug-sensitivities of the tumors. Individualization of glioma therapy is recommended. Drug-resistance genes messenger ribonucleic acid (mRNA) expressions were investigated in drug-resistant human glioma cell lines derived from U87MG and 46 frozen samples of retrospectively examined neuroepithelial tumors (12 low grade neuroepithelial tumors, 16 Grade III gliomas, 11 glioblastomas, and 7 other malignant neuroepithelial tumors such as medulloblastomas and primitive neuroectodermal tumors) by RT-PCR with the specific primers for O6-methylguanine DNA methyltransferase (MGMT), multidrug-resistance gene 1 (MDR1), multidrug-resistance-associated protein (MRP), and glutathione-S-transferase-pi (GST-pi). Thirty-seven preliminary individual adjuvant therapies (IAT) based on RT-PCR results, mainly in MGMT expression, were performed on 30 consecutive patients with neuroepithelial tumors. In the retrospectively examined series, the initial response to 1-(4-amino-2-methyl-5-pyrimidynyl) methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride (ACNU) was correlated most significantly to the MGMT mRNA expression among 11 independent prognostic factors (p = 0.0037) in multivariate logistic regression analysis. In the preliminary IAT, 17 of 32 evaluable therapies had a partial or complete response (53.1% response rate). Our IAT based on RT-PCR seemed to be more effective than conventional therapies for malignant gliomas.

The difference in biological features between recurrent and nonrecurrent intracranial chordomas has not been studied. In this study, proliferative potentials of chordomas were studied with an immunohistochemical staining method, mainly using anti-Ki-67 antibody, MIB-1, which is known to be available for archival paraffin sections, together with immunohistochemical studies on the expression of cell cycle or apoptosis-related proteins, including p53, cyclin D1, and bcl-2 proteins. The correlation among MIB-1 staining indices, the immunoreactivities of these proteins, and clinical courses of intracranial chordomas were analyzed retrospectively, and the statistically significant correlation between MIB-1 staining index (SI) and recurrence has been clarified. The mean MIB-1 SI of recurrent tumors was 10.2%, being shown to be higher than that of nonrecurrent tumors (2.8%). The immunohistochemically positive staining of cell cycle-related protein, especially p53 and cyclin D1 proteins, correlated well with recurrence and high MIB-1 SI. In conclusion, both the examination of proliferative potentials of chordomas using MIB-1 SI and the study of the immunoreactivity of p53 and cyclin D1 proteins are important for their biological and histopathological analyses and the prediction of future recurrence.

A 5-year-old girl and a 2-year-old boy presented with recurrent cerebral primitive neuroectodermal tumor (PNET) and were treated with intravenous administration of human fibroblast interferon (HuIFN-beta) which was continued as outpatient maintenance therapy. Both patients showed a partial response and were still alive 14 months and 2 years after diagnosis. Our results suggest that HuIFN-beta is an effective therapy for PNET, and can be used as long-term maintenance therapy without serious side effects.

In radiation therapy for malignant brain tumours, the dose of radiation that can be safely delivered to a tumour is limited by the radiation tolerance of the adjacent normal brain tissue. Among various radiation modalities to produce local tumour eradication without unacceptable complications, we chose a large, single irradiation dose during the operation (intra-operative radiation therapy, IORT). In contrast to X-ray or Cobalt-60 gamma ray irradiation, IORT with a high-energy electron beam delivered by the Shimadzu 20 MeV betatron provides acceptable dose homogeneity with rapid fall-off of the radiation dose beyond the treatment volume. Thus, IORT has the advantage of precise demarcation of the target volume, minimum damage to surrounding normal tissues, and a high absorbed target dose (15-25 Gy in 5-10 min). On the basis of our experience with 170 patients treated by IORT, we established the treatment indications and method in patients with malignant brain tumours. IORT with a dose of 15-25 Gy was delivered to widely resected tumours followed by external radiation therapy. No acute or subacute complications were observed. Treatment results of 30 patients with glioblastoma treated by IORT (mean 18.3 Gy) combined with external radiation therapy (mean 58.5 Gy) resulted in a median survival of 119 weeks and a 2-year survival rate of 61%.

We analyzed our treatment results of 71 operated patients with cerebral glioblastoma treated by conventional external radiation therapy (mean dose 60.2 Gy) combined with radiosensitizing agents. More than 50% reduction of tumor volume was obtained in 20 patients (28.2%). A response rate of at least 40% was obtained in patients treated with combined ACNU-vincristine-nicardipine, ACNU-5FU-hydroxyurea, or cisplatin alone. The combination of ACNU and vincristine with or without nicardipine resulted in significantly longer survival. The median survival in this group was 101.1 weeks and the two-year survival rate was 45.9%; these results were significantly better than those achieved with other ACNU combinations or other combinations without ACNU. In the analysis of survival, factors correlated to longer survival were a patient age of younger than 45 years, wide resection of the tumor, a good postoperative performance status (KS > or = 70%), a radiation dose of 68-72 Gy, small postoperative tumor remnants (< 20 cm3), no visible tumor after radiation therapy, and the administration of adjuvant chemotherapy. Maximum resection of the tumor and localized irradiation with a dose of 70 Gy combined with ACNU and vincristine appears to be the most effective treatment at present.

The purpose of this study is to quantify the relative contribution of the mechanisms in the absorption of edema fluid. The convection/diffusion and the comprehensive bulk flow model were applied for the finite element analysis of peritumoral brain edema. For clinical analysis, 90 meningiomas studied by MRI were selected. Serial CT scan and MRI were performed at 0, 2, 4, 6 hours after injection of Iopamidol or Gadpenteic acid respectively. Then the tracer distributions in the edematous brain was analyzed. The tracer movement in the brain is well represented by the convection/diffusion equation. The absence of the preferential fluid flow directing toward the ventricle indicates that a limited role of CSF sink action into the ventricle. From capillary surface area (240 cm2/g brain), capillary hydraulic conductivity (1.8 x 10(-8) ml/cmH2O/cm2/min) and the simulated average tissue pressure of 9.8 mmHg, maximum absorption rate into capillaries was estimated to be 0.003 ml/h/cm3 brain tissue. Considering the limited role of edema fluid clearance into the ventricle, the results indicate a possible role of subarachnoid CSF space for the clearance of edema fluid. The clearance of edema fluid into subarachnoid CSF space should be studied quantitatively. Finally, unification of the convection/diffusion and the comprehensive bulk flow model will provide a more quantitative analysis of edema formation and resolution by using MRI and tracer studies.

In order to elucidate the role of inflammatory cytokines in the central nervous system, we examined the production of two leukocyte chemoattractants, IL-8 and monocyte chemotactic and activating factor (MCAF) in brain tumor cell lines. The glioma cell lines tested exhibited high levels of IL-8 and MCAF mRNA expression upon stimulation with IL-1 or TNF-alpha, while none of the neuroblastoma cell lines expressed these cytokine mRNA. Both IL-8 and MCAF mRNA expression depended on the dose of IL-1 alpha and TNF-alpha and appeared very rapidly, reaching maximal levels at 3-6 hr, with substantial production of these cytokines in the culture supernatants. When various immunosuppressive drugs were tested, glucocorticoids but not other immunosuppressive drugs markedly inhibited the IL-1 or TNF-alpha-induced IL-8 and MCAF mRNA accumulation, suggesting that glucocorticoid is a potent regulator of these inflammatory cytokine production in the neural tissues. In addition, reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of IL-8 and MCAF mRNA expression in resected brain tumor tissues including glioblastoma, astrocytoma grade 2, ependymoma and medulloblastoma, indicating that these inflammatory cytokines are expressed in vivo.

Gliomatosis cerebri (GC) is a rare clinical entity characterized by diffuse and infiltrative overgrowth of the tumor cells. Most of the previously reported cases of GC were autopsy cases because the clinical diagnosis of GC has been difficult. The authors report four cases diagnosed clinically as GC. Cases 1 and 2 are females aged 19 and 69. Cases 3 and 4 are males aged 47 and 50. In the first three cases, CT findings were almost normal. MRI study, especially on its T2 weighted image (T2W1), clearly demonstrated the wide extent of the infiltration of the tumor cells along the white matter. The last case occurred in the pre-MRI era, but contrast enhanced CT showed a bilateral periventricular high density area accompanied by diffuse low density white matter. Three of them underwent echo-guided needle biopsy, and one underwent partial excision of the lesion. Histological diagnosis was glioblastoma in Cases 1 and 4, and anaplastic astrocytoma in Cases 2 and 3. Difficulty in the clinical diagnosis of GC has been based on the fact that traditional imaging studies, including CT, can not clearly show the extent of tumor cell infiltration. MRI study is a very sensitive imaging technique which can easily demonstrate the area infiltrated by glioma cells. So we may be able to make clinical diagnosis of GC, coupling the data from MRI study and brain biopsy. The authors expect that accumulation of clinical experiences of GC may give useful information for the investigation of "invasion", which is one of the major problems in the treatment of malignant gliomas.

We investigated the correlation between the technique of radiation therapy used and the regrowth pattern of recurrent glioblastoma multiformes of 48 patients who received initial radiation therapy in our clinic from April, 1974 to March, 1988. Three different techniques have been applied to patients with brain tumors: whole brain irradiation, generous local irradiation (the parallel opposing technique or three field technique, in which the treatment field fully covers the peritumoral low density area on CT images), and restricted local irradiation (the rotation technique, in which the treatment field is restricted to within about 2 cm of the tumor margin on CT images). Radiation dose of the treatment field was over 45 Gy in every case. The regrowth pattern was defined as being one of the followings: inside of the treatment field, outside of the treatment field (this includes the boundary zone between the treatment and the non-treatment field), and remote from the treatment field-which mostly due to the tumor spreading through ventricular systems. In all 7 cases that received whole brain irradiation, tumor recurred inside of the treatment field. Two of these 7 cases showed remote recurrence at the same time. Mean duration time to recurrence was 36.3 weeks. In 27 (90%) of 30 cases that received the generous local irradiation, tumor recurred within the treatment field. Only one showed outside recurrence, and two other cases showed remote recurrence. The mean duration time to recurrence in this group was 32.2 weeks. Of 11 cases that received restricted local irradiation, 3 cases (27%) showed recurrence outside of the treatment field, one showed recurrence in a remote area. The mean duration time to recurrence in this group was 38.3 weeks, but was not significantly longer than that of other two groups. These results indicate that restricted local irradiation sometimes fails to cover the tumor invaded area, and that the results of treatment using generous local irradiation are almost the same as those using whole brain irradiation. This suggests that generous local irradiation is advantageous to patients with brain tumors since the incidence of subacute side effects such as mental deterioration is much lower in local irradiation as compared with whole brain irradiation.

Perturbed cell kinetics in 9 L rat brain tumor treated with dibutyryl cyclic AMP (DBcAMP) and theophylline were studied. The methods include a procedure for simultaneous flow cytometric measurements of cellular DNA and amount of bromodeoxyuridine (BrdU) incorporated into cellular DNA and a procedure for flow cytometric measurements of cellular DNA and RNA. Propidium iodide and monoclonal antibody against BrdU were used as a fluorescent probe for total cellular DNA and for BrdU incorporated into cellular DNA, respectively. For analysis of cellular DNA and RNA, acridine orange staining was also applied. The results were as following; cells in S and G2 phase at the time of drug administration (1 mM, each) can undergo mitosis even though a considerable prolongation of G2 phase was apparent. However, cells in G1 at the time of drug administration were arrested in that phase, whereas those cells in S or G2 were able to complete one mitosis before becoming arrest in the G1 phase. These results have been inferred from the study by autographic method with 3H-thymidine. However, using these new methods, these were clearly demonstrated and furthermore, flow cytometric analysis with acridine orange revealed that DBcAMP induced 9 L cells into resting position (G0 cells).

Twelve patients with ependymomas received a 30- to 60-minute intravenous infusion of bromodeoxyuridine (BrdU), 150 to 200 mg/m2 at surgery, to label tumor cells in the DNA synthesis phase. Labeled cells were detected in excised tumor specimens by indirect immunoperoxidase staining using anti-BrdU monoclonal antibody as the first antibody. The BrdU labeling index (LI, defined as the percentage of labeled cells in relation to the total number of cells scored) was calculated for each specimen. All four spinal cord ependymomas had a BrdU LI of less than 1%, which is consistent with our clinical experience that most such tumors grow slowly and have an excellent prognosis. Five of the eight intracranial ependymomas also had a low BrdU LI of approximately 1% or less, and three had a BrdU LI of 3.2%, 3.4%, and 4.8%. The latter three tumors, only one of which was diagnosed as a malignant ependymoma at the time of study, were either recurrent or recurred within 2 years after gross or subtotal removal. Cytologic analysis of cerebrospinal fluid (CSF) was performed in five cases; CSF seeding of tumor cells was found in only one patient, who had a malignant ependymoma. A high BrdU LI did not always correlate with CSF seeding. Measurement of the LI using BrdU and anti-BrdU monoclonal antibodies can provide more accurate information on the proliferative potential of individual tumors and may lead to a more rational grading system of ependymomas. The results of such studies do not always predict the potential for CSF seeding.

A case of primary cerebral malignant lymphoma associated with hydrocephalus is reported. The patient was a 54 year-old male who enjoyed good health until the onset of headache and vomiting 4 weeks before admission. His consciousness was alert and neurological examination revealed severe papilloedema with retinal hemorrhage. No lymph node or abdominal tumor enlargement were noted. CT scan and MR images revealed no abnormal lesion except mild ventriculomegaly. CSF study revealed mild elevation of protein and sugar and cell count was 66/3. CSF cytology revealed atypical lymphoid cell with irregular nuclear contour and large nucleolus. Immunological marker studies of the tumor cell revealed increasing of anti J-5 (CD10), anti B-4 (CD19) and OKT-IA1. The patient was treated by a whole brain irradiation and chemotherapy after V-P shunt. It is 12 months since the operation, and the patient's condition is still good.

Five patients with various brain tumors received bromodeoxyuridine (BrdU), 150-200 mg/m2 i.v., at the time of craniotomy. Biopsied materials were fixed in 70% ethanol, sectioned, denatured with hydrochloric acid, and reacted with monoclonal antibodies against BrdU. Immunofluorescence and immunocytochemical methods were used to visualize BrdU-labeled nuclei. Our results showed that both methods demonstrated BrdU-labeled nuclei satisfactorily in tissue sections. Thus, BrdU can be used to measure the proliferative potential of human tumors in situ.

We studied the effects of increased-dose radiation therapy in terms of survival time and improvement in the quality of life in 50 glioblastoma patients with minimal residual tumors. 1) Intraoperative radiotherapy ( IOR , 1,000-2,000 rad) was applied in 13 cases; the 2-year survival was 41.6%. However, in 9 patients who had undergone macroscopic total removal the 2-year survival rate was 68.6%. 2) Wide resection with necrotomy after conventional external irradiation was used in 9 cases and conformation irradiation was added; their 2-year survival was 44.4%. 3) In 22 patients who received only conventional irradiation following surgery, the 2-year survival was 7.5%.

Thirty-eight patients undergoing surgical removal of neuroectodermal tumors of the central nervous system were given a 1-hour intravenous infusion of bromodeoxyuridine (BUdR), 150 to 200 mg/sq m, to label tumor cells in the deoxyribonucleic acid (DNA) synthesis phase (S-phase). The excised tumor specimens were divided into two portions: one was fixed with 70% ethanol and embedded in paraffin and the other was digested with an enzyme cocktail to make a single-cell suspension. The paraffin-embedded tissues were stained by an indirect peroxidase method using anti-BUdR monoclonal antibody (MA) as the first antibody. Single-cell suspensions were reacted with fluorescein isothiocyanate (FITC)-conjugated anti-BUdR MA's for flow cytometric analysis. S-phase cells that had incorporated BUdR into their DNA were well stained by both methods. The percentage of BUdR-labeled cells, or S-phase fraction, was calculated in tissue sections by microscopic examination and in single-cell suspensions by flow cytometric analysis. The biological malignancy of the tumors was reflected in the S-phase fractions, which were 5% to 20% for glioblastoma multiforme, medulloblastoma, and highly anaplastic astrocytoma, but less than 1% in most moderately anaplastic astrocytomas, ependymomas, and mixed gliomas. Two juvenile pilocytic astrocytomas and two low-grade astrocytomas from children had high S-phase fraction despite the fairly benign and slow-growing nature of these tumors. These results indicate that the S-phase fraction obtained immunocytochemically with anti-BUdR MA's may provide useful information in estimating the biological malignancy of human central nervous system tumors in situ.

FITC-conjugated anti-bromodeoxyuridine (BrdUrd) monoclonal antibody (anti-BU-MAb) was used to detect S-phase cells of 9L rat brain tumor cells in vitro and in situ. Monolayer 9L cells were treated with 0.625-20 microM of BrdUrd for 30 min, harvested, and reacted with a 1:100 dilution of FITC-conjugated anti-BU-MAb and analyzed with a flow cytometer. BrdUrd-treated cells stained satisfactorily with antibody. Values obtained for the labeling index using this method (48.6%) were 10%-20% higher than the fraction of cells in S-phase calculated from DNA histograms or as the labeling index calculated from autoradiographs of cells pulse-labeled with 3H-thymidine. BrdUrd (1-40 mg/kg) was administered by i.p. injection to rats bearing 9L brain tumors. Single cell suspensions obtained by disaggregation of excised tumors were stained with anti-BU-MAb. The percentage of fluorescent cells (15.9%) calculated using this method was similar to that of S-phase cells (17.2%) calculated from DNA histograms and from autoradiographics for tumor bearing rats pulse-labeled with 3H-thymidine in situ. The antibody staining technique is a rapid and accurate method for various cell kinetic studies both in vitro and in vivo in a rat model, and has promise as a technique for the study of cell kinetics in humans.

Cell kinetic studies on various human brain tumors were reviewed. Most studies have been carried out by means of 3H-thymidine and autoradiography in the past two decades. The average labeling index (LI) obtained from a pulse of 3H-thymidine is very high in medulloblastomas and glioblastoma multiforme (5-15%), low in well-differentiated gliomas (less than 1%), and intermediate in anaplastic astrocytomas. The higher the LI, the faster the tumor grows, probably reflecting a larger growth fraction. Therefore, measurement of the LI appears to be very helpful in predicting the prognosis of the patient as well as potentially helpful in the design of chemotherapy. However, isotopic studies not only take a long time to complete, but also have severe limitations because of potential radiation hazard to patients and environmental pollution. Development of an anti-BUdR (or BrdUrd) monoclonal antibody to detect nuclei which incorporate BUdR is a breakthrough that can expand this line of research, since: 1) BUdR is non-radioactive and doses needed for this purpose are virtually non-toxic; 2) use of flow cytometric analysis of FITC conjugated anti-BUdR antibody facilitates rapid acquisition and data analysis and, 3) it will be technically easier to study the proliferative capacity of patients with brain tumors. A 30 min exposure of 9L monolayer cells to 10 X 2-10 X 2(-4) microM BUdR produced satisfactory results against a 1: 60 dilution of FITC conjugated antibody (Becton-Dickinson, Mt. View, CA). Also 9L cells which were exposed various concentrations of BUdR (10 X 2-10 X 2(-4) microM) for 30 min were harvested in single cell suspension and reacted with the antibody and analysed with flow cytometry. Fluorescent nuclei (48.6%) were similar to the fraction of cells in S phase analysed from DNA histograms as well as the LI obtained from autoradiographic study after a pulse of 3H-thymidine. After 1-40 mg/kg of BUdR were injected into the peritoneal cavity of rats with 9L brain tumors, tumors were removed 1 hr later and digested with enzyme cocktail, fixed with 70% ETOH, denatured with HCl, and stained with FITC conjugated anti-BUdR antibody. The nuclei which reacted with the antibody were discriminated well with flow system at 488 nm light through LP 515 and SP 560 filters. 10.0-15.5% of the cells were fluorescent in each group and the intensity of fluorescence was dose dependent although not strictly proportional to the amount of BUdR administered.(ABSTRACT TRUNCATED AT 400 WORDS)

At the time of surgery, 18 patients with various brain tumors were given a 1-h i.v. infusion of bromodeoxyuridine (BrdUrd), 150-200 mg/m2. At an infusion rate of 200 mg/m2/h, serum BrdUrd levels of 8 microM were achieved. After the infusion, tumor tissue was obtained and divided into two portions. One portion was fixed in 70% ethanol, embedded in paraffin, and sectioned; the sections were deparaffinized, denatured with 2 N HCl, and reacted with monoclonal antibodies against BrdUrd (anti-BrdUrd MAb). BrdUrd-labeled nuclei were demonstrated satisfactorily by an indirect peroxidase method. The other portion was dissociated into single cells with a DNase enzyme cocktail and reacted with FITC-conjugated anti-BrdUrd MAb to determine the percentage of BrdUrd-labeled cells or with chromomycin A3 for DNA analysis. The single-cell suspensions were analyzed by flow cytometry. The fraction of S-phase cells in the tissue sections was similar to both the percentage of BrdUrd-labeled nuclei and the S-phase fraction determined by flow cytometric analysis. The results obtained with BrdUrd-labeled nuclei were similar to those obtained from previous autoradiographic studies of various brain tumors exposed to a pulse of 3H-thymidine. Since BrdUrd is not radioactive and is nontoxic at the dosage used, these techniques, together with the histopathological diagnosis, may help to predict the biological malignancy of individual tumors.

It is still controversial whether edema fluid around a malignant brain tumor is derived from capillaries inside the tumor itself or not only from tumor vessels but also from peritumoral tissue. The purpose of this study is to clarify the region where the capillary permeability is increased using double autoradiographic method. A suspension of 1 X 10(4) rat glioma cells (RG-12) was stereotactically implanted into the right basal ganglia of C-D Fisher rats. In this model, all animals are dead 20 +/- 1 days after the implantation. Two kinds of tracers, 14C-alpha-aminoisobutyric acid (AIB) and 131I-human serum albumin (HSA), were administered 14 to 17 days after the tumor implantation. In all rats, 14C-AIB was intravenously injected 10 min before decapitation. 131I-HSA was given 10 min before decapitation in five animals (group 1), one hour before in four animals (group 2), and six hours before in three animals (group 3). Autoradiograms for 131I-HSA were exposed for initial one to two days after the decapitation, while the exposure for 14C-AIB was delayed until 4 months later. Autoradiograms of these two tracers were compared with the corresponding sections with H-E stain for neuropathological examination. In group 1, the distribution of HSA as well as AIB was quite similar to tumor itself. In group 2 and group 3, the distributions of AIB were similar to the tumor. However, the distributions of HSA were 1.4 fold larger (group 2) and 3.6 fold larger (group 3) than the tumor and expanded into the peritumoral region.(ABSTRACT TRUNCATED AT 250 WORDS)

Bromodeoxyuridine (BUdR) is a radiosensitizer that can be incorporated into cellular DNA as a substitute for thymidine at the time of DNA synthesis. As the steady-state arterial concentration of BUdR given by means of intravenous infusion was recently presented, the possibility of revival of BUdR as a radiosensitizer administered by the intravenous route was suggested. Based on the experience of BAR therapy and phase-I studies by NIH and UCSF, 12 hours of BUdR at a dose of 800-1,000 mg/m2 for five days a week was given to 23 patients with primary and secondary malignant brain tumors during radiation therapy. Radiation therapy was planned at a weekly dose of 10 Gy for five to six weeks. Fifteen patients received 1,000 mg/m2 of BUdR; six of them tolerated more than three weeks of treatment. In eight patients given doses of 800 mg/m2, five patients tolerated more than three weeks. The most remarkable toxic effects were myelosuppression and stomatitis, which were major obstacles to maintaining the schedule. More than 50% reduction of tumor volume was obtained in five of 12 cases of evaluated gliomas (42%) and three of four cases of metastatic tumors (75%). The median time to tumor progression in seven patients with glioblastoma was 37 weeks.

One hundred forty-three patients with gliomas of astrocytic origin (61 with glioblastomas multiforme (GM) and 82 with astrocytomas) received an intravenous infusion of bromodeoxyuridine (BUdR), 150-200 mg/m2 at the time of surgery, to label tumor cells undergoing DNA synthesis. BUdR-labeled cells were identified by the indirect immunoperoxidase method using anti-BUdR monoclonal antibodies. The percentage of BUdR-labeled cells, or BUdR labeling index (LI), was calculated by microscopic examination of selected viable areas of the tissue sections. The GMs had a median LI of 7.5%, and three quarters of these tumors had LIs greater than 5%. Highly anaplastic astrocytomas (HAAs) and moderately anaplastic astrocytomas (MoAAs) had median LIs of 2.3% and less than 1%, respectively. Among the HAAs, the LI was 1% to 5% in 56% of tumors, greater than 5% in 26%, and less than 1% in 18%. More than 60% of MoAAs had LIs less than 1%, which agrees with the slow clinical progression of this type of tumor, and the remainder had LIs of 1.4% to 9.3%. These results show that histopathologically similar tumors may have different proliferative potentials. Measuring the proliferative potential of individual gliomas is therefore crucial for predicting the prognosis more accurately and for devising more tumor-specific treatment regimens for individual patients.

Bromodeoxyuridine (BrdU), 150 to 200 mg/m2, was administered at the time of operation to 20 pediatric patients with neuroectodermal tumors to label tumor cells in the S phase. Immunocytochemical techniques were used on excised tumor specimens to detect cells containing BrdU, and the BrdU labeling index (LI) was calculated as the number of BrdU-labeled cells divided by the total number of cells counted. Four medulloblastomas, three glioblastomas multiforme, and two highly anaplastic astrocytomas had average BrdU LIs of 13.0 +/- 3.0% (SE), 12.7 +/- 4.3%, and 14.6 +/- 6.7%, respectively. Three of nine moderately anaplastic astrocytomas had BrdU LIs of greater than 1% (average, 6.5 +/- 2.4%), whereas six had LIs of less than 1%. In two juvenile pilocytic astrocytomas, which are considered slow-growing, the BrdU LIs were unexpectedly high, averaging 6.5 +/- 1.4%. Thus pediatric medulloblastomas, glioblastomas multiforme, highly anaplastic astrocytomas, and a minority of moderately anaplastic astrocytomas had high proliferative potentials, whereas most of the moderately anaplastic astrocytomas had low proliferative potentials. Although the number of cases in this study is still too small to yield statistically significant comparisons, the results indicate that some pediatric tumors have considerably higher LIs than histologically similar adult tumors studied previously.

Regional blood flow and capillary permeability in the experimental brain tumors and their surrounding brain tissue of rats were measured with quantitative 14C-antipyrine and 14C-alpha-aminoisobutyric acid (AIB) autoradiographic method. The pharmacokinetic implications with respect to drug delivery to tumor tissue and the effect of ionizing irradiation were discussed in these physiological measurement. A suspension of 1 X 10(4) rat glioma cells (E239 RG 12) was stereotactically implanted into the right basal ganglia of CD-Fisher rats, and spherical brain tumors developed 10-17 days after implantation with a diameter of 1--5 mm. Autoradiographic investigations were performed for rats with small tumors (1--2 mm in diameter) and large tumors (4--5 mm in diameter). The uniform blood flow (91.7 +/- 13.1 ml/100 g/min: mean +/- S.E.) was observed in small tumors with the patchy low flow area (56.7 +/- 12.5 ml/100 g/min) surrounding the tumor. In large tumors, the blood flow was markedly decreased in the central part of the tumor (28.3 +/- 2.4 ml/100 g/min) with a ring shaped high flow area in the peripheral part (59.3 +/- 5.9 ml/100 mg/min). The blood flow in the brain adjacent to the tumor (30.5 +/- 2.5 ml/100 g/min) was lower than that in the peripheral part of the tumor. The uptake of 14C-AIB was quite similar to that of 14C-antipyrine suggesting the smaller permeability in the central part of the tumor. Neuropathological studies did not reveal necrotic foci, but viable cells in these areas.(ABSTRACT TRUNCATED AT 250 WORDS)

Sixteen patients with glioblastoma multiforme received a 1-h intravenous infusion of 5-bromo-deoxyuridine (BrdU), 150-200 mg/m2 at the start of surgery, to label S-phase cells in tumor tissue. Labeled cells of vascular components and of tumor parenchyma were detected in excised tumor specimens by indirect immunoperoxidase staining using anti-BrdU monoclonal antibodies followed by periodic acid-Schiff staining. The BrdU labeling index (LI, defined as the percentage of labeled cells in relation to the total number of cells scored) was calculated separately for vascular components and tumor parenchyma in each specimen. The BrdU LI of vascular components of glioblastoma multiforme was remarkably higher than that of normal brain (1.1%-8.7% vs. less than 0.05%). The mean BrdU LIs of vascular components and tumor cells in eight primary glioblastomas were 4.5 +/- 0.8% (mean +/- SE) and 9.9 +/- 1.1%, respectively, while the corresponding BrdU LIs in eight recurrent tumors were 2.7 +/- 0.5% and 9.3 +/- 0.7%. The differences in the BrdU LIs of primary and recurrent tumors were not statistically significant, but the BrdU LI of vascular components was consistently much lower than that of tumor cells. BrdU labeling of vascular components was inconsistent and occurred mostly in glomerular-shaped vessels, but only about 20% of them contained labeled cells. These results suggest that unusual vascular proliferation, such as the formation of glomerular-shaped vessels and endothelial or adventitial proliferation, in glioblastoma multiforme may have been programmed to slow down or even to cease at a certain stage, and is not likely to be the result of neoplastic transformation.

Sixteen patients with brain tumors were given a 30- to 60-minute intravenous infusion of bromodeoxyuridine (BUdR), 200 mg/sq m. Grossly viable fragments were taken from the biopsied tumor specimens and divided into two portions. One portion was dissociated into single cells, stained both with fluorescein isothiocyanate (FITC) using anti-BUdR monoclonal antibody as the first antibody and with propidium iodide (for deoxyribonucleic acid), and analyzed by flow cytometry (FCM). The labeling index (LI) was calculated as the number of FITC-labeled cells expressed as a percentage of the total number of cells analyzed. The other portion was fixed in 70% ethanol, embedded in paraffin, sectioned, and stained with immunoperoxidase using anti-BUdR monoclonal antibody as the first antibody. The LI of these tissue sections was calculated in two ways: from selected areas in which the labeled cells were evenly distributed and from the entire tissue section. The LI's obtained by FCM correlated closely with those from the entire tissue sections (r = 0.99, p less than 0.000001) and were usually lower than LI's from selected areas of tissue sections. The LI's determined by FCM also correlated well with the LI's from selected areas of tissue sections (r = 0.82, p less than 0.00012), despite the difference in values between them. Thus, the FCM-derived LI and the tissue LI can both provide useful information for predicting the biological malignancy of individual tumors and for designing treatment regimens for individual patients with brain tumors; however, different standards should be used to interpret the LI's obtained by these two methods.

The halogenated pyrimidine analogue bromodeoxyuridine (BrdU), which is incorporated into nuclei during DNA synthesis, has long been known to be a radiation sensitizer. Since 1965, BAR therapy (BrdU-antimetabolite-radiation therapy), in which BrdU was administered intraarterially as a radiosensitizer, has been applied to patients with malignant gliomas and the improvement in survival rate within two years has been reported. Recently, intravenous infusion of BrdU has prove to be sufficiently effective as a radiosensitizer and BrdU is still being utilized as a chemical modifier for patients with malignant gliomas. Misonidazole was developed as a hypoxic cell sensitizer and was expected to enhance the radiation response of malignant tumors. However, the clinical trial of misonidazole in patients with brain tumors showed little clinical benefit of this agent as a radiosensitizer, and therefore it is no longer used in the treatment of malignant gliomas. Synchronized chemoradiotherapy, in which alkaloid and alkyl agents are used to accumulate cells into the radiosensitive G2 and M phases, was developed for the treatment of malignant gliomas in 1976 and significant improvement in survival has been reported. However, phase II studies demonstrated that radiotherapy with alkyl agents such as BCNU and ACNU did not prolong the survival of patients with malignant gliomas as compared with radiotherapy alone, although they did increase the response rate. Since 1985, the Brain Tumor Interferon Study Group has clinically applied one of the biological response modifiers (BRM), interferon-beta (INF-beta) as a chemical modifier in patient with malignant gliomas. They have reported that the response rates in patients treated with ACNU + radiation and INF-beta + ACNU + radiation were 19.6% and 41.2%, respectively. Their results suggested that IFN-beta with ACNU was a promising regimen as a chemical modifier in radiotherapy for patients with malignant gliomas. In order to improve the rate of local control of malignant gliomas and to prolong the survival of patients, it is necessary to continue to seek effective chemical modifiers including BRMs, as well as to develop irradiation techniques.

A man aged 31 with remittent fever, leukopenia, polyuria, and splenomegaly in addition to chronic and recurrent episodes of neurologic abnormalities was examined pathologically. A chronic type of perivenous encephalomyelitis was found in the CNS, and chronic inflammatory lesions were noticed in the infundibulo-hypophyseal system and also in the peripheral nerves. Moreover, hemophagocytic lymphohistiocytosis predominated in the reticuloendothelial system. This seemed to be the first adult case of demyelinating disease with hemophagocytic lymphohistiocytosis.

One hundred fifty-four patients with brain tumors of various types were given an intravenous infusion of the thymidine analogue bromodeoxyuridine (BUdR), 200 mg/m2, at the time of surgery but before biopsy of the tumor to label S-phase cells. Excised tumor specimens were fixed, sectioned, and stained by immunoperoxidase methods to detect BUdR. The labelling index (LI), or percentage of BUdR-labelled cells, was calculated for each tumor specimen. The LIs of glioblastomas, medulloblastomas, and most highly anaplastic astrocytomas were 5% to 20%. The majority of moderately anaplastic astrocytomas showed LIs of less than 1%, but 30% of them had LIs similar to those of highly malignant gliomas. Most pituitary adenomas and neurinomas showed LIs of less than 1%. Nonmalignant meningiomas had LIs of less than 1%, whereas malignant meningiomas had LIs higher than 2.7%. This is an important observation, because malignant meningiomas are not well-defined histopathologically and their growth rate and rate of recurrence cannot be predicted by current diagnostic procedures. By estimating the proliferative potential of individual tumors more precisely, the BUdR LI supplements histopathological diagnosis, allowing a more accurate estimate of prognosis and facilitating the design of treatment regimens for individual patients.